russell_apr_20110102

End-to-End Single-Use Process for Monoclonal

Antibody Manufacturing

� e Adaptable Laboratory

Raman Spectroscopy Applied to Pharmaceutical Tablets

Amorphous Solid Dispersions

Approaches to Quanti� cation by Powder X-ray Di� ractionw

ww.a

mer

ican

phar

mac

eutic

alre

view

.com

American Pharmaceutical

The Review of American Pharmaceutical Business & Technology

Volume 14 Issue 1 | Jan/Feb 2011

American Pharm

aceutical Review

| Volume 14 Issue 1 | Jan/Feb 2011

SHOW ISSUE

FC_APRJanFeb2011.indd 1 2/11/11 3:53 PM

endo ad campaign 20102.indd 1 10/15/2010 12:10:01 PM

Xcelolab™ Powder DispenserProvides fast, fl exible, precision powder dispensing for any laboratory. Performs accurate, closed loop, weight dispensing that is diffi cult and potentially inaccurate by hand or by other laboratory methods.

Xcelodose® S Precision Powder Micro-Dosing SystemPrecisely dispenses drug substances alone, as low as 100µg, into capsules without excipients or bulking agents. This allows companies to eliminate costly and time-consuming compatibility, stability and preformulation studies during early stage drug development.

CFS 1200 and the NEW CFS 1500 C Liquid Filling & Sealing MachinesThese fully automatic cGMP compliant machines fi ll and seal liquids/semi-solids into two-piece hard capsules at speeds of 1,200 and 1,500 capsules per hour. This helps scientists exploit the potential of lipid-based formulations for poorly soluble compounds.

CapsuleCaddy™ ContainerThe perfect complement to our R&D machines, it contains 10,000 Licaps®, DBcaps®, Vcaps® Plus or Coni-Snap® capsules.

To receive a full R&D portfolio, call 888-783-6361 or send an email to marketing.AMER@pfi zer.com.

www.capsugel.com

Achieving Faster Time to First in Human™

NEW

NEW

APR_19818_FIN_7-26-10.indd 1 7/26/10 11:37 AM

EditorialEDITOR | Emily Johnson

[email protected]

SalesACCOUNT MANAGER | Southeast | Joel Kern

[email protected] SALES MANAGER | Gary King

[email protected] SALES MANAGER | Laura Zoibi

[email protected]

CorporatePRESIDENT | Nigel Russell

CIRCULATION DIRECTOR | Steve Koppelman

ProductionART DIRECTOR | Jennifer Campbell

PRODUCTION MANAGER | Brandon WaggonerOFFICE MANAGER | Sandra Anwarzai

American Pharmaceutical ReviewPublished by

Russell Publishing L.L.C.9225 Priority Way West Drive, Suite 120

Indianapolis, IN 46240-1467 USA

Tel: 317-816-8787 | Fax: 317-816-8788www.russpub.com

E-mail: [email protected]

Website: www.americanpharmaceuticalreview.com

Printed in the USA

American Pharmaceutical Review (ISSN - 1099-8012) is pub-lished seven times a year by Russell Publishing LLC, 9225 Priority Way West Drive, Suite 120, Indianapolis, Indiana 46240-1467. Subscription Rate (7 issues) 1 year $135.00. Periodicals postage paid at Indianapolis, IN 46206 and additional mailing offices.

POSTMASTER: Send change of addresses to Russell Publishing LLC, 9225 Priority Way West Drive, Suite 120, Indianapolis, Indiana 46240-1467. Publication Agreement #40739004. Return Undeliverable Canadian Addresses to: Station A, PO Box 54, Windsor ON N9A 6J5, E-mail: [email protected]

Copyright rests with the publishers. All rights reserved ©2010Russell Publishing L.L.C.

No part of this publication may be reproduced, stored in a retrieval system or transmitted in any form or by any means electronic, mechanical, photocopying, recording or other-wise without the prior written permission of the copyright holder. While the publishers believe that all information contained in this publication was correct at the time of going to press, they can accept no liability for any inaccuracies that may appear or loss suffered directly or indirectly by any reader as a result of any advertisement, editorial, photo-graphs or other material published in American Pharmaceuti-cal Review.

2 | | January/February 2011

Dependable and RelevantYour Source for Information in the

Pharmaceutical Industry

American Pharmaceutical Review, Pharmaceutical Outsourcing, and

International Drug Discovery bring the most up to date reviews, news, trends and comments

to an ever-changing industry. With their complementary websites, e-newsletters and webcasts they offer the complete spectrum of marketing avenues for any company that

operates in the life science arena.

Amerian Pharmaceutical Review, Pharmaceutical Outsourcing, and International Drug Discovery are products of Russell Publishing

9225 Priority Way West Drive, Indianapolis, IN 46240 317-816-8787 | Fax 317-816-8788

Harnessing the Power of Knowledge to Drive World Class Science and Technology

April 11-15, 2011JW Marriott San antonio Hill Country | San antonio, texaS

www.pda.org/annual2011

The Parenteral Drug Association presents...

1946

2011

65TH ANNIVERSARY

Meeting April 11-13 | exhibition & Career Fair April 11-12

PDa/FDa ProCess ValiDation guiDanCe Post ConFerenCe WorkshoP April 13-14 | Courses April 14-15

www.pda.org/annual2011

The Opening Keynote addresses include:

Join more than 50 speakers in over 45 presentations in sessions that will discuss:

Quality Science

Manufacturing/Process Science

Development Science

Outsourcing/Supply Chain

New in 2011 is the Fundamentals Track – This track is designed for those who are new to the pharmaceutical/biopharmaceutical industry or have recently changed jobs with a change in focus.

Additional Educational and Training Opportunities!

Attend the PDA Process Validation Guidance Post Conference Workshop to learn about US and international regulations, technology transfer, documentation strategies, post approval reporting and more!

PDA’s Training and Research Institute (PDA TRI) is offering seven courses in conjunction with this meeting – covering topics such as GMP regulations, Rapid Micro Methods, six sigma in process validation, cleanroom management and more!

EMA Perspective: Knowledge Management Fostering Academic-Industry Collaboration for Product Development

M. Lynn Crimson, Dean, School of Pharmacy, University of Texas

Janet Walkow, PhD, Director, Drug Dynamics Institute, University of Texas

Piotr Krauze, Scientific Administrator/Compliance and Inspection Sector, European Medicines Agency

Ghulam Shabir Arain, PhD, FCQIPrincipal ScientistAbbott Laboratories, UK

Douglas J. Ball, M.S.Diplomate, American Board of Toxicology; Research Fellow, Drug Safety Research & DevelopmentPfizer Global Research & Development

Suraj Baloda, Ph.D.Director, QC Microbiology & Environmental ControlBen Venue Laboratories, Inc.

Rory BudihandojoDirector, Quality Systems AuditBoehringer Ingelheim Shanghai Pharmaceuticals Co., Ltd.

Suggy S. Chrai, Ph.D., MBA., R.Ph.President & CEOChrai Associates, Inc.

Melanie W. CookeAssociate ScientistAmgen, Inc.

Emil W. CiurczakChief Technical OfficerCadrai Technology Group

Rick E. CooleyNorth America Manager of Process AnalyticsDionex Corporation

Walter Dziki, Ph.D.Associate Research FellowAbbott Laboratories

John Finkbohner, Ph.D.Director, Regulatory AffairsMedImmune

Adam S. GoldsteinSenior Manager, Clinical Purification Operations/Development Genentech

Brian Lingfeng HeResearch InvestigatorBristol-Myers Squibb

Ronald G. Iacocca, Ph.D.Research AdvisorEli Lilly and Company

Thomas A. Jennings, Ph.D.Founding PresidentPhase Technologies, Inc.

Maik W. JornitzGroup Vice President Marketing Fi/FRISartorius-Stedim, Inc.

Hemant N. Joshi, Ph.D., MBAPrincipalTara Innovations LLC

Daniel R. KlevishaDirector of Pharmaceutical & Chemical SalesThermo Fisher Scientific

Ian Lewis, Ph.D.Marketing ManagerKaiser Optical Systems, Inc.

Jack LysfjordPrincipal ConsultantLysfjord Consulting LLC

Steven R. Maple, Ph.D.Head of Pharmaceutical Technology Development Dept.Lilly Research Laboratories, Eli Lilly and Company

Jerold M. MartinSenior Vice President, Scientific AffairsPall Life Sciences

John P. MayerSenior Research AdvisorEli Lilly and Company

Mark McDowall, Ph.D.Strategic Development Manager, Mass SpectrometryWaters Corporation

Theodore H. Meltzer, Ph.D.Capitola Consultancy

Ronald W. Miller, Ph.D., M.B.A.President , Technology ConsultantMiller Pharmaceutical

Ganapathy Mohan, Ph.D.Sr. Director, Global Analytical Sciences Dept.Sanofi Aventis

Shane R. Needham, Ph.D.Laboratory DirectorAlturas Analytics, Inc.

Daniel L. NorwoodDirector, Physical and Chemical AnalysisBoehringer Ingelheim Pharmaceuticals, Inc.

Yashwant Pathak, Ph.D.Asst. Dean Academic AffairsProfessor & Chair, Dept. of Pharm. SciencesCollege of PharmacySullivam University

Saeed A. QureshiSenior Research ScientistHealth Canada

David RadspinnerDirector of Marketing and Applications Support for BioProcess ProductionThermo Fisher Scientific

Aniruddha M. Railkar, Ph.D.Assistant DirectorSynerx Pharma LLC

Gary E. RitchieSenior AssociateLachman Consultants

Rodolfo J. Romañach, Ph.D.Professor of ChemistryUniversity of Puerto Rico, Mayagüez Campus

Jim RydzakInvestigator, Strategic Technology DivisionGlaxoSmithKline

Paul SheskeyDevelopment Leader in the Water Soluble PolymersDow Chemical Company

Charles A. Signorino, Ph.D.CEOEmerson Resources

Donald C. SingerGlobal Lead Manager, Microbiology, R&D GlaxoSmithKline

Onkar N. Singh, Ph.D., M.B.A. Assistant Director, Development Pharmaceutics Research & DevelopmentAlcon Research Ltd.

Arjen P. TinkePrincipal Scientist, Particle CharacterizationJohnson & Johnson Pharmaceutical R&D

» Editorial Advisory Board »


124 Bernard E. Saint Jean Drive, East Falmouth, MA 02536 USAtel: 888.395.2221 • fax: 508.540.8680 • [email protected] • www.acciusa.com

Run Both Chromogenic AND Turbidimetr ic Assays in

The Same K inetic Tube Reader

F L E X I B I L I T Y

CovEr FEAturEs

12 InformaticsThe Adaptable Laboratory: A Holistic Informatics ArchitectureJames M. Roberts, Ph.D., Mark F. Bean, Ph.D., Chris Bizon, Ph.D., John C. Hollerton & William K. Young, Ph.D., GlaxoSmithKline

22 Powder X-ray Diffraction (PXrD)Approaches to Quantification of Amorphous Content in Crystalline Drug Substance by Powder X-ray DiffractionPeter Varlashkin, Ph.D., GlaxoSmithKline

36 ramanCalibration of Multivariate Predictive Models: The Study of Factors Influencing the Prediction Accuracy of Raman Spectroscopy Applied to Pharmaceutical TabletsJoanny Salvas, Jean-Sébastien Simard, Ryan Gosselin, Ph.D. & Nicolas Abatzoglou, Ph.D., Pfizer

58 single useTransfer, Implementation and Late Stage Development of an End-To-End Single-Use Process for Monoclonal Antibody ManufactureBrian Mullan, Ph.D., Kristi Huntington, Aidan Collins, and Marie Murphy, Ph.D., Eli Lilly & Co.

66 Formulation DevelopmentAmorphous Solid Dispersions as Enabling Formulations for Discovery and Early DevelopmentBrian E. Padden, Ph.D., Jonathan M. Miller, Ph.D., Timothy Robbins, Ph.D., Philip D. Zocharski, Leena Prasad, Julie K. Spence & Justin LaFountaine, Abbott Laboratories

January/February 2011 | Volume 14, Issue 1

January/February 2011 | | 7

30 PArtIClE sIzIng

Physical Characterization of Nano Particulates Used in the Pharmaceutical IndustryRon Iacocca, Ph.D. Eli Lilly & Company

44 DIssolutIon

Limitations of Some Commonly Described Practices in Drug Dissolution Testing and Suggestions to Address TheseSaeed A. Qureshi Health Canada, Banting Research Centre

50 mICroBIology

Case Studies of Microbial Contamination in Biologic Product ManufacturingKalavati Suvarna, Ph.D, Anastasia Lolas, M.S., Patricia Hughes, Ph.D. & Richard L Friedman, M.S. Food and Drug Administration.

74 PrEFIllED syrIngEs/ EXtrACtABlEs & lEAChABlEs

Extractable and Leachable Implications on Biological Products in Prefilled SyringesFYasser Nashed-Samuel, Dengfeng Liu, Kiyoshi Fujimori, Lourdes Perez & Hans Lee, Ph.D. Amgen Inc.

82 EXCIPIEnts

Selling the Audit Observation to Enhance ConformanceIrwin B. Silverstein, Ph.D. IPEA

sPECIAl FEAturEs

86 Pittcon Advertiser Profiles

91 Aguettant Q&A

rEgulAr FEAturEs

90 Industry news

95 Classified Advertisements

96 Advertiser’s Index

» In thIs IssuE »


SINGLE-USE TECHNOLOGY

Single-use, reusable or hybrid.The right solution for each process step.

Sartorius Stedim BiotechUSA +1.800.368.7178 | Europe +49.551.308.0

You choose: single-use, reusable or a combination of both. You set the targets – we provide the technologies to reach them. Different product types, scale-up levels and development stages call for

different solutions. Together, we will simulate your processes and custom-engineer what best meets your needs. The result: maximum process reliability, high flexibility and optimized cost.

www.sartorius-stedim.com/single-useturning science into solutions

Welcome to the January/February issue of American Pharmaceutical Review!I hope 2011 is treating all of you well so far, and that everyone is surviving the winter storms out there!

I am not one for New Year’s resolutions, mainly because I give up or give in by February 1st, so in typical “me” fashion, this year was no different, no specific resolution for me, at least not at first. I vowed to have no promises to go to the gym every day or to go on a no-carb diet that will not be fulfilled because, quite frankly, I figure I should always make sure I exercise regularly and have healthy eating habits, right?

But this year, I did make a different kind of expectation for myself. I made a commitment to myself to make this year a better year than the ones in the past. Now that may seem vague, and honestly it is, but my goal is to simply be better, do better, and give more. Why this pertains to all of you is because this also means that I plan, along with my colleagues here at Russell Publishing, to make American Pharmaceutical Review an even better publication than it has been in the past. My hope and job is to always have the best editorial for our readers, and I will make sure that we continue to do that tenfold. As always, we are open to your comments and suggestions. Please let us know how we are doing!

I think we are starting 2011 off right with a great first round of editorial. James Roberts et al from GlaxoSmithKline are bringing you their next installment of their two-part series on informatics in the laboratory. In their previous article appearing in our 2010 September/October issue, they supported a laboratory solution that contained both simple and modular applications. Their focus was on the challenges that arose and what is still needed in analytical laboratories. With this in mind, their second article focuses on how laboratories can get to where they need to be. Read about their solution on page 12.

On page 58, Brian Mullan et al from Eli Lilly & Co. present an alternate route to biologics manufacturing that creates options for leveraging existing facilities, as opposed to constructing dedicated biologics manufacturing space. This is a must read for all Single-Use enthusiasts!

Brian Padden et al from Abbott will discuss their approach for screening and manufacturing amorphous solid dispersions for application in discovery and early development in their article, “Amorphous Solid Dispersions as Enabling Formulations for Discovery and Early Development.” Read all about their work on page 66.

Next up from the University of Sherbrooke is Nicolas Abatzoglou et al on page 36. The engineering goal of their work was to investigate the development protocol of MVPM-based PAT methods in order to identify the factors that are most influential on the performances of the developed method, and thus, to help proposing an optimized development protocol.

And finally, Peter Varlashkin from GlaxoSmithKline has contributed an article that will briefly mention the various PXRD approaches to quantify amorphous content, but for the most part, his focus is on a simple approach that does not require standards and is suitable for routine pharmaceutical development. Check it out on page 22.

Please take a look at the Pittcon Conference and Expo advertiser preview on page 86. I am very much looking forward to seeing many of you in Atlanta next month!

Kind Regards,

Emily M. [email protected]

"My hope and job is

to always have the best editorial for our readers,

and I will make sure that we continue to

do that tenfold"

» A message from the Editor »Emily M. JohnsonEditor, American Pharmaceutical Review


Where do the key players in pharma, Biotech and manufacturing come together?

March 29 - 31, 2011 | jacob k. javits center | new york, new york

INTERPHEXConnects | www.INTERPHEX.com

Interested in exhibiting at INTERPHEX 2011? Contact Pete Zezima at 203.840.5447 or [email protected]

»


InFormAtICs »

James M. Roberts1, Ph.D., Mark F. Bean2, Ph.D., Chris Bizon3, Ph.D., John C. Hollerton4, William K. Young5, Ph.D.

GlaxoSmithKline, Product Development1 Research Triangle Park, NC, USA5 Stevenage, Hertfordshire, UK

GlaxoSmithKline, Molecular Discovery Research2 Collegeville, PA, USA4 Stevenage, Hertfordshire, UK

Renaissance Computing Institute, University of North Carolina at Chapel Hill3 Chapel Hill, NC, USA

The Adaptable Laboratory: A Holistic Informatics Architecture

In a previous paper, we advocated a laboratory informatics solution comprised of simple, modular applications rather than currently available “big box” solutions; several examples of efficient and novel capabilities were provided [1]. Two themes arose. First, limited-scope, modular applications like those commonly used with personal computing devices make software easier to use. Second, facile extensibility of existing software is required to address unanticipated needs. The content of [1] focused largely on defining what the challenges of usability and extensibility actually are and what analytical laboratories need. This paper focuses on how laboratories might get there, how laboratory software can follow personal computing capabilities by months, not years or decades. The picture presented is a holistic one, not relegated to the domain of only a chromatography data system or only an electronic notebook or only a laboratory information management system: it is a foundation that can be used by all of these products as well as for new unforeseen application solutions. The specific architecture described is not dogmatic; it is but one solution, not necessarily the only solution. The principle, however, is stated axiomatically: analytical laboratories must adapt to unanticipated needs using data-driven software solutions.

Despite advances in automation and instrumentation, it is inevitable that customers encounter problems that the vendors have not provided for. Examples include validation of text input against local business rules (e.g., project codes, lab notebook references) and vendor-neutral solutions. Such problems, although related to the analytical instrumentation, generally lie outside the purview of vendors, yet they are problems that customers need to resolve. This paper attempts to suggest an architecture that permits unobstructed, flexible data access, allowing customers to resolve some of their own problems. It is an immediate continuation of [1]; the two papers should be read together and the figures compared side-by-side. While the discussion has focused on chromatography and the pharmaceutical industry, the principles and architecture should be well suited to other market sectors.

Adaptability in Biology and laboratory InformaticsThe laboratory informatics solution described here has similarities to information flow in biological systems. To illustrate computer concepts for a broader audience, the architecture is discussed in comparison with various components in biological systems. While the analogy does not bolster the proposed architecture, it is hopefully instructive. Like most analogies, it holds well at a high level but it should not be taken too far.

Biological systems are complex and information rich, far more so than our laboratory information systems. And yet biology works most of the time. Organisms are highly optimized for their environment and emergent properties arise ‘magically’ from information stored in DNA [2]. Laboratory

PA83

8212

10_L

New Core-Shell Phases

REVEALED!

© 2

011

Phe

nom

enex

, Inc

. All

right

s re

serv

ed. K

inet

ex is

a r

egis

tere

d t

rad

emar

k of

Phe

nom

enex

, Inc

.

See Them in Action! www.phenomenex.com/revealPhenomenex products are available worldwide. Email us at [email protected].

Ultra-high performance on ANY LC system just got more powerful. Introducing two exciting new phases to expand the range of core-shell selectivity! Kinetex® columns are now available in five selectivities: C18, HILIC, PFP, XB-C18, and C8.

XB-C18 C8

PA83821210_L_3.indd 1 1/28/11 10:48:25 AM

»


InFormAtICs »

informatics systems exhibit similar outcomes; powerful software and capabilities arise from files on disk or content stored in memory. Table 1 provides a list of hierarchical components that “build on each other” to comprise biological and informatics systems. The similarities lead to one conclusion: that of an adaptable organism or laboratory.

A holistic Approach to laboratory InformaticsThe overarching theme is that adaptability requires unprecedented access to the DNA of the laboratory. Throughout this paper, XML is cited as the document format of choice (the DNA of the lab). This is not prescriptive; other data-interchange formats like JSON (www.json.org) or open binary formats might be preferable in some situations. For brevity, however, XML is cited merely to illustrate the features and benefits of an open-document based solution. Figure 1 illustrates a holistic and adaptable computer architecture designed to meet all of the efficiencies and new capabilities described in [1] (e.g., point-and-click reports, modeling, optimizations) as well as new, unforeseen applications. At the highest conceptual level, the flow of information (data and instructions) starts from the bottom left corner of Figure 1. Upon opening any of the modular client applications, data is validated before entry into the system and then flows seamlessly through the

entire system in a clockwise direction back to the client application. In all cases the flow of information is mediated by specialized services and relational databases.

In summary, data is protected and traceable, satisfying 21 CFR 11 requirements [3]; data is validated before entry into the system to support unprecedented data mining and downstream error prevention capabilities and the architecture is rapidly extensible to support development of new ad hoc solutions or production-quality applications. Closed, vendor-specific data formats are not precluded, but they are also not required because all data is stored in XML. Finally, data is easily accessible from client applications and, importantly, from outside these applications using either SQL or XPath queries. Examples follow, but first a more detailed description of the architecture.

A Universe of Possibilities: The Primary Data StoreThe bottom right region of Figure 1, termed the Primary Data Store (PDS), contains all the laboratory data and instructions, literally everything that can be known about the system. The XML documents in the PDS are immutable: once created they can never be changed. They document the state of every part of the system at a specific point in time. New XML documents can be created to supersede existing ones, but existing documents cannot be modified or replaced.

The XML documents are produced by the modular application groups discussed in Figure 3 of [1] and represent sample metadata, analysis

Table 1. Similarities between biological and laboratory informatics systems in the architecture described here. The analogy is

hopefully instructive for those less familiar with computer systems. The key point is that both systems can lead to an adaptable

entity that can change to meet the needs of an ever-changing environment.

Biological Component

Informatics Component

Description

DNA XML Contains all the data and instructions, literally everything that can be known about the state of the system. Immutable: portions are copied and used by other components in the system, but never modified.

Gene XML Node A piece of data or instructions. Not all genes/nodes are used, but they are all accessible for potential future use by RNA/services.

DNA Repair Data Validation Corrects mistakes at the source.

Regulatory Genes Business Rules Controls information flow through the system.

RNA Services Translates data or instructions into a useful form.

Proteins Relational Databases Specialized components that efficiently carry out a very specific, useful function at high speed. Single-purpose, but interacts with others of its kind.

Cellular Wall Security Protects components from unauthorized access.

Differentiated Cell Modular Application Orchestrates activities of a variety of components to create a specialized, useful single-purpose function.

Adaptable Organism Adaptable Laboratory The collection of all components from which emergent properties arise. Each component can be rapidly adapted to meet the needs of a changing environment.

Figure 1. An adaptable architecture supports unanticipated and changing needs. Every time a modular application is opened

(purple box), data is checked against a Business Rules & Templates Repository (BRTR). Valid data then flows through modular client

applications into the Primary Data Store (PDS). Specialized services (orange) respond when new data arrives in the BRTR or PDS, extracting and placing only the necessary parts into

specialized relational databases that provide organized content to various client applications. The BRTR and PDS employ a shared,

public ontology; all other components are vendor-specific. SQL and XPath interfaces support ad hoc queries. Chromatography

Data Systems, Electronic Notebooks, and Laboratory Information Systems are all subsets within this architecture.

The new Chromeleon® 7.1 Chromatography Data

System brings Operational Simplicity™ to the Enterprise. Fully network all your LC/GC/IC instruments and realize the benefits of multiple intelligent features. Innovative eWorkflows enable anyone to run SOPs perfectly with just a few clicks, and the SmartPeaks™ integration assistant delivers the baselines you want in seconds.

From Samples to ResultsQuickly and Easily

Learn more at www.dionex.com/chromeleon7

Chromeleon 7 Simply Intelligent

Chromeleon is a registered trademark, and Operational Simplicity and SmartPeaks are trademarks of Dionex Corporation. PIN 1042

Pittcon 2011

Register for Dionex Courses

Carbohydrate Analysis or Capillary IC

www.dionex.com

/pittcon11

»


InFormAtICs »

metadata, analytical methods, laboratory instrument instructions, raw instrument data, processed data, and reported data. Binary data (e.g., the raw signal) may be encoded in Base64 and stored as a text string in the XML document or for larger data sets the XML document might contain a reference to binary data stored in an open, accessible format to support high-speed access.

Each XML document is uniquely named with a system-generated human-readable file name. For example, the XML document representing processed data might be named as a concatenation of the unique instrument ID, a timestamp identifying the start of the data acquisition, and a timestamp representing the start of data processing. This represents an interpretable and unique file that can be quickly found by the services and its data extracted and used by other parts of the system. Other naming schemes are possible, but they must ensure a globally unique file name. As with DNA, not all XML nodes (genes) are useful for all parts of the system. However, each node is easily accessible to all other parts of the system. This facile accessibility is currently missing in today’s laboratory and it is the fundamental key to adaptability discussed in the examples below.

Invisible Helpers: Specialized ServicesA service is an invisible software application that runs “in the background.” There are two kinds of services (orange box): the minority that interact directly with a client application (dashed line) and the majority (solid line) that monitor the creation of new XML documents on a secure file share and respond accordingly by safely extracting useful portions and placing them in a limited-scope relational database (blue box). This data translation requires a well-defined object-relational model that is more easily developed if the relational databases are kept manageably small and of limited scope. Keep it simple. The services themselves will vary in complexity depending on scope of the solution, but all of them will likely implement event-based monitoring and multiple threads to ensure adequate performance (e.g., filling the database nearly instantaneously upon arrival of the XML document into the PDS). Note that in addition to simply shuffling data through the system, these individual services can be licensed as distinct products and combined in numerous ways to form configurable workflows using a workflow management application.

Fast Data Access: Specialized Relational DatabasesFueled by the services that provide data from the PDS and BRTR, the relational database management system (RDBMS) provides fast access to all the data used by all applications. Two exceptions to this rule are high-performance applications where a service might provide content directly to the client from an in-memory cache, bypassing the RDBMS, or when “live” streaming data is required (dashed arrow in Figure 1). Each database stores all superseded records, but by default applications might expose only the currently valid data from the PDS and BRTR to prevent cluttering the work environment with invalid content (e.g., superseded instrument methods). Thus, audit trails and traceability are available upon special request but “old data” does not clutter the current working environment. In principle, a new database can be easily added, built from the primary data stored in the immutable XML documents stored in the PDS.

Keeping Data Safe: Security, Authentication, AuthorizationPrimary data in the XML documents are protected by the computer’s operating system. Information stored in the XML documents is accessible, but only by services and a select few individuals with appropriate authorization. Modern operating systems provide this level of security by authenticating (you are who you say you are) and authorizing (yes, you may have access) user access to content stored in the XML. Additionally, modern scalable frameworks like iRODS (Integrated Rule-Oriented Data System), used by the digital archiving, seismology, astronomy, and physics communities, offer powerful security for file-based collections using granular, policy-based rules that determine who can do what with a given file [4].

Poka yoke – getting it right the First timeToday’s vendor applications rarely provide sufficient validation of data entry. Manual, unchecked keyboard entry of text is ubiquitous and this means that databases are filled with incorrect data or 50 different correct ways to say the same thing; both scenarios result in nearly useless database queries. And this makes getting meaningful data from the RDBMS nearly impossible. Data validation is implemented here with a “poka yoke” (mistake proofing) mindset, a term borrowed from lean manufacturing in which errors are ideally corrected before they have a deleterious effect. The bottom left corner of Figure 1 depicts this feature implemented using the Business Rules & Templates Repository (BRTR).

Like data in the PDS, these individual XML documents are also immutable, but updateable with new unique and time-stamped versions to support traceability. Unlike the PDS, however, the XML documents in this validation section are singletons: one and only one document instance is valid at any given time and it is the “master” that client applications must consume and conform to every time the application is opened (although content could be mirrored across multiple computers). It effectively serves the role of a “benevolent dictator”, protecting the system from inadvertent mistakes, because only valid objects are presented to the user’s client for downstream entry into the PDS and relational databases. Structurally, this could be implemented as a dictionary containing key-value pairs. The values can be simple or complex depending on how many variables are needed to describe the object. For example, the values for an HPLC column might contain one and only one validated value for each of the following: vendor, model, part number (not serial number), length, diameter, particle size, stationary phase, and minimum and maximum values for pH and temperature. Validated values for these objects leads to extremely positive downstream affects: fast meaningful aggregation queries, intelligent error checking in the lab, instantaneous global adoption of business rules, and point-and-click report generation. Four control mechanisms exist in the BRTR and are summarized in Table 2.

Job #: WATR13531_A

Job Name: WATR13531_A_Bioanalysis_DBmec_edit.indd Date: 01-26-11Live: 6.75"x9.5" SmallTrim: 7.75"x10.5” LargeTrim: 8.375"x10.875" Bleed: 8.625"x11.375" Page: 1 Rev: 1Stage: mech Release: 01-26-11 DB AD: CP CW: GK PM: JT Scale: 100%Color: 4C Gutter: — Other: — ST: DB/JC

MK: _____________ PM: ______________ AD: ______________

CW: _____________ QA: ______________ CD: ______________

Initials / Date Initials / Date Initials / Date

Client / Date: ________________________________

MECH

PROOF

APP

RO

VALS

PUBS: American LaboratoryAmerican Drug DiscoveryAmerican Pharmaceutical ReviewBio BusinessBio IT WorldBioTech WorldBioTechnology FocusBioPharm InternationalBioProcess InternationalBioSpectrum AsiaChemical & Engineering NewsDrug Discovery and Dev

Drug Discovery NewsEnvironmental Science & TechnologyFood ProcessingFood QualityFood Safety MagazineGenetic Engineering News (GEN)Genome TechnologyGenomics & ProteomicsJour of The American Soc for Mass SpectrometryLab Asia Media GuideLab Business - JesmarLCGC AsiaLCGC N. America

LCGC EuropeMolecular and Cellular Protemomics Nature MethodsPharmaceutical Discovery & DevelopmentPharmaceutical ManufacturingPharmaceutical TechnologyPharmaceutical Technology EUROPEPharmaceutical ExecutivePharm Form and QualityProteomics JournalScientific Computing & InstrumentationScientific Computing World

Please note this is a COMMON SIZE mechanical file, you will need to center file using the center marks

provided when placing ad in the publication page area. (common size = smallest live/smallest trim / largest bleed)

©2011 Waters Corporation. Waters is a registered trademark of Waters Corporation. The Science of What’s Possible is a trademark of Waters Corporation.

Have you seen the world’s most comprehensive Bioanalysis system solution? Waters has introduced the technology to tackle any challenge, as well as

transform your laboratory. And now we’re taking it on tour.

»


InFormAtICs »

Adaptability in Action: Examples from the real World

Instrument Control & Data AcquisitionReal-time, closed-loop automated control of laboratory instruments was described in [1]. These and other “intelligent” applications require a new level of instrument control. Figure 2 shows how this architecture can support rapid development of new experiments in the lab, where the instrument is treated as a “black box” that receives instructions from Application Group 2 [1] in the form of an XML document containing a sequence of instructions.

In this model, the primary job of the laboratory instrument is to execute instructions provided by the XML document, raise events to notify clients that something important has happened, and raise exceptions if something goes wrong so clients may handle errors appropriately. For example, if the instrument is an NMR, the instruction set might be sample metadata along with a series of shaped RF pulses, delays, and acquisitions combined to make a novel pulse sequence; for HPLC, one single instruction could be “turn on column heater and heat to 40 degrees C.” All of these instructions are mediated by the instrument-vendor’s instrument control service that reads the simple XML instructions and implements a max-priority queue [5] of sample

sequences, bundled with the instrument instructions associated with each sample in the sequence. All the complexity of instrument control is hidden and handled by the service. The idiom is, “Here’s the sample and it’s method information, go!” This approach amounts to a facade design pattern [6, 7] that simplifies today’s proprietary instrument drivers and is akin to a specification-implementation model used for computer language compilers. Different analytical techniques will obviously require different instruction sets and vendor-specific hardware will require slight differences in instruction sets based on different hardware features or capabilities. However, all instructions should be defined at a level of abstraction to support adaptable work practices by analytical chemists, not software developers.

Data acquisition is handled by the instrument vendor’s service and has two distinct outputs. First, during data acquisition a typed (decorated) binary stream is accessible to enable reading of temporary, non-stored “live” data as it is generated from the instrument. Second, upon completion of data acquisition, the raw data produced by the instrument is written to the PDS in “Schema Group 3” (see Figure 1).

This level of instrument control and data acquisition allows for intelligent, live, interactive method development, optimization, and error correction on-the-fly, without the need for manual intervention. This automated closed loop of “control instrument – analyze data – control instrument” offers unprecedented opportunity to automate extremely time-consuming manual activities (e.g., optimization) and it is easily supported using this approach.

Data Access – ConceptTwo sources exist for ad hoc data access: the relational databases and the XML documents. This two-option data access model exploits the strengths and avoids the weaknesses of both XML and RDBMS as data storage mechanisms. For example, XML encapsulates

Table 2. Business rules are enforced globally and immediately before data enters the system because each client application

checks for updates to configurations, business rules, and templates every time the application is opened for use. This “poka yoke”

error prevention and correction enables instant adoption of changes in business rules and SOPs across an entire organization.

Control Mechanism Purpose Examples

Application Configuration & Installation Packages

Ensure that the correct version of software is used and role-based permissions are enforced.

application version, user roles and authorization, granular permissions, installation packages to update applications

Content Validation Ensure that business rules are applied uniformly and immediately.

analytical techniques, analytical methods, instrument identifiers, sites, buildings, laboratories, addresses, people, teams, date and time formats, projects, columns, packaging materials, storage conditions, reagents, solvents, batches, chemical structures, calculated chemical properties, physical constants

Analysis Template Ensure that all experiments are carried out according to appropriate scientific and business standards.

purpose-conclusion, method development, method validation, dissolution, content uniformity, excipient compatibility, solubility, stability, reaction monitoring, identity, purity, assay, structural elucidation, method transfers, robustness, ruggedness, disintegration, friability, particle size distribution, heavy metals, loss on drying, residual solvents, ash, water content

Report Templates Ensure that dynamically generated point-and-click reports conform to currently accepted internal business rules or external guidance.

certificates of analysis, batch impurities, stability, column logging, instrument utilization metrics, IND, NDA, IMPD, MAA

Figure 2. Specification-implementation model for laboratory instrument control and data acquisition. Instrument vendors

implement instructions provided to the instrument control service by Application Group 2. Data is saved to the PDS as raw,

unprocessed data for consumption by Application Group 4 (Data Processing, see [1] and Figure 1 above). Live data streaming

from the instrument is provided as a decorated stream for client applications to display and respond to live data.

A Clear Road Ahead for Metabolite IDComplimentary technology for your evolving drug metabolism needs.

Whether your needs revolve around low-level metabolite identification or high resolution accurate mass structural elucidation,

the AB SCIEX QTRAP® 5500 and the TripleTOF™ 5600 mass spectrometry systems can make your drug metabolism goals a reality.

See how clear the road ahead for metabolite ID can be with targeted predictive MRM and non-targeted full scan MIMs approaches.

Discover how HRMS Qual-Quant workflows can accelerate and simplify complete metabolite characterization and metabolic

clearance in a single analysis.

For more information, visit www.absciex.com/applications/drugmet

For Research Use Only. Not for use in diagnostic procedures.© 2010 AB SCIEX. The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners. AB SCIEX™ is being used under license.

AB SCIEX TripleTOF™ 5600 SystemAB SCIEX QTRAP® 5500 System

»


InFormAtICs »

everything there is to know about a single object – one sample, one sample sequence, one experiment, one instrument, etc. – in a single document. This gives rise to its primary strength – all the data that describes the object is accessible from one neat package and XML can be intuitively structured to access anything and everything from the single XML document. XML is also descriptive; it conveys meaning and is easily interpreted. So if you want to know everything about a few objects, simply use one of many available third-party tools to query the XML. However, XML is not ideal for working with extremely large collections of similar objects. Enter RDBMS, which organizes data in tables and provides superior data access to large collections of similar objects, spread across many different tables. The primary benefit of this organization is the ability to execute arbitrary and unforeseen queries across multiple objects in an efficient manner. The weakness of RDBMS, however, is that without clear documentation describing the database, it can be difficult or impossible for a non-specialist to interpret, and therefore very difficult to actually create new queries. While the greater generality and efficiency associated with multi-object queries often tips the tables towards exclusive use of RDBMS, domains in which data needs are well understood, and which are primarily single-object based (as with many laboratory activities) can benefit from the usability of the document format. Furthermore, a document model offers a convenient way to aggregate data, e.g., from multiple vendors. If vendors each create a separate database, it can be technically challenging to query across these databases; a document-based aggregation, though it may be somewhat inefficient, is trivially achieved. In the examples that follow, this pattern of moving between RDBMS and XML data formats emerges as an extremely flexible solution to the adaptability challenge facing today’s laboratories.

Data Access – ExampleIt’s a common scenario. A manager or scientist asks a question that requires data spread across multiple systems or locations. Figure 3 illustrates a rapid workflow that can provide fast answers to unanticipated questions, based on using the SQL and XPath interfaces specified in Figure 1.

If the necessary data exists in the RDBMS, then an ad hoc query is executed and an ad hoc result set is produced to satisfy a one-off need, usually in a few minutes. Custom queries will proliferate and over time these read-only SQL queries will form a collection that can be monitored for frequency of use or relative importance. If necessary, a stored procedure is easily added to the RDBMS, thus enabling a generalized answer that can be executed in seconds.

The second possibility is that the required data is not stored in the database (the “No” path in Figure 3). In this case, the PDS provides the data. Individual XML documents on the file share can be iterated over by operating system services and needed information contained in each XML document can be extracted using an XPath query. This pattern supports both DOM and SAX models, optimized for either full-control or high-speed data access. The end-result is the same as if the data were stored in the RDBMS: an ad hoc result set is provided quickly. As with the RDBMS data access example, frequently requested XPath queries can be prioritized and implemented as a new data service, relational data

table(s), or even a new database, each of which would be optimized to support the custom (but important) data access needs. To continue the biological analogy, introns (previously unused XML nodes) have become exons (useful XML nodes).

Point-and-Click ReportsWriting reports and documents manually is extremely time-consuming. This architecture supports the rapid development of new reports that can be generated with a few mouse clicks. For example, an analyst responsible for creating a 6-month stability report for a batch of tablets might simply start to enter the tablet batch number in the reporting application in Application Group 5, which auto-completes the text entry midstream (because the values are validated). The analyst then chooses the 6-month time point from a drop down list and clicks a button to request the report, which is generated and provided in a few seconds (the data being aggregated on-the-fly by the database). Importantly, all the content of the report is valid, enforced by the current Report Template. Likewise, the presentation (table formats, significant figures, charts, fonts, headers, footers, etc.) also conforms to currently accepted criteria, enforced by the Report Template. In short, an arbitrarily complex report can be accurately created in a few seconds. Thus, the analyst stops spending countless hours finding data, transcribing data, formatting data, building charts, and creating reports manually. Likewise, managers stop tedious review of data transcription. Instead, time is spent understanding the meaning of the data, identifying trends or outliers, and taking action based on the impact the data has on the project.

Figure 3. Extensible data access using both relational databases and XML documents in the PDS. Unanticipated questions can be

quickly answered using a SQL query against a relational database. Subsequent ad hoc answers then accumulate over time; those that rise in importance can be implemented as a stored procedure for

fast data access. If the data does not exist in the database, the PDS is queried after which important queries then get supported by a

new database.

»


« InFormAtICs

What We need: getting from here to ThereIn this two-paper series, we have tried to be explicitly clear about new, untapped opportunities that await the laboratory. We have also described a potential mechanism (not the only one) to make these concepts a reality. The architecture proposed here is different from many of today’s laboratory software offerings. It is a more open approach to accessing data and controlling laboratory instruments, but it is also more modular with opportunities for new commercially licensed products. In this closing section, the main challenges and next steps toward adoption are summarized.

Open, Shared SchemaThe architecture proposed here requires a shared language, a foundation on which everything else rests: the content stored in the document-based PDS and BRTR. This amounts to a holistic, open and shared schema – a common definition used by all instrument vendors, similar in spirit to the open, vendor-neutral AnIML instrument data format [8]. Successful implementation and adoption of these standards will require iterative development and an unprecedented level of collaboration between leaders in industry, academia, instrument vendors, and regulatory agencies.

Closed, Proprietary ImplementationEverything above the BRTR and PDS in Figure 1 can, and should be, a vendor-specific, commercially licensed product. Competition at the level of services, databases, and modular applications will produce the best products for customers. Using the shared schema, software vendors are free to differentiate themselves and compete for customers at this new level: that of the modular application and service. From a vendor’s perspective, the smaller, modular applications can be configured and combined in nearly limitless ways to gain customers in new markets: CROs, small biotech and large pharmaceutical companies, academic and government labs, and different chemical industries can all have variations, using the same foundation.

Next Steps: An Open Discussion of Risks, Costs and BenefitsFrom an instrument software vendor’s perspective, there are several risks and benefits associated with adopting this approach: loss of closed data formats, loss of vendor-specific instrument control, “it’s been tried before.” We do believe, however, that there are valid financial incentives for moving in this direction, not the least of which is that someone will do it (consider the MP3 file standard and the music industry). These risk-benefit-cost issues are too complex to address in a short communication and the authors believe that these topics are best discussed in person. What is needed is a mechanism to bring together representatives from industry, academia, instrument vendors, and regulatory agencies to discuss how laboratories can move toward more open, scalable and adaptable work practices. Please contact us if you have questions or criticisms.

references1. Roberts, J.M., Bean, M.F., Cole, S.R., Young, W.K., Weston, H.E., American Pharmaceutical

Review, September/October 2010, p. 60 – 67.

2. Complexity: A Guided Tour, 2009, Oxford University Press, Oxford, Melanie Mitchell.

3. Code of Federal Regulations Title 21, Part 11.

4. Proceedings, iRODS User Group Meeting 2010: Policy-Based Data Management, Sharing and Preservation, Edited by Reagan W. Moore, Arcot Rajasekar, Richard Marciano.

5. Introduction to Algorithms, 2nd Edition, 2001, The MIT Press, Cambridge, MA, p 138 – 140, Thomas H. Cormen, Charles E. Leiserson, Ronald L. Rivest, Clifford Stein.

6. Head First Design Patterns, 2004, O’Reilly Media, Inc., Sebastapol, CA, p 254 – 270, Eric Freeman, Elisabeth Freeman.

7. Design Patterns: Elements of Reusable Object-Oriented Software, 1995, Addison-Wesley, Reading, MA, p 185 – 193, Erich Gamma, Richard Helm, Ralph Johnson, John Vlissides.

8. See http://www.scientificcomputing.com/the-iupac-astm-unified-standard.aspx.

Author BiographiesDr. James M. Roberts works in an automation group in Product Development at GlaxoSmithKline. He applies informatics, data integration, modeling, and automation to increase efficiency in analytical chemistry. He received a Ph.D. under the advisement of Professor Janet Osteryoung and has 12 years of experience in the pharmaceutical industry.

Dr. Mark F. Bean, Investigator at GlaxoSmithKline, has worked with automated MS and software solutions for research chemistry LCMS for 20 years. He was the architect-project lead for CANDI, a vendor-neutral LCMS login, processing, and results viewing suite used by GSK in the Philadelphia area. He is a founding member of the ASTM committee charged with the AnIML analytical data standard.

Dr. William K. Young is an Investigator in Analytical Sciences at GlaxoSmithKline in Stevenage, UK. He develops and maintains the walk-up chromatography systems within Chemical Development. He received his Ph.D. from Imperial College, London with Professor W. John Albery and has 11 years experience in the pharmaceutical industry.

John C. Hollerton is a Director of Analytical Chemistry at GlaxoSmithKline. He leads a team who offer spectroscopic and small molecule X-ray crystallography support to R&D as well as informatics support to Analytical Chemistry. He has spent the last 30 years integrating informatics in the analytical environment. He was responsible for the design and implementation of GSK’s Global Analytical Data Repository (GADR).

Dr. Chris Bizon is a Senior Research Scientist in Informatics at the Renaissance Computing Institute. He currently leads a group designing cyberinfrastructure for high-throughput genomic sequencing. He received his Ph.D. in physics at the University of Texas at Austin with Professor Jack Swift and has worked in both the academic and industrial settings

»


PXrD »

Peter Varlashkin, Ph.D. GlaxoSmithKline, US R&D

Approaches to Quantification of Amorphous Content in Crystalline Drug Substance by Powder X-ray Diffraction

IntroductionTypically, the desired solid state for active pharmaceutical ingredients (drug substance) is crystalline. Amorphous content within the crystalline drug substance may have a deleterious impact on the drug product such as reduced chemical and physical stability.

Powder X-ray diffraction (PXRD) is one of several significant tools used to characterize the solid state character of drug substances. Randall et al [1] discussed the various uses of PXRD to pharmaceutical development as applied both to drug substance and drug product.

Powder X-ray diffraction is considered the ‘gold standard’ for identifying and quantifying crystalline phases in materials as diverse as minerals to active pharmaceutical ingredients. For pharmaceutical samples with amorphous content within the range of approximately 10 to 90% w/w, powder X-ray diffraction (PXRD) provides a means to determine the relative amorphous content. While less-routine XRD techniques may provide a lower limit of detection/quantitation for amorphous content, in the author’s experience, 10% amorphous content is a practical lower limit for detection using typical PXRD equipment.

The purpose of this article is to briefly mention the various PXRD approaches to quantify amorphous content. The majority of this article will focus on a simple approach that does not require standards and is suitable for routine pharmaceutical development.

ExperimentalThe data presented were obtained with a typical Bragg-Brentano powder diffractometer using a copper X-ray source and a nickel filter at the detector to provide diffraction data using λ= 1.54 Å .

DiscussionDuring the development of a drug substance synthesis, initial material may be completely amorphous or poorly crystalline. PXRD can be used to screen for the relative crystallinity of samples. At some point, it may be advantageous to determine quantitatively the level of amorphous content in the samples.

»


« PXrD

Quantitative phase analysis by PXRD is not trivial [2]. However, the complexities of analyzing a material such as a geologic sample are lessened for organic samples such as pure drug substance or drug product due to minimal sample X-ray absorption affects.

Figure 1 shows the PXRD pattern for a mixture of crystalline and amorphous drug substance X. In a mixture of amorphous and crystalline material, the PXRD will exhibit both sharp and broad features. The sharp peaks are due to the crystalline component and the broad features (sometimes referred to as “halo”) to the amorphous component. Deconvolution of the mixture PXRD into separate PXRDs with sharp-only diffraction peaks and broad-only diffraction peaks will allow for determining the percentage of amorphous content.

Prior to applying quantitative methods to PXRD data for amorphous determination, the analyst must be sure that the broad features are due to amorphous (or glassy) material rather than due to microcrystalline or nanocrystalline material. The diffraction peak shape is affected by a variety of parameters, including the size of crystallites making up drug substance particles. As the crystallite size is reduced, the diffraction peaks broaden. Once the size is sufficiently reduced, the crystalline diffraction peaks may have broadened to the extent to merge into each other forming a single broad diffraction peak (halo). If a single-crystal X-ray diffraction data is available, the powder diffraction pattern can be calculated with different peak shapes and widths to simulate the PXRD of micro/

Figure 1: PXRD scan of sample X obtained using a back-filled cavity mount and a variable divergent slit. Scan collected in reflection

mode. The sharp features (peaks) are due to the crystalline component. The broad hump in the baseline is due to the

amorphous component. Background scattering unrelated to the amorphous and crystalline components produces an offset of the

baseline from 0 counts.

X-ray diffraction solutions for the pharmaceutical industry

EMPYREANWidest range of pharmaceutical applications: • Structural investigation of NCE’s • Polymorph and salt screening• In situ crystallization and scale-up studies• Investigation of amorphous and nano-crystalline

compounds• Stability and compatibility studies • Detection of low amounts of polymorphic

impurities • Determination of small to large amorphous content• Microstructure analysis of tablets by CT

For more information:[email protected]

The Analytical X-ray Company

Widest range of pharmaceutical applications:

Microstructure analysis of counterfeit and original tablet

100

2500

10000

22500

Inte

nsi

ty [

cou

nts

]

2 Theta [°]

20 30

10%30%50%90%

XRD scans for samples with different amorphous lactose content

Visit us at Pittcon booth 2261

PN7722.indd 1 04-02-2011 07:57:14

»


PXrD »

nanocrystalline material. Such material would be crystalline but less ordered due to the reduced repetition of molecular packing in 3-dimensional space compared to material exhibiting PXRD with sharp diffraction peaks. Analysis for microcrystalline material is a separate topic not discussed here. In the absence of single-crystal XRD data, the analyst should discuss with the sample submitter the history of the sample to understand if assaying for true amorphous content is appropriate. The presence of a glass transition in the differential scanning calorimetry profile of the material would serve as proof of the presence of amorphous material.

At the early stage and even later stages of development, it would be advantageous to use a method for determining amorphous content that did not require the use of standards. This approach is discussed next.

Approach 1 – PXRD Quantitation of Amorphous without the Use of StandardsFor samples where little material (tens of mg) is available, silicon or quartz zero background plates may be used. Typically samples are dusted on the plates previously coated with a thin film of petroleum jelly or other viscous material. The level of the petroleum jelly for PXRD sample preparation is typically kept low to minimize the signal from the petroleum jelly which might be confused for amorphous drug substance. For the analysis of amorphous content, the use of petroleum jelly should be avoided. Thus, the sample should remain horizontal during the diffraction scan which is a feature of theta/theta diffractometers. Theta/two-theta diffractometers require the sample to tilt with the scan which may cause the powder to fall off the stage. Sample spinning to improve counting statistics makes the problem worse so the use of theta/theta systems for zero background plate preparations is recommended.

Where more material is available (200 mg or greater), the sample material may be filled into a sample cavity. For quantitation of mixtures of crystalline phases, back-filling [3] is recommended to avoid the effects of preferred orientation of particles when the sample is filled from the top of the cavity and smoothed flat with a glass slide. For the quantitation of amorphous/crystalline mixtures using the complete diffraction pattern, preferred orientation issues are less important. Thus front-filling of the sample is an option if back-filling is not available or problematic. A comparison of the reproducibility of various sample preparations on the raw and processed diffraction data is recommended.

Modern diffractometers allow for the data collection to be obtained with either fixed or variable divergent slits. The slit situated in front of the X-ray source, limit the divergence of the X-ray beam. PXRD results collected using a fixed divergent slit is also known as “constant-volume” data. Starting from low two-theta and scanning to higher two-theta, the area illuminated by the X-ray beam gets smaller and the depth of penetration increases. Provided the area illuminated with X-rays does not exceed the sample boundaries and the sample depth is sufficiently thick that the X-ray beam will not penetrate to the bottom of the sample at the highest scanned diffraction angle, the sample volume interacting with the X-ray beam remains constant.

PXRD results collected using a variable divergent slit is also known as “constant-area” data. The divergent slit is decreased as the scan goes to higher diffraction angles which results in a constant area of sample illumination but different sample penetration by the X-rays with varying diffraction angle. For a thin zero background plate preparation, the X-ray beam will completely penetrate the sample such that both a constant sample area and a constant sample volume are analyzed.

For cavity-amounted samples, the fixed divergent slit operation may be preferred so that a constant volume is analyzed throughout the diffraction scan. For zero background plate preparations, the variable divergent slit approach is recommended. In practice, data can be collected both ways during method development.

Where sufficient sample is available, a cavity-filled sample preparation is recommended to maximize the amorphous signal relative to the background.

In addition to the typical reflection geometry, modern research grade diffractometers allow for the option of collecting the diffraction data in a transmission mode. This alternate mode may be considered during method development.

For routine PXRD, the typical diffraction scan range is around approximately 2 to 45 degrees two-theta (with copper K-alpha radiation). Most PXRD vendor software allows the user to deconvolute the PXRD pattern to isolate the crystalline component. A blank scan should be run to determine the background artifacts from the diffractometer. Depending on the diffractometer optics, the lower diffraction limit for integration of the diffraction signal from the amorphous and crystalline components will be approximately 3 to 4 degrees two-theta (to avoid scatter from the direct X-ray beam at low angles). The upper scan limit should be approximately 70 degrees to allow proper estimate of the PXRD baseline.

Figure 2 shows the resulting deconvolution of the sample PXRD scan presented in Figure 1. Different vendor software allow the user to adjust the resolution to discriminate between the gradual baseline changes due to amorphous content and background scatter. By subtracting out the portion of the data above the baseline that separates the crystalline from amorphous components, the diffraction intensity for the amorphous component can be determined. The ratio of the diffraction intensity for the amorphous component divided by the total diffraction intensity multiplied by 100 provides the approximation of the percent weight/weight amorphous content. The starting point (low two-theta) for integration is just above the point where scattering from the direct beam is observed. The ending point is where the diffraction signal returns to the background level.

The approach may result in background signal due to X-ray scattering by air and the sample holder being incorporated into the diffraction area associated with the amorphous component. The deconvolution process can be refined by running blank scans. For samples prepared using zero background plates, the blank scan would simply use a clean zero background plate with no applied sample. For samples prepared using cavity mounted sample holders, an empty sample holder is not appropriate since scattering from inside the sample holder may be

Innovation with Integrity

Analytical Power and Speed for: • Small Molecule Identification • Trace Component Analysis • Intact Protein Analysis • Peptide Identification and Sequencing • Characterization of Post Translational Modifications

X-RAY POWER for PHARMA:• Phase/polymorph identification and quantification• Non-ambient studies including computer-controlled humidity• Structure solution and analysis from powder data using TOPAS• High-throughput solutions from the XRD market leader

maXis D8 Advance

LC-MS / XRD

www.bruker.com/d8advancewww.bruker.com/ms

C

M

Y

CM

MY

CY

CMY

K

APR Bruker Dual Ad Full Page print.pdf 2/4/2011 4:28:42 PM

»


PXrD »

observed. If available, a zero background plate inserted into the cavity can serve as a blank. The height of zero background plate should be flush with the top edge of the cavity. Applying a small piece of modeling clay affixed to the underneath side of the plate and then turning the sample holder over and pressing against a smooth and clean surface (such as as a flat bench top or a glass slide) can help properly align the plate with the cavity top. Inspection of the blank scans would allow for estimating the instrumental background and then subtracting this from sample scan prior to deconvolution of the sample diffraction pattern into the crystalline and amorphous components. The PXRD scan presented in Figure 2 had the instrumental background removed prior to deconvolution into amorphous and crystalline components. The lower the percent amorphous, the more attention to detail in background removal and integration parameters is required.

If possible, a sample containing a sizeable fraction of amorphous content (30 to 70%) should be analyzed by 13C solid-state nuclear magnetic resonance (SSNMR). SSNMR is a “nuclei-counting” technique and does not require external standards to quantitate amorphous content. The results from the SSNMR can be used to check the PXRD results and aid in optimizing the PXRD processing parameters, the choice of sample preparation, the choice of variable or fixed divergent slits, and the choice of reflection or transmission data collection. The SSNMR results are most trustworthy when there is a clear separation in the resonance amorphous and crystalline peaks. If there is overlap requiring deconvolution to separate the amorphous and crystalline components, then there is added uncertainty in the percent amorphous calculation. If the overlap in the SSNMR data is severe, then SSNMR may not be useful in providing an orthogonal check of the PXRD method.

Vibrational spectroscopy (IR and Raman) can also be used to check the PXRD method. However, the IR and Raman methods require external calibration curves that would involve preparing mixtures of pure and amorphous standards. Lack of homogeneous mixing, particle size/morphology differences between the calibration curves and actual samples, and differences in the nature of the amorphous standard and amorphous content in the sample will lead to errors in the estimate of the percent amorphous.

Preparation of known w/w percent mixtures of amorphous and crystalline material can also be used to check the PXRD method with the same caveats as presented for analysis using vibrational spectroscopy. For example, an amorphous standard prepared by ball-milling, spray drying, or lyophilization may not be fully equivalent to actual amorphous content produced in the sample by some other process (e.g. micronization using an air-jet mill or fast precipitation of the drug substance during crystallization).

Approach 2 – PXRD Quantitation of Amorphous with External StandardsAs mentioned before, mixtures of amorphous and crystalline materials may be prepared. If a pure amorphous standard is not possible, a partially amorphous standard (e.g. 60%) may be used but would require estimation of the percent amorphous content by PXRD using Approach 1 or an orthogonal technique such as SSNMR. The same approach can be used if a pure crystalline standard is unavailable.

Each mixture can be prepared and then analyzed by method in Approach 1. The calibration curve would plot the percent amorphous, as determined by Approach 1 on the y-axis versus the weight/weight percent from the known weights of amorphous and crystalline material used to prepare the calibration mixtures. The calibration curve should be approximately linear allowing for a least squares fit. From the calibration curve, the observed % amorphous based on the area responses of the crystalline and amorphous components can be converted into a weight/weight percentage.

A simplified variation on Approach 2 is to use the peak intensity of a single XRD peak for the crystalline component and the intensity at a two-theta position where signal from amorphous content is present but there is not an overlapping crystalline XRD peak. The single XRD peak should be at a low enough two-theta value where significant signal from the amorphous is absent or the baseline intensity near the XRD peak can be used to subtract the amorphous signal from the XRD (crystalline) peak intensity. Preferred orientation and size of the diffraction domains of the crystalline component need to controlled [3]. With XRD deconvolution tools available, use of the full diffraction pattern is preferred.

With all the caveats using mixtures of external standards, the approach without standards is recommended. Many analytical results (e.g. % area-under-the curve by HPLC, particle size distribution) produce results that are relative and not absolute but still allow for batch-to-batch comparison and data trending.

Figure 2: PXRD scan of sample X following deconvolution of the diffraction pattern into the crystalline (portion above green trace)

and amorphous (portion below green trace) components.

»


« PXrD

Approach 3 – Use of Rietveld Analysis for Quantitation of AmorphousIf the single crystal X-ray diffraction structure is available, Rietveld-based methods that use the whole diffraction pattern may be employed to estimate the percentage of amorphous content in drug substance [2]. The approach would require spiking the sample with an internal standard where the crystal structure is known such as silicon powder. The data should be collected using a fixed divergent slit. The analysis involves calculating the diffraction patterns for the crystalline drug substance and the internal standard and then varying the relative amounts of each component until good agreement between the observed and calculated diffraction patterns for the spiked sample are obtained. The use of the internal standard allows the Rietveld analysis to provide the weight fraction of the crystalline component. Subtraction of this value from 1 would yield the estimated weight fraction of the amorphous component. Approach 3 can also be an alternate approach to check the results from Approach 1 or 2. The use of Rietveld approach for quantitation of amorphous should not be undertaken by the novice XRD user and thus no further details are provided. For the more sophisticated user, Rietveld tools are typically part of the software of modern research grade diffraction systems.

For samples with amorphous content below 10%, the difficulty in obtaining reliable results by PXRD increases and alternate techniques to assess amorphous content, such as gravimetric vapor sorption or thermal analysis, are suggested. For samples with crystalline material below 10%, Approaches 1 or 2 may be sufficient. For higher precision, standard addition methods by spiking the sample with crystalline material may be required.

ConclusionsPXRD represents a convenient method to determine amorphous content over a broad range. The use of the approach without standards is convenient and often sufficiently accurate to help drive the optimization of the drug substance synthesis.

AcknowledgementThe author thanks Graham Whitesell of GSK for reviewing this manuscript.

Enhance ProductivityPDF-4/Organics 2011

A comprehensive materials database featuring ~436,000 organic and organometallic phases.

Designed for rapid materials identificationPolymorph Screening

Quality ControlDrug & Excipients Identification

Formulation AnalysisQuantitative Analysis

Polymorph IdentificationCrystallite Size

XRPD, a nondestructivetechnique, coupled with ourcomprehensive database isuseful for preformulation,drug development, andpatent applications.

International Centrefor

Diffraction Data

Databases from the Database Experts

[email protected]

Phone: 610.325.9814Toll-free: 866.378.9331 (U.S. & Canada)

»


PXrD »

references1. Randall, C., Rocco, W., Ricou, P., XRD in Pharmaceutical Analysis: A Versatile Tool for

Problem-Solving

2. Powder Diffraction, Theory and Practice, Dinnebier, Billinge, S. (eds.), Chapter 11 (Quantitative Phase Analysis, authors Madsen, I, Scarlett, N.), The Royal Society of Chemistry, RSC Publishing (2008)

3. A Practical Guide for the Preparation of Specimens for X-ray Fluorescence and X-ray Diffraction Analysis, Buhrke, V., Jenkins, R., Smith D. (eds), Wiley, VCH, 1998.

BiographyPeter Varlashkin received his PhD in Analytical Chemistry from the University of Tennessee (Knoxville). He has been employed by GlaxoSmithKline (GSK) for over 22 years. He currently works in the Physical Properties group within GSK (RTP, NC, USA). He has published several papers on powder X-ray diffraction and is on the organizing committee of the Pharmaceutical Powder X-ray Diffraction Symposia as well as a member of the International Centre for Diffraction Data.

Become an author.American Pharmaceutical Review strives to inform readers of the latest information in the

pharmaceutical industry. If you are employed at a major pharmaceutical company or university, or are an established consultant in the pharmaceutical industry, you are exactly who we are looking for.

All articles submitted must be unbiased, not mentioning specific vendor companies or product names, and of a technical nature. For more information on becoming one of

our authors, contact us at 317-816-8787 or [email protected].

www.americanpharmaceuticalreview.com

Untitled-1 1 1/21/11 6:33:24 PM

»


PArtIClE sIzIng »

Ron Iacocca, Ph.D.Eli Lilly & Company

Physical Characterization of Nano Particulates Used in the Pharmaceutical Industry

AbstractNano particulates or nano-scaled pharmaceutical products have received great interest for the past two decades. Reduction in size to the nano scale can offer unique therapeutic advantages, such as the ability of particles to pass through cell membranes, thereby targeting specific cells. There has also been a great deal of research performed on the use of nano particles for imaging specific tissues or tumors in vivo. The physical characterization of these nano materials poses certain challenges to the scientist/engineer working in this field. This article highlights these analytical challenges and proposes a suite of analytical techniques, that, when used in conjunction with each other, provides a thorough characterization of the material.

IntroductionThe formal definition of nano particle is a discrete entity that has a dimension of 100 nm or less. A nano-scaled material, on the other hand, may be comprised of nano-scaled structures, though the physical dimensions of the material may exceed 100 nm. A spray-dried agglomerate of nano-sized particles would be described as nano-scaled. For pharmaceutical applications, one does not resort to the use of nano materials unless there is a therapeutic driving force to do so. If the compound is poorly soluble in-vivo or if the molecule is highly potent, and the desired presentation of the drug product is a tablet, capsule, or suspension, the decision may be implemented to produce the active pharmaceutical ingredient as a nano material.

The actual form of the nano materials covers a number of technologies, from actual nanocrystals of API, to emulsions, to liposomes. An excellent review of the current technology can be found in Ref. [1]. Regardless of the formulation or carrier, once this size range is implemented, the drug product performance is intimately connected to the physical properties of the active pharmaceutical ingredient. Physical characterization becomes a crucial part of the drug substance and drug product control strategy.

»


« PArtIClE sIzIng

Physical Characterization techniquesSample PreparationTo obtain accurate and meaningful particle size data, the challenge for nano materials lies in the sample preparation. Because the material is so small and so surface active, generally accepted scientific practices can often end up producing artifacts, or altering the sample during measurement.

Toxicity concerns for highly potent nanocrystals suggest that the material should not exist as a dry powder, but can be more safely handled dispersed in a liquid. To prevent particle aggregation, either the pH can be adjusted, or surfactants can be used. Solubility of drug substance particles, however, is often pH dependent; therefore, if the pH is adjusted to provide adequate particle dispersion, the material could dissolve. The same phenomenon can occur with surfactants, where the particles can be solubilized by their addition.

If the particles are not solubilized by the use of surfactants, then for some characterization techniques, the adsorbed layer of surfactant will skew the particle size, making the particles appear larger. This, however, may be relevant to the performance of the drug, and such measurements can be valuable in predicting in vivo performance [2]. Factors such as these must be considered when interpreting physical characterization data for nano materials in pharmaceutical applications.

Particle Size Measurement Via Dynamic Light ScatteringLaser diffraction, the dominant platform for sizing most pharmaceutical powders, is an ensemble technique (i.e. measures many particles rather than a single particle measurement) that has a lower size limit of approximately 0.1 μm. To measure objects below that limit, dynamic light scattering (DLS), also known as photon correlation spectroscopy (PCS), is one of the few ensemble techniques that can be employed in this size range (0.005 – 1 µm). The technique is well established and several commercial platforms are available. The theory and application of this technique are contained in refs. [3-5].

Stable, well-dispersed particles are placed in the sample cell, where Brownian motion causes the particles to move randomly in the suspension. A laser beam passes through the sample cell and is scattered by the particles. The randomly changing diffraction pattern is converted into a histogram of intensity vs. size. Note that this data representation is not what one typically encounters, which is frequency vs. time. For this reason, PCS should be used to measure an average particle size rather than to produce a particle size distribution.

Figure 1 shows a schematic of a particle that has an adsorbed layer of surfactant for particle dispersion. PCS measures a hydrodynamic particle diameter; therefore, the adsorbed layer of surfactant will affect the particle size measurement. The effective particle size is denoted by the circumscribed blue circle. This is not necessarily an undesirable occurrence. If this particle is being designed to pass through a specific

»


PArtIClE sIzIng »

physiological barrier, such as the blood-brain barrier, or certain cell membranes, the hydrodynamic measurement produced by PCS would be extremely relevant. If the purpose of the measurement is to measure the actual size of the particle itself, then the average particle diameter would be larger than observed with other microscopic techniques that would only image the denser particle.

Particle size measurement by Electron microscopySince the inception of nano technology, electron microscopy has served as the gold standard for measuring particle size and morphology. Initially, scanning electron microscopes were incapable of imaging nano-scaled objects; therefore transmission electron microscopy was the only microscopic technique available. With the advent of field emission electron guns for scanning electron microscopes, this scenario changed. Both instruments now offer clear images of nano materials. Figure 2 shows a scanning electron micrograph of drug-carrying biodegradable nanoparticles comprised of a polysebacic acid core and a shell of polyethylene glycol.

Scanning electron microscopy offers a three-dimensional representation of the particles. The gray scale produced in SEM images, however, makes quantitative measurements on these images using appropriate image analysis software very difficult because of the lack of contrast provided.

Figure 3 shows a TEM image of nanoparticles of silicon dioxide (silica) used for drug delivery. Because the electron beam passes directly through the object in a TEM, electron diffraction and fringes can be observed. Rows of atoms can be observed in this image. Additionally, there is much better contract in this image, allowing easier quantification/measurement of features. Even with modern advances in scanning electron microscopy, TEM exhibits much higher resolution.

For both transmission and scanning electron microscopy, the particles must be isolated, and are analyzed in a high vacuum environment. Both techniques are insensitive to adsorbed surface layers such as surfactants that would be required to disperse the particles in a liquid (typically aqueous) environment.

To obtain these high resolution images, in both instances, the sample is bombarded with a very high energy electron beam. Prolonged exposure of the particles to this beam can cause degradation. This must be considered when opting for this type of analysis.

In summary, both PSC and electron microscopy provide information on particle size. If at all possible, both techniques should be employed for the size characterization of pharmaceutical nanoparticles. They are complimentary orthogonal techniques that provide valuable information.

Surface Area MeasurementsParticle diameter is perhaps the most relevant data to collect for scenarios where the physical size of the particle controls the in vivo performance of the drug product. When nano-scaled formulations

Figure 1. Schematic of nano particle with adsorbed surfactant layer. Particle size data obtained by PCS will also measure the

effect of the surfactant [12].

Figure 2. Scanning electron micrograph of drug-eluting nanoparticles [13].:

Figure 3. Transmission electron micrograph of silica particles used for drug delivery [14]

»


« PArtIClE sIzIng

are considered based upon the poor solubility of the drug substance, surface area measurement provides equally meaningful data. It has long been realized that surface area controls the dissolution of solid oral dosage forms [6-11].

Because nanoparticles can easily pass through the filters and frits found in commercial surface area instruments, either the sample must be exposed to vacuum very slowly, or the material should be presented in an agglomerated form (i.e. nano-scaled). If the nanoparticles are contained in aggregates, a two-tiered strategy for characterization could be adopted, whereby the particle size measurement would measure aggregate size, and surface area measurements would provide an indication of the nano particle size. Often, high energy sonication is employed in an attempt to attain the primary particle size. This is not a preferred choice. The data obtained is often an indication of the degree of sonication, rather than the primary particle size.

Isoelectric Point and Zeta PotentialIsoelectric point and zeta potential measurements are routinely collected data by practitioners working with inorganic sub-micron powders, nano particles, or colloidal systems. The origin of these measurements is found in the pioneering work performed by Stern to describe the interaction of particles in liquids. For nano particles such as proteins, liposomes and nano crystals with low isoelectric points, i.e. with low surface charges, the nano material will easily flocculate in the suspension, creating an undesirable scenario.

For particles with a high zeta potential (either positive or negative), all of the particles will have a high surface charge, thereby repelling each other in the liquid and maintaining a discrete identity. The isoelectric point of a particle in a liquid is the pH at which the surface charge on the particle is zero, which is usually avoided in pharmaceutical nanoparticulate applications. (For waste water treatment, the water may be pH adjusted to achieve the isoelectric point, thereby causing harmful particulates to flocculate and allow them to be removed via filtration). Figure 4 shows the classic diagram used to describe the layers that form around a particle in a liquid

There are two analytical instrument platforms used to measure zeta potential: electrophoretic and electroacoustic. With electrophoresis, suspended particles are placed between the plates of a capacitor, and the potential across the plates is scanned (typically from -25 to 25 mV). The particle motion is measured as a function of potential using many of the same principles employed in dynamic light scattering measurements. (Instruments are available that perform both particle size measurement via DLS and zeta potential measurements.) With electrophoretic measurements, the particles must be small enough to remain suspended without the introduction of any additional energy or agitation (stirring, pumping, sonication, etc.) Additionally, the suspension must be suitably dilute to permit individual particle motion detection by the laser.

Figure 4. Diagram illustrating particle surface charge in a liquid as a function of distance from the surface of the particle [15].

»


PArtIClE sIzIng »

Electroacoustic instruments require more concentrated suspensions, and the suspension can be stirred during the measurement. With this technique, ultrasonic bursts of energy are pulsed through a suspension, causing charged particles to move. This motion generates an electrical charge, which is monitored as a function of the ultrasonic energy burst. The sample can be stirred because the duration of the pulse is so small; particle motion created by stirring is negligible. Highly concentrated suspensions (up to 50 vol. %) can be measured used this technique.

If the zeta potential of the nano material in its nascent state is insufficient to prevent flocculation, pH adjustments can be made or surfactants can be added. The same limitations and cautions described in Sample Preparation, however, are applicable here; pH changes and the addition of surfactants can easily solubilize the nano material.

The need for a rigorous Physical Control strategyThe mandate for any commercial drug product is to deliver safe and efficacious treatment for human illness. All pharmaceutical companies are required, by law, to demonstrate that they understand the therapeutic action of the medicine, and they can reproducibly manufacture the material to the same (or better) level of quality over time. Physical and chemical tests are performed in a rigorous fashion before any medicine can be released for human use. Such tests include chemical tests for purity and potency, dissolution behavior, moisture content; and often physical tests such as particle size (laser diffraction, sieve testing, etc.) and perhaps specific surface area. For drug product performance, typically dissolution testing is heavily relied upon because this testing most closely imitates how the product will perform in vivo, provided the drug product manufacturing process is robust.

For nano materials in the pharmaceutical industry, the need for this control strategy is even greater, specifically for the physical properties of the drug substance. The performance of the medicine is directly linked to these properties. These properties will maintain themselves only if the formulation is shown to be stable over the shelf life of the drug product. It is quite likely traditional CMC strategies will be augmented or changed to meet the more demanding needs of nano medicines, and more sophisticated physical tests will be required in the development of these compounds. Some of these techniques may also be used as quality control tests to release the medicine prior to human consumption.

This additional testing may require pharmaceutical companies to develop physical reference stands for nano medicines, much in the same way reference standards are used for potency and related substance testing today. Because drug product performance is linked to the drug substance particle size in the formulation, reference standards may be required for some physical property testing.

summaryNano medicines offer unique therapeutic advantages over their traditional counterparts by exhibiting unique properties such as enhanced solubility, selective cell targeting, and the ability to pass certain biological barriers and membranes. This enhanced performance/capability is linked directly to the physical properties of the material. No single test can provide an adequate description of the material. Great care must be taken to ensure that the sample preparation steps do not alter the material. Tests not used routinely in the pharmaceutical industry may provide the appropriate level of scrutiny to interrogate the material. It may become necessary to use some of these new techniques for quality control testing. Reference materials could be necessary to ensure that physical testing data are consistent and reliable for a given nano material.

references1. Pathak, Y. and D. Thassu, eds. Drug Delivery Nanoparticles

Formulation and Characterization. 1 ed. Drugs and the Pharmaceutical Sciences, ed. J. Swarbrick. Vol. 191. 2009, Informa Healthcare: London. 394.

2. Banker, G.S. and C.T. Rhodes, eds. Modern Pharaceutics. 4 ed. Drugs and the Pharmaceutical Sciences, ed. J. Swarbrick. Vol. 121. 2002, Marcek Dekker: London. 838.

3. Xu, R., Particle Characterization: Light Scattering Methods. 2000, Dordrecht: Kluwer Academis Publishers. 397.

4. Merkus, H., Partucle Size Measurements: Fundamentals, Practice Quality. Particle Technology Series. Vol. 17. 2009, New York: Springer. 532.

5. ISO, 13321-1: Particle Size Analysis -- Photon Correleation Spectroscopy. 1996, International Standards Organization: Geneva.

6. Iacocca, R.G., Physical Characterization Tests for API used in Low Dose Formulations, in Formulation and analytical development for low-dose oral drug products, J. Zheng, Editor. 2009, Wiley & Sons: New York. p. 309-324.

7. Iacocca, R.G., C.L. Burcham, and L. Hilden, Particle engineering: A strategy for drug substance physical property control during small molecule development. J Pharm Sci, 2010. 99(1): p. 51-75.

8. Fukunaka, T., et al., Dissolution characteristics of cylindrical particles and tablets. International Journal of Pharmaceutics, 2006. 310(1-2): p. 146.

9. Horter, D. and J.B. Dressman, Influence of physicochemical properties on dissolution of drugs in the gastrointestinal tract. Advanced Drug Delivery Reviews, 1997. 25(1): p. 3-14.

10. Thomas Schreiner, U.F.S.H.L., Immediate drug release from solid oral dosage forms. Journal of Pharmaceutical Sciences, 2005. 94(1): p. 120-133.

11. Jinno, J.-i., et al., Effect of particle size reduction on dissolution and oral absorption of a poorly water-soluble drug, cilostazol, in beagle dogs. Journal of Controlled Release, 2006. 111(1-2): p. 56.

12. courtesy of D. Shekhawat

13. http://www.futurity.org/health-medicine/drug-toting-mucus-busting-nanoparticles, Ben Tang and Mark Koontz/Johns Hopkins University

14. http://en.wikipedia.org/wiki/File:Mesopourus_silica_closeup.jpg

15. http://en.wikipedia.org/wiki/Double_layer_(interfacial)

»


« PArtIClE sIzIng

Author BiographyRonald Iacocca received his B. S., M.S. and Ph.D. in Materials Engineering from Rensselaer Polytechnic Institute (RPI). For nine years prior to joining Lilly, he was a faculty member in the Department of Engineering Science and Mechanics at The Pennsylvania State University. In November 2000, Ron joined Eli Lilly as a research scientist in Physical and Structural Characterization, Pharmaceutical Product Research and Development. Currently, he is a Senior Research Advisor in Analytical Sciences Research and Development, and is team leader of the Materials Science/Physical Characterization team. He has authored/co-authored over 65 journal articles, book chapters, and review articles, and has been

granted 4 patents. He is a member of ASM International, ASTM, and ISO. In 2005, he was selected as one of ninety world-wide experts by the Bill and Melinda Gates Foundation to participate in the road-mapping initiative for the development of a world-wide malaria vaccine.

Dr. Iacocca also served as an adjunct professor in the Department of Industrial and Physical Pharmacy at Purdue University from 2006-2009, and serves as a member of the editorial advisory board for American Pharmaceutical Review, as a reviewer for the Journal of Pharmaceutical Sciences, as a key reader for Metallurgical and Materials Transactions A, and is head of the working group on laser diffraction for the International Standards Organization (Working Group 6, ISO TC24/SC4). In 2010, he was elected to the USP to serve on the Physical Analysis Expert Committee

Many pharmaceuticals, such as monoclonal antibodies are produced from genetically engineered mammalian cell lines. Purity with regard to undesired host-cell proteins (HCP) of final product is essential for product safety. Process development by varying upstream conditions and analyze the effects on downstream processing is needed for optimal yield and purity. We have characterized HCP patterns in purification of monoclonal antibodies from CHO cells by using 2-D Fluorescence Difference Gel Electrophoresis (2-D DIGE).

2-D DIGE technology with two different samples and a pooled internal standard per gel pre-labeled with CyDye™ DIGE Fluor minimal Dyes, detects differences in protein abundance. Experimental variation is virtually eliminated and the quantitative data is very reliable.

Differences in protein expression between culture supernatants grown with a set of altered media compositions were analyzed. Also, differences in the HCP patterns of MabSelectSuRe™ eluent fractions were analyzed. The results were related to yield of target protein and HCP levels obtained with ELISA assay.

Presents this interactive webcast

Characterization of host cell protein patterns using 2-D DIGE

Monday, March 21 3pm CET /10am EDT

Register online at:http://tinyurl.com/4bo7usx

Speaker:

Maria Winkvist, Ph.D.GE Healthcare, Life Sciences, Sweden

Maria Winkvist trained as a cellular and molecular

biologist at The Ludwig Institute for cancer research in

Uppsala, Uppsala University in Sweden, where she

received a Ph.D. in Medicine 2007. Her research was

focused on transcription regulation and cell signalling.

In 2007, Maria Winkvist joined GE Healthcare, Sweden,

as a specialist for Biomolecular imaging. She is part of

the Scientific Support group, responsible among other

things for application development and training both

internally and externally.

Moderated by: Rita Marouga

»


rAmAn »

Joanny Salvas1, Jean-Sébastien Simard1, Ryan Gosselin, Ph.D.2 & Nicolas Abatzoglou, Ph.D.2 1 Process Analytical Science Group, Pfizer Montréal2 Université de Sherbrooke, Department of Chemical & Biotechnological Engineering, Industrial Research Chair Pfizer/UdeS on PAT in Pharmaceutical EngineeringE-mail: [email protected]

IntroductionContextThe use of PAT has spread throughout the pharmaceutical industry; mainly because PAT makes it possible to gain useful process insight, improve monitoring throughout the manufacturing steps, lower costs associated with the use of wet-chemistry-based testing methods, and increase overall efficiency. They are tools that fit very well in the QbD scheme for better process control and understanding.

Many of the PAT methods are based on MultiVariate Predictive Models (MVPM), and the calibration of such model is a step crucial to the success of the MVPM-based PAT method.

The current trends in MVPM calibration are aligned with the mantra “the more the better”. This implies that using more calibration points, with more samples, manufactured as closely as possible to the commercial product, with more precise reference methods, will lead to more precise, better performing MVPM-based PAT methods. Obviously, it also leads to a costlier and more time consuming development process, thus limiting the potential applications as well as lowering the overall payback.

Available literature, to the authors’ knowledge, does not contain studies in which the predictive ability of multivariate models elaborated with various calibration approaches is evaluated as function of the latter. This is true for both Raman spectroscopy and many other spectroscopic techniques. Thus, this work has been undertaken to understand the influence that different calibration parameters have on the final performance of predictive multivariate models and contribute to optimize the calibration process to minimize cost and maximize performances.

This study was conducted using data collected during an actual PAT application development. Salvas et al [(3),(4)] reported on the development of a PAT for the quantification of minerals in intact tablets, using Raman spectroscopy. The development of this project created the opportunity to study more thoroughly the calibration processes of multivariate models. From this application, out of the four minerals for which multivariate models were developed, two were selected to be part of the investigation: one in high concentration and one in low concentration. They both come from the same commercial product. They are referred to as Mineral 1 and Mineral 2 throughout this article.

Calibration of Multivariate Predictive Models: The Study of Factors Influencing the Prediction Accuracy of Raman Spectroscopy Applied to Pharmaceutical Tablets

»


« rAmAn

scope and methodologyThe development of an MVPM includes several critical activities, one crucial being the sample preparation for model calibration. Common practice dictates that samples must be prepared in such a way that they are as close as possible to “real” samples (i.e. taken from the production line). To insure this similarity with full-scale situations, scaled-down lab- and pilot-equipment are often used. While this makes it possible to obtain more representative calibration samples that provide good coverage of the calibration span, it is a very time-consuming and costly activity.

As suggested by the QbD approach for developing a new product – or a new method, in the present case – several parameters were selected to be part of the present study [(5)]. The monitored response for each trial was taken to be the accuracy of the final method.

Eight factors (parameters) were identified and tested independently on two data sets. The tests were guided by a design of experiments (DoE), making it possible to optimize the total number of predictive models that needed to be made, while retaining sufficient data to allow minimal confounding within second-order factorial interactions. Each run of the DoE represent a multivariate model that is calibrated with data obtained with a different set of sample-manufacturing or

data-organizing parameters. Factors and their levels are detailed in Table 1.

In this work, a batch of sample refers to a given set of samples all manufactured under the same conditions and at the same time, corresponding to one calibration point (one set of materials concentration). This work required the manufacturing of 26 batches for 13 calibration points with each 2 manufacturing protocols (different tablet presses, same overall batch volume). Replicates refer to individual samples taken from the same batch.

Table 1: Parameters and tested levels for the DoE

PARAMETER (alternate notations) LEVEL TESTED

Type of press used for sample manufacturing (Factor A, aka Press)

Automatic (27-station press)Manual (single-punch press)

Distribution of calibration points over the calibration span (Factor B, aka Distrib.)

Evenly distributedConcentrated around target

Number of calibration points (Factor C, aka NbPts)

Low (half )High (all)

Number of replicates within each calibration point (Factor F, aka NbRep.)

Low (half)High (all)

Use of commercial samples for model calibration (Factor E, aka Prod.)

PresentAbsent

Calibration algorithm used (Factor D, aka Algo)

PCRPLS

Reference data used for modeling (Factor G, aka Meas.)

Reference method value Experimental manufacturing values

»


rAmAn »

A 27-3 design obtained with generators E=ABC, F=BCD and G=ACD allowed to obtain level IV resolution, leading to double interactions being confounding with each other but not with the main effects. Another useful particularity of this fractional design is that it can be “collapse[d] into either a full factorial or a fractional factorial in [many subsets] […] of the original factors”. In the event that 3 factors are found to be non significant, the probability of being able to transform the design in a full 24 design is high [(2)].

Samples and calibration datasets were prepared according to the DoE. Specifics regarding sample manufacturing and model preparation are reported in a previous paper and master thesis: Salvas et al [(3), (4)]. These details are not repeated here and are not mandatory for interpretation of the present study.

results and DiscussionSummarized results obtained after execution of the DoE are presented in Table 2. The response monitored for the ANOVA is the accuracy of the models and it is expressed as a form of error: the absolute value of the relative difference between the Raman and the reference values. Reference values were obtained by Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES).

All following analyses are based on the results presented in Table 2.

Effects of individual factors were determined through two separate analyses of variance (ANOVA): one for each Mineral dataset. Results of ANOVA tests are summarized in a modified Pareto chart, illustrated in Figure 1. In this figure, a bar that continues past the significance

limit (p = 0.05, dashed line) indicates that the associated factor, in the associated ANOVA, was found to have a significant effect on the response. For example, factor B (Distribution of calibration points) is demonstrated to have a significant effect in the case of the ANOVA on Mineral 2, but not on Mineral 1.

Figure 1: Modified Pareto chart of ANOVA – main effects

Figure 2: Plot of means – Mineral 1

Figure 3: Plot of means – Mineral 2

Table 2: Prediction results for DoE models

RUN

Pres

s us

ed

Poin

t di

stri

butio

n

Nb

of

calib

ratio

n pt

s

Alg

orith

m

Com

mer

cial

sa

mpl

es

Nb

of re

plic

ates

Refe

renc

e da

ta | RELATIVE DIFFERENCE |

MINERAL 1 MINERAL 2

% %

1 -1 -1 -1 -1 -1 -1 -1 7.34 7.82

2 +1 -1 -1 -1 +1 -1 +1 6.85 1.64

3 -1 +1 -1 -1 +1 +1 -1 4.23 6.92

4 +1 +1 -1 -1 -1 +1 +1 2.34 9.24

5 -1 -1 +1 -1 +1 +1 +1 4.13 55.93

6 +1 -1 +1 -1 -1 +1 -1 1.28 30.24

7 -1 +1 +1 -1 -1 -1 +1 3.75 45.78

8 +1 +1 +1 -1 +1 -1 -1 4.85 38.22

9 -1 -1 -1 +1 -1 +1 +1 8.27 6.09

10 +1 -1 -1 +1 +1 +1 -1 7.98 11.44

11 -1 +1 -1 +1 +1 -1 +1 2.55 6.36

12 +1 +1 -1 +1 -1 -1 -1 9.13 1.78

13 -1 -1 +1 +1 +1 -1 -1 1.63 79.23

14 +1 -1 +1 +1 -1 -1 +1 1.14 60.52

15 -1 +1 +1 +1 -1 +1 -1 1.79 46.99

16 +1 +1 +1 +1 +1 +1 +1 3.19 21.45

»


« rAmAn

significant Parameters: Press used (A)Test results indicate that factor A has a significant effect on the measured response in the case of Mineral 2 (low concentration), but not with the Mineral 1 dataset.

The use of a single-punch manual press requires small-scale repetitive movements, such as measuring a few grams of the mixed powder and letting it slide in the compression matrix. Despite using mitigating techniques (such as limiting vibrations and transfers), the smaller-scale movements and manipulations apparently still induced more local heterogeneity than bigger-scale automatic presses. These local heterogeneities seem to be more detrimental when trying to link bulk concentrations to sub-samples of less-concentrated materials. It might be that increasing sampling volume when acquiring the Raman spectra could help mitigating this effect, but this remains to be tested.

significant Parameters: Distribution and number of Calibration Points (B and C)Factor B is significant only for the high concentration material (Mineral 1), while Factor C is significant for both. Since some inconsistencies in the data have been observed, several additional tests and analyses were done regarding these two factors (details available in (4)).

These additional tests have led to the hypothesis that an uncontrolled parameter was most probably affecting the response. Several attempts at pinpointing this parameter finally lead to the conclusion that it was the population in the subsets of points selected for a given run that affected the response. Indeed, when the number of calibration points used is the maximum available (such as in run 5 for example), there is no positive degree of liberty left when assembling the calibration matrix: all sample batches are used. However, if the required number of point is lower than the maximum available, a choice must be made regarding which calibration batches are selected for model elaboration.

During this work, when establishing a sub-dataset, the line of thought has been to 1) cover the whole range and 2) include batches in such manner as to have an even distribution of points

along the span. Thus, calibration matrices were prepared for each run, under these constraints. But, with lower levels, several alternatives could satisfy the constraints. For example, dataset containing mixes 3, 8, 10, 11 & 13 or mixes 3, 4, 7, 8 & 12 both satisfied the constraints. Given this situation, and the fact that problems with some batches had been spotted previously [(3), (4)], it was advisable to study the effect of this choice on the response.

A new design of experiment was prepared in order to infirm or confirm the hypothesis that, when using the same number of calibration

WITec GmbH, Ulm, Germanyphone +49 (0)731 140700, [email protected] www.witec.de

Confocal . Raman . Fluorescence . AFM . SNOM

WITec´s new True Surface Microscopy allows confocal Raman imaging guided by surface topography. The topographic coordinates obtained by an integrated profi lometer are used to perfectly follow the sample surface in confocal Raman imaging mode.

The result is an image revealing chemical properties at the surface of the sample, even if it is rough or inclined.

Automated Raman-AFM System alpha500 with Attached Sensor for Profi lometry

Topographic Raman Image of a Pharmaceutical Tablet

True Surface Microscopy

Topographic Raman Imaging

NEWPRODUCT

WITec Instruments Corp.phone +1 865 984 4445, [email protected]


» rAmAn »

points, changing the members included in the selection had an effect on the response. Several subsets of 5 batches (i.e. calibration points) were prepared and used to obtain models, with an equal number of replicates in each batch.

Figure 2 and Figure 3 illustrate that, depending on the group of points selected and for both components studied, there is a significant effect on the response.

Under the light of this new data, it has become clear that the influence of factors B and C (distribution and number of calibration points) can be equally due to the selection of the points amongst all that were available as well as the added number of batches themselves. This could mean that, since it is impossible to foresee which batch will turn out to contain “good” variance and thus improve the model calibration, disposing of a high number of calibration points from which to choose an adequate sub-set for model calibration might reveal to be more of an advantage than saving costs by reducing the number of available different batches. The optimal level for factor C must hence be high, so the best selection can be made amongst available batches, no matter if the points are equally distributed or not.

non significant ParametersWithin the framework of this study, the following factors do not appear to have significant effect on the response, for neither the Mineral 1 nor the Mineral 2 datasets:

• factor D (calibration algorithm used);

• factor E (use of commercial samples in the calibration set);

• factor F (number of replicates used per calibration point); and

• factor G (type of reference measurement used in the calibration step).

The conclusion reached for factor D is in accordance with literature on the subject. It is indeed widely agreed that both PLS and PCR algorithms can lead to a good model, but that, in general, PLS will do it using fewer Principal Components (PC) [(1)].

No literature was found on the use of commercial samples in the calibration stage of an MVPM-based PAT (factor E), but the results obtained are in accordance with the general calibration procedure. As the goal is to produce samples that are as close as possible to the future ones (commercial), it is not surprising that, when such goal is achieved, including the real commercial samples in the calibration set does not make a statistically significant difference. In the case real scale samples are difficult to obtain, the inclusion of commercial samples in the calibration set might be an asset for fine-tuning the model, but this remains to be tested, as it was not part of the present work.

The conclusions reached in the case of factor F are a pleasant surprise. It was expected, in accordance with the general calibration procedure, that including more replicates, and thus more variation in the calibration matrix, would allow the model to better perform in both the validation stage and its routine use. It appears that, no matter how many replicates are included in each calibration point, the response is not affected. It was expected that a decreasing asymptote-like response-plot of prediction accuracy vs replicate number would be obtained when increasing the number of replicates This would have suggested an optimal number of replicates at the lowest error obtained with the least number of samples; it was not the case. The results obtained can be explained by the fact that the commercial samples are manufactured in such a manner that they are very uniform from batch to batch; thus, the inclusion of more variation in the calibration stage may not be as crucial as in other cases where more variation will be encountered in the application of the model. On the other hand, including more replicates in the calibration stage may increase model robustness over time. This was not investigated in the present study and should be kept in mind in future work.

Enwave

Raman Solutions forPAT Success!

On-line Process

Laboratory Discovery

�ul�ill �our anl�tical goals with better tools

Enwave Optronics, Inc.Tel: (949) 955-0258

[email protected]

Handheld Raman

ProRaman-I

ProRaman-L

Raw Material Release

Optronics

»


« rAmAn

The results obtained regarding factor G are the most surprising. It was expected that a significant difference would be observed when using reference methods rather than theoretical values for y-variables, especially for low-concentration materials such as Mineral 2. This was expected due to (1) the “poorer quality” of the samples developed at lab-scale, (2) the low concentration values of the raw materials and (3) the sub-sampling inherencies of spectroscopic techniques. Indeed, because Mineral 2 is less concentrated, a slight error in the manipulations should have had a higher impact on the accuracy of the expected values associated with the samples. Moreover, the higher variation of the sample-to-sample inhomogeneities in the probed-region was expected to cause even more inaccuracies regarding the expected concentration-values of each individual tablet and spectra. Alternatively, a mineral with higher concentration was expected to be distributed more homogeneously in each sample, thus decreasing the impact of potential manipulation errors on the accuracy of the theoretical values associated with the batch.

In light of the results obtained, the previous explanations must be revisited. First, it can be noted that the reference measurements have an error (or precision) of their own. Including the reference measurements in the calibration stage includes their error in the model. Theoretical values also have an error, but there is no a priori knowledge as to which error is “better”, even though it was originally

postulated that the reference acquired with a compendial method should de facto be. Training the model with the reference error was expected to systematically give better result.

The mean difference between the theoretical and the reference values was checked for the samples manufactured with the automatic press, and it is of 1.9 and 1.2 % for Mineral 1 and 2, respectively. This deviation is within the reference method’s precision range, suggesting that the difference might not be statistically significant. Under the light of this information, the conclusions reached regarding the effect of factor G make more sense. Both concentration measurements are apparently equivalent, showing that the models are equally well-calibrated. The difference between reference and theoretical values are greater when dealing with samples manufactured using the manual press (respectively 3.9 and 2.3 %), but still not big enough to have an impact on the accuracy of the multivariate model. The difference is nonetheless in agreement with the fact that factor A (press used) was found to have a significant effect on the response.

Nevertheless, since manipulation errors are intrinsically various and mostly not repeatable, it is possible that, while in this case they are equivalent to that of the reference method, they might be higher or lower in any other case. More work should hence be done on that subject.

Raman solutionsfor the pharmaceutical industry

email: [email protected] www.pharmaraman.com

Sometimes you have to look below the surface.

From fast imaging tools in drug discovery throughtargeting bottlenecks in drug development and providing new technology solutions for assuring drug product quality in manufacturing, HORIBA Scientifi cis continuously meeting new challengesacross the breadth of the industry.

Raman solutions- iceberg half page for APR.indd 1 2/4/2011 12:18:40 PM

»


rAmAn »

optimized Protocol ProposalConclusions reached regarding the effect of each factor on the accuracy of the method allowed proposing an optimized development protocol for MVPM-based PAT methods, as described in Table 3.

An automatic press should be used to produce a high number of evenly distributed calibration points. A PLS regression algorithm should be used to calibrate the predictive model, using theoretical y-variables as a starting point. The number of replicates can be lowered significantly if it makes it possible to gain time or reduce efforts and/or costs.

Using these sets of parameter settings should allow the development of a reliable predictive model, at reduced cost and with an accelerated schedule. This optimized method was tested in order to verify the overall impact on performance by selecting the data in the calibration matrix that would have been obtained if such a protocol had been used. Three (3) more multivariate models were hence developed, one for each of the monitorable minerals presented in Salvas et al [(3), (4)]. Prediction errors, calculated as the relative difference between the claim and the measurement, are presented in Table 4. The difference in precision does not exceed 2.5 %(absolute value) in the worst case observed and would have allowed savings in direct invested man-hours of approximately 30 % and overall development-cost, by 31 %.

ConclusionThis work has resulted in the recommendation of an optimized development protocol for Process Analytical Technology based on MultiVariate Predictive Models (MVPM-based PAT).

Beside the monetary savings the main advantage of these two changes is that it allows cutting the estimated development time by two. This is a tremendous advantage, given the always limited resources that can be allocated to such projects. This time reduction allows the facility to undertake more different projects at the same time and hence multiply the potential gains and return on each system investment.

This work was based on a specific PAT application (Raman spectroscopy for content-monitoring of intact tablets). Further work is undergoing with different applications (other spectroscopic techniques, other types of products, other parameters) in order to pursue and challenge these conclusions. Many other parameters could affect method performance, but the present work has demonstrated that it is possible and wishable to lower the burden of multivariate model calibration by optimizing the development protocol.

AcknowledgmentsThe authors are indebted to Pfizer Montreal, The National Science & Engineering Research Council (NSERC) of Canada and the Université de Sherbrooke for funding related with the project.

Anne-Marie Demers and the late Jasmin Groleau are thanked for their precious help in preparing the many samples required for this work.

Ryan Gosselin and Guillaume Leonard are also kindly thanked for their help in revising this article.

references1. Esbensen, K.H. (2004.) Multivariate data analysis – in practice. 5th edition, Esbjerg, Camo

Process AS, 598 p.

2. Montgomery, D.C (2001). Design and Analysis of Experiments. 5th edition, Wiley, New York, 684 p.

3. Salvas, J., Simard, J.-S., Abatzoglou, N. (2010a) Raman Spectroscopy to Analyze Intact Pharmaceutical Tablets, American Pharmaceutical Review, April 2010, p. 46-53

4. Salvas, J. (2010b). Calibration of multivariate predictive models: the study of factors influencing the prediction ability of Raman spectroscopy applied to pharmaceutical tablets, Master thesis, University of Sherbrooke, Canada.

5. United States. International Conference on Harmonisation (2008). ICH Harmonised Tripartite Guideline – Pharmaceutical Development – Q8(R1). Current Step 4 version dated 13 November 2008, accessed online (http://www.ich.org/LOB/media/MEDIA4986.pdf) September 01 2009, 28 p.

Table 3: Recommended levels for each tested factor

FACTORLEVEL

TESTED OPTIMAL RECOMMENDED

(A) Type of press used Automatic, Manual Automatic Automatic

(B) Distribution of calibration points Concentrated, Even Even Even

(C) Number of calibration points Ca: 3, 5, 6, 8, 12, 13 Mg: 4, 5, 6, 8, 9

High number

High number

(D) Calibration algorithm used PCR, PLS - PLS

(E) Use of commercial samples Absent, Present - Most convenient

(F) Number of replicates 3, 5, 7, 10 - 5

(G) Type of reference used Theoretical, Reference - Theoretical

Table 4: Comparison of error for optimized models

MODELS MINERAL 1 MINERAL 2 MINERAL 3

FULL MINIMAL FULL MINIMAL FULL MINIMAL

% % % % % %

Prediction error 2.9 1.9 2.6 3.1 5.6 8

Differencefull vs minimal

-1.0 +0.5 +2.4


« rAmAn »

BiographyDr. Nicolas Abatzoglou is full professor and Chairman of the Department of Chemical & Biotechnological Engineering of the Université de Sherbrooke (UdeS). He is a specialist in Process Engineering involving particulate systems in reactive and non-reactive environments. He is the holder of the UdeS’ Pfizer Industrial Research Chair on Process Analytical Technologies (PAT) in Pharmaceutical Engineering. He is co-founder of the company Enerkem Technologies Inc., a spin-off of the Université de Sherbrooke in the field of energy from renewable resources. He has a career of many years at both the academic and industrial levels. His professional experience as engineer spreads over the last twenty years. He represented Canada at the International Energy Agency (Gasification Task) from 1997-2001 and was the secretary of the Board of Directors and the Executive Committee of the AQME (Association québécoise pour la maîtrise de l’énergie) from 1996-2000. His production as a researcher includes a hundred of publications in scientific reviews, international conferences, plenary and invited lectures, patents and a book chapter.

Jean-Sébastien Simard has a Master degree in Chemical Engineering specialized in particulate systems for direct compression from Université de Sherbrooke, Québec, Canada. He is currently pursuing a MBA degree at Université Laval, Québec, Canada. He has been with Pfizer Canada for the last ten years, where he worked as a Product and Process Development Scientist for the pharmaceutical processing unit. For the last five years, he has been responsible for the Process Analytical Technology Development Group in the Technical Services. He has co-authored many different presentations on particulate systems behavior, Quality by Design and Process Analytical Technology applications. He is also the Industrial Responsible of the UdeS/Pfizer Industrial Research Chair on Process Analytical Technologies in Pharmaceutical Engineering.

Joanny Salvas has a bachelor’s degree in biotechnological engineering. She has completed a M.Sc.A. in chemical engineering with the Université de Sherbrooke, conducting her graduate work at Pfizer Montreal’s facility (Canada), as part of a Chair partnership. Her research aimed at optimizing the development protocol of multivariate predictive models used as part of PAT methods. She currently is a PAT Scientist at Pfizer Montreal, where she pursues several projects with different technologies such as Raman spectroscopy and Rapid Microbiological Methods. She is also leading projects for international sites.

Dr. Ryan Gosselin is an assistant professor at the Department of Chemical & Biotechnological Engineering of the Université de Sherbrooke, Canada. He is a specialist in Process Engineering and on-line quality control through the use of multivariate data analysis and chemometrics. As a member of the Pfizer Industrial Research Chair on Process Analytical Technologies (PAT) in Pharmaceutical Engineering, his present work focuses mainly on issues relating to the production and handling of non-reactive particulate systems.

Don’t Burn Through Your MoneyBurn Through Your Work!Our DSC Instrument features: User-replaceable/corrosion resistant sensor to minimize maintenance costs TOPEM, the new multi-frequency modulated temperature DSC technique from METTLER TOLEDO High heating rates to reduce measurement time and increase productivity Reliable auto-sampler to maximize sample analysis and throughput

www.mt.com/thermalexcellence

»


DIssolutIon »

Saeed A. QureshiSenior Research Scientist, Therapeutic Products Directorate,Health Canada, Banting Research Centre Email: [email protected]

Limitations of Some Commonly Described Practices in Drug Dissolution Testing and Suggestions to Address These

IntroductionDissolution tests are employed to establish the quality of drug products, mostly tablets and capsules, based on in vitro drug release characteristics of these products. In reality, a dissolution test may be considered as a simple extraction step in a vessel with a stirrer. Most of the commonly used apparatuses in this regard are known as paddle and basket apparatuses, in which a round bottom vessel (1 L) containing a stirrer referred to as paddle (an inverted T-shaped bar) or small wired cage (known as basket), respectively, are used. These apparatuses are very well recognized and used around the world with the acceptance of regulatory and standard setting organizations. Detailed descriptions about these apparatuses may be found in any of the most commonly followed pharmacopeias such as United States Pharmacopeia (USP) [1].

As noted above, drug dissolution testing is a relatively simple technique, however, serious concerns and problems are often reported in the literature about it [2-5]. These reported problems often relate to: (1) failing of the performance evaluations of the apparatuses (calibration) and/or products; (2) lack of establishing the link between in vitro dissolution results and in vivo results, commonly referred to as in vitro-in vivo correlations or IVIVC; (3) lack of objectivity in setting or selecting experimental conditions for product evaluations (4) setting unreasonably wide tolerances based on complex and convoluted rationales. These wide spread concerns result in frustrations, within both regulatory and manufacturing environments, where objectivity and reliability of an analytical technique is of critical importance for establishing the standards for the assessment of quality of the drug products.

With such frustrations, it has been suggested that the dependence on drug dissolution testing should be eliminated [6]. As drug dissolution testing is an important and relevant step, the question obviously should be that what went wrong in the practice of drug dissolution testing rather than removal of the test that is mandatory [7]. This article will present a discussion as to why there are such concerns and describe some solutions to address these concerns.

»


DIssolutIon »

Background Information – objective of Drug Dissolution testing The quality of an oral drug product (tablet and capsule) is based on the fact that the drug will be released from a product in a predictable and reproducible manner and dissolved in the fluid present in the human gastrointestinal (GI) tract, in particular, small intestine. Thus, this in vivo drug dissolution step, also interchangeably referred to as drug release, becomes a critical step for developing a product and later assessing its quality.

A drug dissolution test, or simply dissolution test, is conducted to mimic the above described in vivo dissolution behavior of the drug in vitro. It cannot be emphasized enough to highlight the fact that a drug dissolution test is a test to evaluate in vivo dissolution behavior of a drug product. There is no other objective or rationale for conducting this test. However, there is a common practice for describing and using the dissolution test, without its stated link to in vivo, for establishing batch to batch consistency of the product and this is referred to as a quality control (QC) test.

Unfortunately, this appears to be a misconception about the practice of drug dissolution testing and leads to current problems and concerns about the technique. If the link to the in vivo behavior is ignored, then the obvious question would be: what parameter/characteristic, or consistency thereof, a dissolution test reflects. In addition, what would be the basis of selecting experimental conditions to conduct a dissolution test? It is, therefore, important and critical to note that the only purpose or objective of dissolution testing is to assess the in vivo release behavior of a product. Keeping this objective in mind should also help and guide in defining experimental conditions for dissolution testing.

Evaluating and relating to In Vivo Dissolution BehaviorOnce the objective is set, as described in the previous section, then the question should be how one would relate the in vitro results to in vivo dissolution characteristics? This question should be answered in two parts. The first part addresses the fact that the dissolution test be conducted by mimicking, not necessarily duplicating, the in vivo or intestinal environment. The second aspect should be a comparison of the in vitro results to the in vivo. The discussion regarding the first answer is provided in the following section, however, discussion on the second aspect is provided in this section.

In this regard, the most commonly reported practice is that of developing or establishing an in vitro-in vivo co-relationship or IVIVC. The commonly cited definition of IVIVC from the US FDA guidance document is: “It defines IVIVC as a predictive mathematical model describing the relationship between an in vitro property of a dosage form (usually the rate or extent of drug dissolution or release) and a relevant in vivo response, e.g., plasma drug concentration or amount of drug absorbed” [8]. The preferred or desired IVIVC outcome is of

level “A” which implies comparing point-by-point (time-by-time) in vitro dissolution results with in vivo dissolution results extracted from drug concentration-time profiles. Conversely, by comparing predicted drug concentration-time profiles obtained from in vitro dissolution results with the actual drug concentration-times profiles obtained from bioavailability/bioequivalence (BA/BE) studies. Apart from lack of clarity on the mechanics (procedure) of obtaining in vivo dissolution results or deriving blood levels from in vitro dissolution results, suggested IVIVC practices appear to have serious limitations. For example: As the name IVIVC implies that one is required to develop relationships between in vitro and in vivo results. However, in practice, conducting a dissolution test is never meant for establishing such a relationship as this relationship is considered to be always present. In fact, existence of this relationship (IVIVC) forms the basis of conducting of a dissolution test. It appears that there is serious confusion in the literature in this regard. The purpose of dissolution testing should be or has always been to evaluate characteristics (quality) of the product, based on the underlying principle that a dissolution test relates well to the product’s in vivo dissolution characteristics.

Even when such a relationship is developed, as current practices require, by conducting both in vitro (dissolution) and in vivo BA/BE studies using single product or multiple products with different formulation/manufacturing variations, the question becomes what did one achieve from this practice. If one gets a perfect correlation then it would show that dissolution results are capable of predicting in vivo results. Is it not that a dissolution test is conducted based on this principle in the first place, i.e., a dissolution test is conducted to reflect potential in vivo behavior of a drug product. Then, why does the development of IVIVC have to be repeated with every drug and product?

There is another major flaw in the current practices of IVIVC. These practices of so called IVIVC seek a matching (rather than relationship) by adjusting experimental conditions so that in vitro results would match the in vivo outcome. Thus, in reality, the practice of IVIVC has become a practice of searching test/experimental conditions to match in vitro dissolution results of test product(s) to the in vivo results.

Furthermore, such “successful” IVIVC outcomes, which are rare, are hardly used in practice to evaluate the quality of drug products. The procedures which are used for the evaluation of the quality of products (such as pharmacopeial tests) are generally not based on these IVIVC evaluations. This obviously adds to the frustrations as to why IVIVC studies are to be conducted when it may not be useful in assessing the quality of the product.

The question would then be, what is the intended purpose of the IVIVC practice? The intended purpose of the practice of IVIVC is not to develop (co)- relationship but to predict, more accurately estimate, a potential in vivo outcome. The in vivo outcome which is often used in this regard is drug concentration-time (C-t) profiles obtained from the BA/BE studies. Therefore, the objective of any dissolution testing must be to estimate and evaluate the C-t profiles. A detailed discussion on the procedural detail about developing such C-t profiles and their evaluations are beyond the scope of this article. Readers are referred to the literature on this subject, where necessary concepts and methodologies, in this regards, are provided in detail [9, 10].

»


« DIssolutIon

Choice of Experimental ConditionsAs dissolution test are conducted to evaluate potential drug release in vivo i.e., in the GI tract, choice of experimental conditions are, therefore, dictated by the physiological environment. Basically there are three variants which are usually considered in this regard: (1) temperature, which is 37 º C reflecting body temperature; (2) GI tract fluid which is reflected by water or aqueous solutions (buffers) having pH in the range of 5 to 7. If a drug is not expected to dissolve in water or buffers then a small amount of solubilizing agent may be added to enhance the solubility in the aqueous phase; (3) a mixing mechanism which is achieved by using a stirrer at a slow rotation speed. In short, water alone as a dissolution medium, or with small amount of solubilizing agent if the drug is of low aqueous solubility, maintained at 37 ºC with a stirrer at low rotation speed of 25 rpm may be used for testing of the majority of drug products [11]. It is to be noted that experimental conditions are derived from the physiological environment which remains the same from product to product thus these have to be product independent. However, a quick review of the literature shows that most experimental conditions, except temperature, are product dependent. Conducting dissolution studies using product dependent experimental conditions clearly negate the basic requirement of the testing. This creates a serious concern about the relevancy and credibility of current practices of dissolution testing, thus results obtained from dissolution testing would be of questionable merit.

At present there are two sets of variants in selecting experimental conditions for dissolution testing; media and apparatuses or stirrers. Commonly dissolution results are dependent on these two variants. In most cases, two types of apparatuses are used i.e., paddle and basket. These two types of apparatuses are similar in make and operation, expect for the stirring rods (or spindles). It is very well established, based on reports published in the literature, that these apparatuses are inherently flawed for dissolution testing because of poor hydrodynamics (mixing/stirring) within the vessels [2-4]. This flawed hydrodynamics results in serious deficiencies; such that the stirring provides limited product/medium interaction as well as creates unstirred and stagnant pockets. The physiological relevance of these apparatuses would thus be questionable as the intestinal environment provides thorough mixing and no stagnant pockets. Secondly, again based on the poor hydrodynamic characteristics, it has clearly been demonstrated that these apparatuses provide highly variable and unpredictable dissolution results unrelated to a product’s characteristics. Therefore, results obtained using these apparatuses will always be suspect and of limited use. There have been numerous attempts and suggestions for improving the behaviors of the apparatus by tightening specifications [12], but with little success as the issue does not appear to be with the specifications (tight or relaxed) but the apparatuses themselves.

Furthermore, as the cause of the problems is poor hydrodynamics within vessel using paddles and baskets, then, it may not be possible

www.sotax.com [email protected]@sotax.com

SOTAX Corporation 68A Elm Street Hopkinton MA 01748 USA Toll Free 1 888 SOTAXUS E-mail [email protected] Support [email protected] Tech Support Hotline 1 508 544 4040

Fully Automated Dissolution System

What SOTAX Automation customers are saying?

“I was asked to perform 80 method development lots including multiple rpms and buffers. Without SOTAX automation it would have taken me weeks to perform. It took me 4 days including data reporting”

“We have estimated a cost savings of over $250,000/year for our 3 highest volume products in our QC lab”

“The method included a time consuming full media change that took all day including analysis. With SOTAX automation, we reduced our testing time by 60%”

“We have noticed a signifi cant reduction in lab errors and investigations due to our SOTAX automation system”

“SOTAX has provided us excellent service and technical recommendations and solutions”

Are you ready to start saving? Contact your local SOTAX offi ce to schedule your demonstration.

»


DIssolutIon »

to make an appropriate choice of a dissolution medium using these apparatuses. The dissolution results obtained thus will always include high variability and unpredictability of the apparatuses. Unfortunately, instead of focusing on the issues of the apparatuses, there has been a tradition of supporting the use of paddle and basket apparatuses with weak rationales. The continued use of the paddle and basket apparatuses appears to be the major impediment of addressing the problems in developing appropriate dissolution tests [13].

looking to the FutureThe obvious question would be as to how these issues may be addressed. Obviously first and foremost is the need for recognition of the fact that unfortunately the recommended apparatuses (paddle and basket) are not appropriate for their desired purpose. There is strong experimental evidence in the literature regarding the deficiencies [5] as well as suggested solutions to address these [14]. However, there appears to be a lag in recognizing these developments. It is hoped that these new developments will provide impetus to re-evaluate the future use of paddle and basket apparatuses.

On the other hand, to accommodate the continued use of these apparatuses, at present, it has become a common practice to select arbitrary experimental conditions such as apparatus, rpm, dissolution medium etc. to achieve preconceived or expected dissolution characteristics of a product. The current practices of dissolution testing are therefore, in fact exercises of selecting/defining experimental conditions to obtain expected dissolution behavior rather than determining dissolution characteristics of the products. Hence, it would be safe to conclude that with the current recommended practices of dissolution testing one never determines the drug release (dissolution) characteristics of product.

In resolving the issue, it appears that there is a need for clearly defining and agreeing to the role of dissolution testing (evaluation of in vivo drug release) with an objective endpoint (developing C-t profiles). Such an objective and end point will facilitate the development/use of appropriate apparatuses and associated experimental conditions. One of the possible ways of achieving such an objective is through the availability of a reference product with known in vivo drug release characteristics. Such a reference product should be used in establishing the appropriateness of apparatuses and related experimental conditions. The use of such validated apparatuses and experimental conditions should be extended for other products. It is ironic that the drug dissolution community has been working for the past 3/4 decades and is expected to continue to work without a reference product. It is highly unlikely, if not impossible, that one will be able to get useful results from a technique/apparatus which has not be validated for its claimed propose. It is a critical deficiency which requires urgent attention.

In the absence of such a reference product, as well as for generating data towards developing a reference product, one may establish appropriateness of an apparatus and associated experimental conditions based on relative dissolution testing. The relative dissolution

testing may be described as determining drug dissolution (release) characteristics of two products of the same drug (active ingredient) but having two different known drug release characteristics in vivo such as IR and ER products. The dissolution test conditions should reflect a physiological environment and must be such that while providing different release patterns, fast for IR and slow for ER product, provide complete dissolution to occur within the suggested dosing interval for the drug products. Once such a set of experimental conditions is established, this may be considered as reflecting/simulating in vivo environment and then be used for other test products. It is to be noted that using experimental conditions which are not observed in vivo, such as de-aeration of dissolution medium, use of sinkers etc., be avoided as these may invalidate the testing.

In conclusion, it may be argued that most of the deficiencies/problems of current practices of dissolution may be related to poor hydrodynamics within the paddle and basket apparatuses which also lack relevance to physiological environment. The dissolution testing may significantly be improved if its role may clearly and objectively be established that the tests are to be conducted only to reflect in vivo dissolution characteristics of a product. This clarity of objective will provide an improved basis for selecting appropriate apparatuses and associated experimental conditions. In addition, such an objective will also reduce significant work load by eliminating requirements of repeated IVIVC developments and other physiologically non-relevant testing.

Disclaimer: Views expressed here are for scientific discussion purposes only and may not be reflective of opinions and policies of my employer (Health Canada).

references1. USP General Chapter on Dissolution <711>. United States

Pharmacopeia and National Formulary; United States Pharmacopeial Convention, Inc.: Rockville, MD, 2008. p.267-274.

2. Baxter JL, Kukura J, Muzzio FJ. Hydrodynamics-induced variability in the USP apparatus II dissolution test. Int J Pharm 2005;292:17-28.

3. Healy AM, McCarthy LG, Gallagher KM et al. Sensitivity of dissolution rate to location in the paddle dissolution apparatus. J Pharm Pharmacol 2002;54:441-4.

4. Qureshi SA, McGilveray IJ. Typical variability in drug dissolution testing: Study with USP and FDA calibrator tablets and a marketed drug (glibenclamide) product. Eur J Pharm Sci 1999;7:249-258.

5. Qureshi SA. A new crescent-shaped spindle for drug dissolution testing - but why a new spindle? Dissolution Technol 2004;11:13-18.

6. D’Souza1, S.S., Lozano, R., Mayock, S., and Gray, V. AAPS Workshop on the Role of Dissolution in QbD and Drug Product Life Cycle: A Commentary. Dissolution Technol. 2010, 17 (4), 41-45.

7. Tong, C., Lozano, R., Mao, Y., Mirza, T., Löbenberg, R., Nickerson, B., Gray, V., and Wang, Q. The Value of In Vitro Dissolution in Drug Development A Position Paper from the AAPS In Vitro Release and Dissolution Focus Group. Pharmaceutical Technology 33 (4), 52-64 (2009).

8. US FDA Guidance for Industry (1997): Extended Release Oral Dosage Forms: Development, Evaluation, and Application of In Vitro/In Vivo Correlations. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM070239.pdf (Accessed December 29, 2010).

»


« DIssolutIon

9. Qureshi, S.A. In vitro-in vivo correlation (IVIVC) and determining drug concentrations in blood from dissolution testing – A simple and practical approach. The Open Drug Delivery Journal, 2010, 4, 38-47. (Link). (Accessed December 30, 2010).

10. Qureshi, S.A. Determining blood concentration-time (C-t) profiles from in vitro dissolution results and product evaluation – carbamazepine. http://www.drug-dissolution-testing.com/?p=601 (Accessed December 30, 2010).

11. Qureshi SA. Drug dissolution testing: Selecting a dissolution medium for solid oral products. Am Pharm Rev 2009;12:18-23.

12. Gray, V.A. Identifying Sources of Error and Variability in Dissolution Calibration and Sample Testing. Am. Pharmaceutical Reviews. 5:2 (2002) 8-13.

13. Gray, V., Kelly, G., Xia, M., Butler, C., Thomas, S. and Mayock. S. The Science of USP 1 and 2 Dissolution: Present Challenges and Future Relevance. Pharmaceutical Research, 26:6, 2009, 1298-1302.

14. Qureshi SA. A Crescent-shaped Spindle for Improved Dissolution Testing. Pharmeuropa Bio & Scientific Notes, 1:2009: 55-66.

Author BiographyDr. Qureshi is a senior research scientist in the Therapeutic Products Directorate, Health Products and Food Branch, Ottawa, Canada. His main area of research involves the assessment of drug release characteristics, both in vitro and in vivo, of oral and dermal products. He has published more than 40 papers, including a chapter in “Encyclopedia of Pharmaceutical Technology” as well as made numerous national and international presentations in the areas of drug dissolution testing, analytical chemistry, pharmacokinetics, bioavailability and bioequivalence. Dr. Qureshi, moderates and is a frequent contributor to a blog on the subject (www.drug-dissolution-testing.com).

Reach your target.If you need authoritative, unbiased literature to enhance your product or service, what better way than to order reprints of one of the articles in this or any issue of American Pharmaceutical Review. Professionally presented with an advertisement or simply your contact details, reprints are a great way to showcase your company.

For more information about purchasing reprints, contact us at 317-816-8787 or at [email protected]

»


mICroBIology »

Kalavati Suvarna, Ph.D, Anastasia Lolas, M.S. Patricia Hughes, Ph.D. & Richard L Friedman, M.S.Biotechnology Manufacturing Team, Division of Manufacturing and Product Quality,Office of Compliance, Center for Drug Evaluation and Research, Food and Drug AdministrationEmail: [email protected].

Case Studies of Microbial Contamination in Biologic Product Manufacturing

AbstractThe manufacture of biologic products is a complex process and requires the use of living cells. These processes and products are prone to contamination by adventitious agents such as bacteria, fungi and viruses. Microbial contaminations have a huge impact on biologic product manufacture as they introduce product variability and can cause loss of potency due to degradation or modification of product by microbial enzymes, changes in impurity profiles, and an increase in the levels of bacterial endotoxins. In addition, the investigations of microbial contaminations can result in lengthy shutdown periods and delays in manufacturing operations that in turn, may sometimes result in shortages of essential drug products. Strict microbial production controls are essential to ensure the manufacture of a drug product with consistent quality. This article discusses elements of a microbial control strategy, recent cases of microbial contamination in specified biologic products, the need to perform risk assessments on a periodic basis, and additional areas of improvement in the management of risks.

IntroductionBiologic products are manufactured using living cells such as bacteria, yeast, and mammalian cells. These include specified biologics such as monoclonal antibodies and therapeutic recombinant DNA-derived products licensed under Section 351 of the Public Health Service Act [1] and currently regulated by the Center of Drug Evaluation and Research (CDER). These biological products are also regulated as drugs under the Federal Food, Drug, and Cosmetic Act [2]. The upstream process in the manufacture of monoclonal antibodies and therapeutic recombinant proteins typically involves cell expansion, cell culture, and recovery steps. The downstream process involves multiple purification steps. The purified protein is ultrafiltered/diafiltered with formulation buffer to provide a formulated bulk drug substance. The formulated bulk drug substance is sterile-filtered and filled to provide a final drug product. Because of the consequences of microbial contamination on product safety and quality, there is continued interest in understanding the root causes of microbial contamination and controlling these risks in biologic product manufacture. This article discusses some of the bacterial contamination cases reported to the Agency or identified during pre-license/pre-approval inspections of biologic drug substance manufacturers in the past two years. The cases highlight areas for

Innovation is in Our Genes

PyroGene™ rFC AssayThe evolution of endotoxin detection.n Endotoxin specific, recombinant technology eliminates false-positive glucan reactions

n Predictable, reliable assay performance lot after lot

n Sustainable resource – no animal utilization

n Endpoint fluorescent assay, comparable to other quantitative LAL methods

n 510(K) submissions have been approved by the FDA, using PyroGene™ rFC Assay as a final release test

Discover for yourself. To evaluate the PyroGene™ Recombinant Factor C Assay, visit www.lonza.com/rfc.

Lonza Walkersville, Inc. 8830 Biggs Ford Road, Walkersville, MD 21793800 654 4452 301 898 7025PyroGene is a trademark of the Lonza Group or its affiliates. ©2010 Lonza Walkersville, Inc.

PyroGene_Ad_AmerPharma_rev_0111.indd 1 1/18/11 10:45 AM

»


mICroBIology »

improvement in risk management and the need for developing a robust microbial control strategy for biologic products.

sources of microbial ContaminationMicroorganisms are ubiquitous in nature. Microorganisms can adapt and survive under a variety of conditions and can pose a significant risk to biologic products. An understanding of the microbial entry points and implementation of measures to prevent microbial contamination is critical for manufacture of safe, pure and potent biologic products. As shown in Figure 1, microorganisms can gain entry into a production process stream from several sources: the facility, equipment, process operations, raw materials, column resins, filter membranes, water, process gases, and personnel. All sources of microbial contamination should be considered when developing a microbial control strategy and performing an investigation for a microbial contamination deviation.

regulation and guidanceThe minimum current good manufacturing practice (CGMP) requirements for preparation of finished human drug products are described in 21CFR§211 [3]. These include the use of suitable protective apparel (21CFR§211.28), appropriate facility design and placement of equipment (21CFR§211.42), equipment cleaning, sterilization, and maintenance (21CFR§211.67), and production and process controls (21CFR§211.100). All these preventive measures and precautions are implemented to protect product and prevent contamination. Therapeutic recombinant products and monoclonal antibodies are also subject to applicable regulation in 21CFR parts 600-610 [4]. The guidance on CGMP for active pharmaceutical ingredients, Q7A, provides general CGMP guidance for biologic drug substance manufactured by cell culture or fermentation under section XVIII [5]. Additional guidance documents cover prevention or control of adventitious agents in cell-derived biologic products and

address the quality concerns originating from cell substrates used for manufacture of these products. These documents include (a) Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use published in February 1997, (b) Guidance on Viral Safety Evaluation of Biotechnology Products Derived From Cell Lines of Human or Animal Origin (Q5A), (c) Guidance on Quality of Biotechnological/Biological Products: Derivation and Characterization of Cell Substrates Used for Production of Biotechnological/Biological Products (Q5D), and (d) Guidance on Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products (Q6B) [6,7,8,9]. The Q6B guidance states that contaminants should be strictly avoided and/or suitably controlled with appropriate in-process acceptance criteria or action limits for the drug substance or drug product to meet specifications. The 1994 FDA Guidance for Industry on the Submission Documentation for Sterilization Process Validation in Applications for Human and Veterinary Drug Products provides guidance on sterilization process validation for final drug product [10]. The 2004 FDA Guidance for Industry on “Sterile Drug Products Produced by Aseptic Processing – Current Good Manufacturing Practice” provides guidance on personnel qualifications, clean room design, process design, and aseptic processing of final drug products [11]. These regulations and guidance documents provide the backbone for the development of an appropriate microbial contamination control strategy.

Elements of a microbial Control strategyA microbial control strategy should be developed once a comprehensive risk assessment has been performed for all possible microbial entry points into the manufacturing process. This requires a good understanding of the manufacturing process and product attributes. In general, the design of the facilities should allow for proper operations and prevention of contamination. The flow of personnel, material and waste should be from clean to dirty areas and critical upstream open operations liable to microbial contamination should be performed in designated biosafety hoods or areas with ISO 5 classification. Depending on the risks to the process, areas should be appropriately segregated. Segregation of pre-viral and post-viral clearance steps in processes using mammalian host cells is important to prevent cross-contamination of process intermediates and the facility. Segregation of areas, appropriate changeover procedures, and other procedural controls should be in place to prevent cross-contamination in a multi-product facility. Environmental monitoring of manufacturing areas should be performed routinely at appropriate intervals. Process gases and water should be tested and monitored to ensure adequate microbial control. The design of equipment (single-use disposable versus multi-use), validated cleaning and sterilization processes along with a comprehensive preventative maintenance plan are critical components of the microbial control strategy. Microbial control for the lifetime use of membranes and resins should be demonstrated. In addition, it is critical to identify and establish processing steps that decrease bioburden and bacterial endotoxin levels as the process intermediates are processed through sequential purification steps. Bioburden reducing filters should be used at

Figure 1: Sources of microbial contamination.

124 Bernard E. Saint Jean Drive, East Falmouth, MA 02536 USAtel: 888.395.2221 • fax: 508.540.8680 • [email protected] • www.acciusa.com

Run Both Chromogenic AND Turbidimetr ic Assays in

The Same K inetic Tube Reader

F L E X I B I L I T Y

»


mICroBIology »

critical steps in the process. This is critical for buffer solutions and in process intermediates conducive to microbial growth. Minimizing the number of open operations reduces the risk to product from external (personnel and environmental) microbial contamination sources. Biologic products are usually rich in carbon sources that favor microbial growth. Hold conditions (time, temperature) for a process should be validated to control and prevent potential microbial growth. Bioburden and endotoxin alert and action limits should be set for process steps based on process capability. Raw materials should be screened for microbial quality and should be handled and stored in a manner to prevent contamination and cross-contamination. Personnel are important contributors to microbial contaminations. Appropriate gowning should be implemented to prevent contamination. All personnel performing open operations should be trained adequately and evaluated periodically in such operations.

Case studiesIn the last two years, several contamination events were reported to the Agency. They included viral or bacterial contamination of upstream cell culture or fermentation processes. Viral contamination events were extensively covered in the recent 2010 PDA/FDA Adventitious Viruses in Biologics: Detection and Mitigation Strategies Workshop. Only

bacterial contaminations are discussed in this article. One case involved contamination of a fermentor used in the manufacture of a protein product secreted by a bacterial host. The contaminant was identified as Bacillus cereus (a Gram positive spore forming rod). A second case involved the contamination of a fermentor used in the manufacture of a recombinant protein by Paenibacillus curdlanolyticus (a Gram variable spore-forming rod). A systematic approach was used during the investigations to identify the root cause of the contamination and included several media simulations to aid in identifying the point of entry into the fermentor. In addition, the investigations involved the manufacture of engineering batches. After a lengthy investigation in both case studies, problems with the sampling devices, addition valves, incorrectly fitted components, missing O-rings, incorrect installation and deformation of an air filter after sterilization, and/or inadequate slope of a condensate line were identified. Immediate corrective actions included the replacement of valve diaphragms in fermentor addition ports, replacement of a membrane valve in the sampling device, and replacement of O-rings on the measuring probes. Enhancements were also made to the sterilization processes of fermentor and associated transfer lines. A preventative maintenance plan was developed for all fermentor valves. All valves were tagged using a detailed checklist to ensure correct installation. All SOPs were updated and employees were trained on the revised versions. The investigations and corrective actions addressed all possible causes of contamination as an unequivocal root cause could not be assigned.

In most cases, it is very difficult to identify a definitive assignable cause. It is highly recommended that a systematic approach be followed to determine the root cause. Media simulations help in demonstrating that sterility of the fermentor is not compromised. Recent microbial contamination events at several manufacturing facilities point to breaches in the sterile boundary caused by damaged vent filters, damaged O-rings, diaphragms, and elastomers, and improperly sloped condensate lines.

When bacterial hosts are used, microscopic examinations of the fermentation culture for contamination is difficult. A culture purity test should be perfomed using appropriate media and culture conditions. It is crucial to have a comprehensive preventative maintenance plan for fermentor and tank agitators, probes, gaskets, O-rings, valves, and filters. The design of piping and valves should prevent steam condensate from collecting and leading to contamination by back-flow. After periods of shutdown or maintenance, it is important to perform media simulations on sterile equipment that has remained idle for a period of time. Procedural details on assembly and set-up of fermentors/bioreactors should be clear and very detailed. Training in this area can reduce inadvertent

AccuPRO-ID™ is the ideal choice for routine environmental monitoring andprimary screening of bacterial organisms. Backed by our validated library,

AccuPRO-ID™ outperforms the VITEK® 2 Compact by 2X in accuracy.

• Higher accuracy at lower cost to customers currently using any phenotypic method

• Utilizes our validated, cGMP libraries, based on testing hundreds of thousands of industry-relevant organisms

• Samples that do not produce a result using MALDI-TOF will automatically be sequenced for an ID at no additional charge

Try us out for free! Simply submit up to 5 bacterial samples and write the Redemption Code: “APR-AD” in the “Comments” column on the Identification Request

Form. To access the form, please visit us at http://www.accugenix.com/info_ref.phpand download “AccuPRO-ID IRF” or call +1.302.292.8888.

Image: Copyright Dennis Kunkel Microscopy, Inc,.

n = 60 unknownenvironmental isolates (46 different species)

ASM 2010 Poster I-2664

Discover the Power

IntroducingAccuPRO-ID™

AccuPRO-ID™ vs. VITEK® 2 Compact

»


« mICroBIology

leaks and contamination of the systems. Continuous assessments of change control, work orders, and other process improvements should be conducted to ensure that the microbial control strategy is not impacted. Of note in both cases, the contaminating microorganism was a facultative anaerobic Gram positive spore-forming rod. Risk mitigation strategies based on microbial environmental flora should be considered. The areas for improvement identified in the case studies were in preventative maintenance plans for all fermentor valves including valves on sampling devices and in the documentation for correct assembly of components.

Two cases of microbial contamination of the downstream process were identified during pre-approval/pre-license inspections of drug substance manufacturing facilities. Bioburden deviations were observed in several batches at the ultra-filtration/diafiltration (UF/DF) step. The contaminants identified were Sphingomonas species, Stenotrophomonas maltophilia, Ralstonia pickettii, and Staphylococcus species suggesting probable water and human sources of contamination. Presence of repeated high bioburden counts in several batches suggested development of biofilm and inadequate contamination control procedures for the UF/DF steps. After extensive investigations, several corrective actions were implemented in terms of cleaning, storage and re-use of UF/DF systems, sterilization/sanitization of buffer tanks, assessment of the water for injection (WFI) system and transfer lines, introduction of in-process bioburden reducing filters (in cases where there were no filters before the UF/DF steps), validation of hold times and storage conditions of process intermediates and revisions to bioburden limits based on process capability. Demonstration of microbial control over the lifetime use of membranes and validation of in-process hold times are essential for ensuring the consistent quality of biologic products. All WFI piping locations with stagnant water should be assessed and eliminated. Microbial trend reports for water systems should be reviewed regularly.

The investigations of microbial contaminations are challenging due to the ubiquitous nature of the microorganisms, multiple points of microbial entry, growth promoting properties of biological process streams, limitations of sampling and detection methods, and the time and resources involved in performing complex investigations. All microbial entry points should be systematically evaluated. For fermentor contaminations, seed fermentors and associated additions and transfer lines should be included

in the investigations. A hazard analysis and critical control point assessment for bioburden control throughout the manufacturing process is useful for the design of a microbial control strategy and the performance of a systematic investigation. In addition, failure data should be tracked to gain a better understanding of root causes. The information should be used to continuously evaluate risks and implement process and/or equipment improvements to mitigate and prevent microbial contaminations.

Accelerate and automate your microbial testing.

The Growth DirectTM SystemRapid Automated Compendial TestNon-Destructive

Replaces human eye with digital imaging to deliver rapid microbial enumeration:

Automated incubation, enumeration, reporting •& LIMS data transfer saves labor & improves compliance

Applications & throughput cover your routine •QC testing needs

Streamlined validation – compatible with •USP methods and regulatory guidelines

Non-destructive test allows faster investigations•

Rapid Micro Biosystems, Inc.Bedford, MA 01730

Phone: 781-271-1444www.rapidmicrobio.com

Save time. Save money. Increase productivity.

»


mICroBIology »

ConclusionsMicrobial contamination is a risk to biologic product quality and safety. The cost of inadequate microbial control in biologic product manufacture is enormous as facilities or bioreactor production trains may have to be shut down for lengthy periods of time in order to conduct investigations and identify the root cause to prevent reoccurrence. The recent cases of bacterial contamination of biologic products suggest that preventative maintenance plans for fermentor and associated valves, types of materials used for diaphragms and O-rings, and understanding of microbial control at certain process steps need further attention. Contamination control requires an understanding of the microbial entry points and risks to the process as well as the microbial growth potential of the product, media and buffer solutions. Microbial contamination control requires appropriate design of facility and equipment, validated cleaning and sterilization cycles for equipment, detailed and robust preventative maintenance plans for equipment, measures to reduce bioburden and bacterial endotoxins at appropriate steps in the process, and routine monitoring of these process steps for bioburden and endotoxin with defined alert and action limits. A contamination remediation plan should be established. Such a plan is beneficial for meeting CGMP and has the advantage of reducing facility downtime. Investigations should be comprehensive and include assessment of all microbial entry points. Corrective actions should address all possible identified causes in the absence of a known assignable root cause. The information gathered during these investigations should feed into the overall risk management plan. The quality risk management plan should be integrated into the quality system and allow for continuous improvement.

references1. Public Health Service Act, Biological Products; as amended

2. Federal Food Drug and Cosmetic Act; as amended.

3. FDA, “Current Good Manufacturing Practices for Finished Pharmaceuticals,” 21 CFR part 211.

4. FDA, “Biologics,” 21 CFR parts 600-610.

5. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry: Q7A Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients. Rockville, MD; 2001.

6. U.S. Department of Health and Human Services, Food and Drug Administration. Centre for Biologics Evaluation and Research. Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use. February 1997.

7. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry: Q5A Viral Safety Evaluation of Biotechnology Products Derived From Cell Lines of Human or Animal Origin. Rockville, MD; 1998.

8. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry: Q5D Guidance on Quality of Biotechnological/Biological Products: Derivation and Characterization of Cell Substrates Used for Production of Biotechnological/Biological Products. Rockville, MD; 1998. Federal Register Vol. 63, No. 182, 1998.

9. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry: Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products, FDA, 1999.

10. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry for the Submission Documentation for Sterilization Process Validation in Applications for Human and Veterinary Drug Products. Rockville, MD; 1994.

11. U.S. Department of Health and Human Services, Food and Drug Administration. Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing – Current Good Manufacturing Practice. Rockville, MD; 2004.

BiographyKalavati Suvarna, Ph.D. is a Microbiologist with the Biotech Manufacturing Team in the Division of Manufacturing and Product Quality in the Office of Compliance, CDER, FDA. She has over nine years of experience as a microbiology reviewer at the FDA. Kalavati holds a Ph.D. in Biological Sciences from Northern Illinois University. Prior to joining the Agency, she worked in an academic and pharmaceutical setting.

Anastasia G. Lolas is a Microbiologist with the Biotech Manufacturing Team in the Division of Manufacturing and Product Quality in the Office of Compliance, CDER, FDA. She has over 5 years of experience as a microbiology reviewer of drug applications at the FDA. Anastasia holds a B.S. in Biology from Virginia Polytechnic Institute and State University and a M.S. in Food Science from the University of Illinois at Urbana-Champaign.

Patricia F. Hughes, Ph.D. is the Team Leader in the Biotech Manufacturing Team in the Division of Manufacturing and Product Quality in the Office of Compliance in CDER, FDA. She has over twenty years experience in the Pharmaceutical/Biotech industry in fermentation & cell culture process development and manufacturing. In addition, she has over twelve years of experience as a microbiology reviewer at the FDA, in CDER and CBER. Patricia holds a Ph.D. in Microbiology from Georgetown University.

Richard Friedman is the Director of the Division of Manufacturing & Product Quality in the Center for Drug Evaluation and Research (CDER), Office of Compliance. In this position, he directs the interpretation and development of CGMP policy, review of inspectional recommendations and determination of manufacturing site acceptability. He has been employed by FDA since 1990, including prior positions as New Jersey District Drug Specialist, CDER Senior Compliance Officer and Team Leader of Guidance and Policy. Mr. Friedman has authored several publications on topics including sterile drugs and quality management systems, and was awarded The George M. Sykes Award by the Parenteral Society for outstanding journal paper for the year 2005. Mr. Friedman is also an adjunct faculty member of Temple University School of Pharmacy in their QA/RA graduate program. Prior to joining FDA, Mr. Friedman worked in the toxicology research division of an innovator pharmaceutical company. Mr. Friedman received his B.S. in Biology with honors from Montclair State University in 1989 and his M.S. in Microbiology from Georgetown University School of Medicine in May, 2001.

Do you live an InterContinental life?

Whether you’re in Baltimore for business or pleasure, a stay at the four-star InterContinental Harbor Court Baltimore Hotel is guaranteed to leave you with an unparalleled experience of the city. With our award-winning restaurants, impeccably appointed rooms and enticing amenities, the Harbor Court Baltimore reveals “the beauty of Baltimore” to guests at every turn. And with our central location in the heart of the historic Inner Harbor, within walking distance to the downtown business district and countless popular attractions, there’s no easier way to access everything you require to make your visit a success. Get the most from your Baltimore experience, the InterContinental way.

A STEP A BOV E THE R EST, IN THE HE A RT OF BA LTIMOR E’S INNER H A R BOR.

Call 1.800.424.6835 or visitwww.intercontinental.com/baltimore©2007 InterContinental Hotels Group. All rights reserved.

.

»


sInglE-usE »

Disposable manufacturing platforms are becoming increasingly popular for therapeutic monoclonal antibody (MAb) manufacture, whether as fully disposable platforms or for incorporating single use solutions in to largely fixed (i.e., stainless steel-based, re-usable) manufacturing platforms. While large pharmaceutical companies have largely adopted a platform approach to development and commercialisation of therapeutic monoclonal antibodies, or employ single use solutions for certain unit operations (e.g., seed train expansion), disposable platforms may be more common among small to medium sized, or more recently established, companies. This may have implications for partnerships between small-to-medium and large pharma companies seeking to co-develop and commercialise biopharmaceuticals.

We describe here the transfer, manufacture of clinical trial drug substance lots, and late stage development in advance of process validation, of a fully disposable MAb manufacturing process in an existing small molecule API clean room facility. Four MAb drug substance lots were manufactured within 14 months of facility selection decision. The overall transfer process, and specific challenges arising from implementing disposable-based biopharmaceutical manufacturing in an existing small molecule API facility are discussed. This approach may become more relevant as companies assess existing facilities in the context of a changing business environment (moving in to biopharmaceutical manufacturing, strategic responsiveness to partnering opportunities). The case described here represents an alternate route to biopharmaceutical manufacturing, creating options for leveraging existing facilities, as opposed to constructing dedicated biopharmaceutical manufacturing space.

The MAb molecule discussed here is a therapeutic IgG1, manufactured by a GS-CHO-based (fed-batch), two column process. The molecule was co-developed between Eli Lilly & Co. and MacroGenics, Inc. An overview of the process is shown in Figure 1. The process is based largely on single use components, with the affinity capture chromatography column and purification skids being re-used (i.e., cleaned between uses).

As shown in Figure 2, Eli Lilly began co-developing the MAb for commercialisation in 2008. At this time, MacroGenics had already generated material for Phase III supply, and additional Phase III material was to be generated in Lilly’s Kinsale facility, which would also be the site for process validation, launch, and commercial supply. Lilly Kinsale has a 30+ year history in small molecule API market supply (post-launch) for both parenterals and non-parenterals and was in

Brian Mullan, Ph.D., Kristi Huntington, Aidan Collins, and Marie Murphy, Ph.D. Eli Lilly & Co.email: [email protected]

Transfer, Implementation and Late Stage Development of an End-To-End Single-Use Process for Monoclonal Antibody Manufacture

w: www.pall.com/economicse: [email protected]

For those who appreciate a high degree of

selectivity

Defining Process Economics© 2011 Pall Corporation. Pall, , HyperCel, and Mustang are trademarks ofPall Corporation. ® indicates a trademark registered in the USA. GN10.3516

Unique chromatographyselectivities deliver economy and performance at every scale

Pall’s chromatography chemistries featuredifferentiated selectivities for ion exchange and mixed-mode chromatography that help achieve protein purification goals and decrease process costs.

u Increase productivity: Q and S HyperCel™

sorbents provide high dynamic binding capacity at 2 to 5 times the flow rates ofconventional ion exchangers.

u Reduce process steps: HyperCel mixed-mode sorbents can eliminate unit operation steps from your process.

u Decrease salt concentration: MEP, HEA and PPA HyperCel sorbents are an alternative to HIC that reduces the amount of salt required for protein capture.

u Decrease buffer usage: Mustang® ion exchange membrane capsules significantly reduce buffer consumption during contaminant removal.

u Simplify scale up: Flexible designs allowconsistent performance from lab to process scale and reduce revalidation requirements.

Visit Pall online to get the complete story.

10-3516_Chrom-AmericanPharmRev-Fullpg_AD:layout 17/01/2011 12:34 Page 1

»


sInglE-usE »

the process of developing launch and commercial supply capability for biopharmaceuticals at this time (2007; construction of a re-usable platform-based facility in Kinsale). The agreement and project timing with MacroGenics was in advance of scheduled availability of this new facility (2010). Additionally, this opportunity enabled Lilly Kinsale to begin to build capabilities in the areas required to support biopharmaceutical manufacturing, using a smaller scale and flexible platform, in advance of its larger scale platform facility coming online in 2010.

Figure 1. Process flow for MAb manufacturing process

Figure 2. Overall timeline for process transfer, facility readiness and schedule of Phase III campaigns leading to process validation in Q4 2010. The red diamond indicates timing of facility selection

decision. Activities at Lilly’s Kinsale facility are shown in blue boxes, activities at Lilly Bioprocess R&D, Indianapolis, are shown

in orange boxes. Drug substance technical program (green boxes) was jointly executed between Kinsale and BR&D. DS,

drug substance; BR&D, Bioprocess R&D; CT, clinical trial; ENG, Engineering batches; PS, primary stability; PV, process validation;

SM, small molecule.

GE HealthcareLife Sciences

Part of the ReadyToProcessTM platform, WAVE Bioreactor is a fast and efficient system for innoculum propa gation and cell culture. Scale-up is simple as no passaging is needed – just add fresh culture media to the Cellbag™ bioreactor. WAVE Bioreactor reduces time-consuming routines, increases flexibility, and ensures the integrity of your operations. ReadyToProcess, just plug in & you’re ready to go.

Find out more at www.gelifesciences.com/readytoprocess

Plug & play with WAVE Bioreactor™

and accelerate your seed train

Cellbag, ReadyToProcess, and WAVE Bioreactor are trademarks of GE Healthcare companies.© 2010 General Electric Compnay – All rights reserved. GE Healthcare Bio-Sciences AB, Björkgatan 30, 75184 Uppsala, Sweden.

GE10-10. First publised November 2010.

»


« sInglE-usE

At a process development level, the process as transferred was appropriate for Phase III supply, but required additional characterisation and development to be ready for process validation. This work was initiated in Eli Lilly’s Bioprocess R&D (BR&D) facility in 2008, with co-development a joint effort between Lilly’s BR&D and Kinsale development units.

Numerous considerations were assessed to transfer a large molecule manufacturing process in to a clean room facility previously used for a small molecule parenteral API (Table 1). Among these, key considerations to meet the overall timeline were supply chain and analytical testing. For the first (Engineering / Clinical Trial supply) Phase III campaign in Kinsale, the supply chain was managed by BR&D, Indianapolis, with raw materials and consumables being transferred to Kinsale from Indianapolis via inter-warehouse transfer (followed by receipt verification and GMP storage in Kinsale). All analytical testing for the MAb molecule (batch release, in process) was performed by MacroGenics, Lilly BR&D, or was outsourced (e.g., for adventitious agent testing). The exception to this was microbiological testing (bioburden, endotoxin), for which methods were transferred to and/or qualified in Kinsale to support direct testing at the Kinsale site, and raw materials release testing, which was also performed at Kinsale.

At a facility level, the overall approach was to establish a GMP system for campaign-based use of an existing small molecule API parenteral facility on the Lilly Kinsale site. This facility consisted of in-built equipment (e.g., glove boxes) in numerous, large (ca. 40m2 per clean

room) ISO 7 or 8 classified clean areas. During small molecule API manufacture intermediates are housed and moved in closed transport containers between unit operations. The facility contained a parts washer and an autoclave, and dedicated processing areas for different areas of process support (e.g., non-clean and clean parts areas), and for different unit operations of the API manufacturing process.

For biopharmaceutical manufacturing, areas in the facility were assigned as follows:

Small, simple, precise pumps

The compact 120peristaltic pump forbiopharm, science andresearch saves benchspace and improves yield.Exceptional 2000:1 speedrange offers precise flowsto 190ml/min.

NEW

Watson-Marlow Flexicon Tubing wmpg.com800-282-8823

BioPharmaceutical Division

See us

at Interphex

booth #2807

120-WM-APR7x4.875_Layout 1 2/9/11 9:15 AM Page 1

Table 1. Facility fit and other considerations for transfer of a MAb manufacturing process in to a small molecule parenteral API

manufacturing facility. Items are shown in alphabetical order. DCS, Distributed control system; HVAC, Heating ventilation and air

conditioning; SME, subject matter expert;• Alarm management (accessibility to existing system, ability to route alarms to supervisory

areas)

• Autoclave, parts washer availability

• Cold storage (2-8oC, -20oc, -80oC)

• DCS points / accessibility to plant historian system

• Operations readiness and training

• Personnel and equipment flows

• Potential cross-contamination with existing products (multi-product management)

• Quality systems

• Resources, capabilities (SMEs, technical team, Analytical [in-house or out-sourced])

• Utilities (HVAC, number of power points, process gases)

• Viral boundary (pre-, post-)

• Warehousing space

• Waste management (including biohazardous waste) and disposal

»


sInglE-usE »

• Buffer preparation* (ISO 8)

• Cell culture media and feed preparation* (ISO 8)

• Vial thaw and seed train expansion (ISO 7 with ISO 5 biological safety cabinet added)

• Production Bioreactor and Primary Recovery (ISO 8)

• Initial purification: Affinity capture to Nanofiltration (ISO 8)

• Final purification: UFDF to final formulation and fill (ISO 8)

* weigh and dispense, dissolution, in process checks.

A pre-viral/post-viral boundary, controlled by GMPs, was designated between the last two areas.

As the existing facility contained adequate open spaces, supporting manufacture of the small molecule parenteral API largely through the use of fixed equipment, the change-overs to support biopharmaceutical manufacturing (Figure 2) were based largely on a “wheel-in, wheel-out” approach, with some facility modifications, as shown in Figure 3.

specific Challenges: supply Chain and Analytical testingThe small molecule process supported by the Kinsale clean room facility had about five raw materials and 15 consumables. The MAb manufacturing process had about 20 raw materials and 200 consumables, ranging in size from disposable bioreactor bags (fully packaged) down to pipette tips. Logistic and supplier quality aspects of supply chain were supported by Lilly Indianapolis (with inter-warehouse transfers to Kinsale) as establishment of a supply chain de novo for Lilly Kinsale’s first biopharmaceutical would not have been possible in the agreed project timeline. Even with this support, physical storage and management of this amount of raw materials and consumables was challenging, with the MAb process effectively taking over an entire warehouse, including a walk in 2 – 8oC area, and required installation of -20oC and -80oC storage.

The Lilly Kinsale site had a well established Analytical group supporting small molecule manufacturing, including microbiology testing and environmental monitoring (EM). Transfer and validation of the ca.15 methods supporting release testing of the MAb process would not have been possible in the agreed timeline as the majority of methods required new equipment (e.g., for CE-LIF, CEX, CE-SDS methods) and expertise. All testing, excepting EM and microbiological testing, and raw material release testing, was performed by MacroGenics, Lilly BR&D, or was out-sourced (for adventitious agent testing and bioassay).

the new math ofrapid detection

The Cell Culture Rapid Methods Program from Life Technologies lets you detect the three most prevalent threats to CHO cell culture manufacturing, typically in less than 4 hours, enabling near–real-time monitoring from start to finish.

1 sample prep + 1 instrument platform =detection of Mycoplasma, Vesivirus, and MMV

DNA | RNA | PROTEIN | CELL CULTURE | INSTRUMENTS

Go to www.appliedbiosystems.com/seq

FOR RESEARCH USE ONLY. NOT INTENDED FOR ANY ANIMAL OR HUMAN THERAPEUTIC OR DIAGNOSTIC USE. © 2011 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. CO17317 0210

CO17317 CCRM Ad 1/2page Resize for American Pharma.indd 1 2/10/11 5:05 PM


»« sInglE-usE

specific Challenges: Disposable ProcessesNo specific challenges were encountered during implementation and execution of unit operations where disposable technologies are more routinely applied, e.g., seed train expansion, primary recovery. Issues more so arose with the production bioreactor, some protein purification unit operations, and generic concerns related to use of disposable manufacturing systems.

At a generic level, sourcing and availability of standard tubing assemblies (extender pieces, pump tubing assemblies) aided process flow during manufacturing of the initial (non-GMP) engineering lots, enabling set-up of equipment trains to be optimised (process fit and connection harmonisation). Off-the-shelf tubing solutions are not widely available from vendors, and custom solutions from most vendors can take 4 – 16 weeks for delivery, by the time vendor set-up, assembly design, fabrication, γ-irradiation are complete. Additionally, key technical details for consumables can become buried in the large amount of items (in our case 200+) that need to be simultaneously set up and managed. An example in point is a key filter for protein purification, that at the original 1.2m2 configuration was supplied as γ-irradiated by the vendor, but at 0.6m2 configuration (as used at Lilly Kinsale) was not supplied as γ-irradiated. This aspect was not initially noticed during materials set-up. In this instance, an additional γ-irradiated 0.2/0.45um guard filter was placed in line (downstream) of the 0.6m2 filter to provide additional aseptic assurance.

For the production bioreactor, the process as transferred comprised a disposable cell bag from one vendor installed in to a bioreactor system from another. Initial interest in moving to in-line probes (for pH and temperature) was not supported by our experience with these probes (numerous bag ruptures), and a customised probe was qualified for temperature control, with pH being monitored off-line via a blood gas analyser.

For the purification unit operations, the process as transferred contained some manual mixing steps, which were all replaced by non-manual mixing systems (rocking platforms, bulk mixing systems). This necessitated mixing development and optimisation of control parameters in many instances.

Lastly, the platform was further developed and optimised by developing customised single use solutions for certain issues encountered during process transfer or which were required prior to process validation. These included:

• Design of a integral filter + bag solution (and sampling port assembly) for intermediate hold of clarified harvest;

• Design of a bulk mixing system for pH adjustment of a purification intermediate;

• Integration of individual components required for final fill and filtration of formulated drug substance in to a single γ-irradiated assembly.

single use systems

Contact us by calling

800-724-4158

As a part of Advanced Scienti cs’ Single Use Systems, the company offers a multitude of solutions for single use bags. With sizes ranging from 50 mL to 3,500 L, Advanced Scienti cs can produce single use bags of nearly any con guration. These bags are currently utilized throughout the industry and throughout the manufacturing process.

In addition to specializing in custom solutions, we offer a large variety of catalog products.

Advanced Scienti cs is an FDA registered, ISO 13485:2003 certi ed manufacturer.

ww

w.ad

vanced

scienti cs.co

m

Figure 3. Adaptation of an ISO 8 classified area in a facility used for small molecule parenteral API manufacturing. [Left Panel] Open space is shown, containing glycol loops. [Right panel]

200L disposable bioreactor systems in place in same area. A process gas (air, O2, CO2) manifold has been added to the rear wall.

»


sInglE-usE »

In many of these instances opportunities were taken to reduce both the complexity of operational set-ups and aseptic risk by incorporating multiple components in to one disposable solution that was provided as a single γ-irradiated assembly. The downside to this is that such customised solutions reduce supply chain flexibility, commonly taking 12 – 16weeks for order fulfilment and 2-4 weeks to complete design. Inclusion of these solutions should ideally represent a balance between the benefit provided by the customised solution and supply chain flexibility.

Product and Process ComparabilityFollowing completion of the first MAb drug substance manufacturing campaign in Q1 2009, process and product comparability between MacroGenics, Lilly BR&D (where numerous at scale developmental runs had also been performed), and the batches manufactured at Lilly Kinsale was assessed. Both the process and the product manufactured at Kinsale were deemed to be parametrically and analytically comparable to batches manufactured by MacroGenics and Lilly BR&D. The batches manufactured in Lilly Kinsale incorporated certain process changes (some described above), and inclusion of modified solution mixing parameters (following mixing studies and installation of standardised mixing systems).

The Lilly Kinsale site and API/MAb facility was inspected by the Irish Medicines Board in June 2009, with the agency granting a clinical trial manufacturing (IMPD) licence for the MAb process following a successful inspection outcome.

Concluding remarksThis article describes transfer, implementation and manufacture of Phase III drug substance material for a monoclonal antibody manufacturing process in a facility that was also employed for manufacture of a small molecule parenteral API. The MAb process is a largely fully disposable process, excepting affinity capture chromatography column and purification skids, which are cleaned and re-used.

Numerous considerations for transfer of single use MAb processes in to existing facilities were assessed, with the most prominent considerations being supply chain and analytical testing.

The case described here shows that adaptation of existing clean room facilities, coupled with the flexibility inherent in single-use MAb processes, is both feasible and can have successful outcomes in terms of agency inspections. This approach represents an alternate route to biopharmaceutical manufacturing, creating options for leveraging existing facilities, as opposed to constructing dedicated biopharmaceutical manufacturing space.

AcknowledgmentsThe authors would like to acknowledge their many colleagues in Lilly Kinsale (Operations, Quality, Procurement & Warehousing, MS&T, and Analytical), in Lilly Bioprocess R&D, and at MacroGenics who contributed to the work described in this article.

Author BiographiesBrian Mullan is a Scientist/Tech Transfer Lead, Manufacturing Science and Technology (MS&T), with Eli Lilly & Co, Kinsale, Cork, Ireland. Since joining Lilly he has been responsible for late stage development and transfer of biopharmaceutical manufacturing processes to launch sites. He previously worked for Centocor (Johnson & Johnson) in Cork, Ireland, and with Sanofi Aventis in Paris, Toulouse, and New Jersey. Brian holds a BSc in Biochemistry from University College Galway, Ireland, a PhD in Viral Genetics/Cell Biology from University College Cork, Ireland.

Aidan Collins is a Scientist, MS&T, with Eli Lilly & Co, Kinsale, Cork, Ireland. At Lilly he has been responsible for implementation of new biopharmaceutical processes (protein purification) for pipeline and in–licensed products. He has previously worked for BioUetikon, Baxter S.A., Pfizer, and Amgen prior to joining Lilly in 2007. He holds a BSc in Biochemistry and Molecular Biology, and a Masters in Quality Assurance from Dublin Institute of Technology, Ireland.

Kristi Huntington is an MS&T Consultant with Eli Lilly & Co, Indianapolis, IN, USA. At Lilly she is currently responsible for new product introduction (NPI) for drug substance manufacturing. She has previously held roles as NPI lead and MS&T Manager at Eli Lilly & Co, Kinsale, Cork, Ireland, and Technical Services Representative, Eli Lilly & Co, Carolina, Puerto Rico, and Indianapolis, IN, since joining Lilly in 2000. She holds a BSc in Chemistry from Hiram College, Ohio, and an MSc in Chemistry from The Ohio State University.

Marie Murphy is a Microbiology and Virology Lead with Eli Lilly & Co, Kinsale, Cork, Ireland. At Lilly she is responsible for the delivery and implementation of product protection strategies including viral safety for biopharmaceutical manufacturing. She has previously worked for EiRx Therapeutics, Cork, Ireland, and for Schering Plough, Cork, Ireland. Marie holds a BSc in Microbiology from University College Cork, Ireland, and a PhD in Virology from Trinity College, Dublin, Ireland.

join over 1000 of your colleagues

Discovery

• Phage & Yeast Display• Engineering Antibodies• Antibody Optimization

expression

• Difficult to Express Proteins• Optimizing Protein Expression• Purifying Antibodies

analytical

• Characterization of Biotherapeutics• Protein Aggregation and Stability• Immunogenicity

antiboDies

• Antibodies for Cancer Therapy• Bispecific Antibodies• Antibody-Drug Conjugates

Organized by Cambridge Healthtech Institute250 First Avenue, Ste 300, Needham MA, 02494 PEGSummit.com

the essential protein engineering summit

Experience the future of biologics.

sheraton boston hotel | boston, ma

keynote speakers

Raimund Dutzler Ph.D., Professor, Biochemistry, University of Zurich

Allan Bradley Ph.D., FRS, Director Emeritus, Wellcome Trust Sanger Institute

Wayne Coco Ph.D., Biologics Research, Bayer Schering Pharma AG

Davinder Gill Ph.D., Vice President and Head, Global Biologics Technologies, Pfizer, Inc.

James D. Marks M.D., Ph.D., Professor and Chief of Anesthesia, San Francisco General Hospital; Vice Chairman, Anesthesia and Perioperative Care, UCSF

Lorenz M. Mayr Ph.D., Executive Director, Unit Head Biology, Protease Platform, Novartis Pharma AG

Janice Reichert Ph.D., Research Assistant Professor, Tufts Center for the Study of Drug Develop-ment, Tufts University School of Medicine

Roger A. Sabbadini Ph.D., Founder, Vice President & CSO, Lpath, Inc.; Professor Emeritus, Biology, San Diego State University

William R. Strohl Ph.D., Vice President, Biologics Research, Biotechnology Center of Excellence, Centocor

Douglas Cecchini Ph.D., Director, Technical Development, Biogen Idec, Inc.

»


FormulAtIon DEvEloPmEnt »

IntroductionIt is frequently reported that the percentage of drug candidates that are limited by poor solubility is increasing [1,2]. These poorly soluble compounds typically require enabling formulations, and this trend creates challenges for teams in discovery and development who must drive in-vivo exposures high for animal toxicology studies and deliver robust dosage forms for clinical evaluation. Many enabling technologies are available for the formulator to consider, including lipids, cosolvents, surfactants, nanoparticles, cyclodextrin complexes, amorphous solid dispersions, and others. The suitability of the particular formulation approach depends largely on the physicochemical properties of the active pharmaceutical ingredient (API). Amorphous solid dispersions (ASDs) are particularly attractive for many poorly soluble drug candidates because these formulations offer many of the advantages of more conventional solid oral dosage forms but they also provide faster dissolution rates and higher drug concentrations in the gastrointestinal milieu [3]. Further, typical excipients utilized in production of ASDs are commercially available and they have proven to be well tolerated in vivo. We have successfully employed ASD technology to drive high plasma exposures in toxicology studies as well as to deliver challenging molecules in clinical studies. In this article we will discuss approaches for preparing, screening, characterizing, and dosing ASDs in preclinical and early development.

methods of PreparationRotary EvaporationRotary evaporation is a desirable method for preparation of ASDs for early stage (pre-clinical efficacy/toxicology, Phase I) studies. This approach is fast, material sparing, relatively inexpensive, and readily available. Moreover, a wide range of batch sizes from mg to kg quantities may be prepared with high yield. ASD preparation by rotary evaporation is carried out by first dissolving the API and formulation components (polymers, surfactants) in a pharmaceutically acceptable solvent. Typical solids load in the solvent is 5% to 25% by weight, and this is generally dictated by API/polymer solubility. The solvent is then removed in a rotary evaporator using heat (typically 40 to 80 oC) and vacuum. Total time for solvent evaporation can range from minutes to hours.

Brian E. Padden, Ph.D., Jonathan M. Miller, Ph.D., Timothy Robbins, Ph.D., Philip D. Zocharski, Leena Prasad, Julie K. Spence, & Justin LaFountaineAbbott Laboratories, Abbott Park, ILemail: [email protected].

Amorphous Solid Dispersions as Enabling Formulations for Discovery and Early Development

improved bioavailability.softgel solutions.

SOLVE WATER SOLUBILITY CHALLENGESImprove the performance of your BCS II/IV compounds with our innovative softgel delivery solutions.

© 2

011

Cat

alen

t Pha

rma

Solu

tions

. All

right

s re

serv

ed.

DELIVER MORE APIsBring your challenging APIs to market with our innovative, non-gelatin Vegicaps‰ capsules, which can handle a wider range of fi lls and higher melting points.

SOLVE PERMEABILITY CHALLENGESIncrease the permeability of your BCS III/IV compounds with our unique fi lm coating technology and fi ll formulations.

Discover more solutions with Catalent. Call: + 1 866 720 3148 Email: [email protected] Visit: www.catalent.com DEVELOPMENT DELIVERY SUPPLY

Catalent is your catalyst for better bioavailability. Our team of 1,000+ development and formulation scientists have the talent and expertise to solve the most complex bioavailability and permeability challenges to help you deliver more effective treatments. With over 75 years of experience and a wide range of innovative drug delivery solutions, we can improve the performance of your products, from discovery to market and beyond. Catalent. More products. Better treatments. Reliably supplied.™

8759 Catalent Softgel APR 8.125x10.875.indd 1 1/20/11 4:18 PM

»



Because of the relatively long evaporation time, the compound must have adequate chemical stability in the solvent at elevated temperatures. Longer evaporation times may lead to physical instability, so care must be taken to avoid API crystallization during solvent removal. This issue can be mitigated to a large extent through optimization of the temperature, vacuum, rotation, and total solids load. After removal of the solvent, the resulting ASD is isolated, dried, and milled to the desired particle size. Secondary drying in a vacuum oven or tray dryer is often employed to remove any residual solvent that remains in the final ASD powder.

The polymers that may be employed for ASD preparation by rotary evaporation are limited to those that can be easily isolated in high yield after solvent removal (Table 1). Hydroxypropylmethyl cellulose (HPMC) based polymers are typically not amenable to rotary evaporation, as these polymers often result in a glassy film that is difficult to isolate.

While preparation of ASDs by rotary evaporation is ideal for the early stages (pre-clinical to Phase I) of drug development, it is not well suited for later stage development, manufacturing, and commercialization. The process is not readily scaleable beyond quantities on the order of 10 kg because solvent volumes become too large, leading to very long and unrealistic evaporation times. Therefore, bridging to larger scale spray drying or hot melt extrusion (HME) processes is required if the drug candidate progresses into later stage development.

Spray DryingSpray drying is another method that is commonly utilized for the preparation of ASDs of poorly soluble compounds [4]. The method is readily scaleable from gram-sized batches during discovery and early development to kg and metric ton quantities during later stage manufacturing and commercialization. The first step in the spray drying process is to prepare a feed solution of the API and formulation components (polymers, surfactants) in a pharmaceutically acceptable solvent. The total solids load in the feed solution is typically 5% to 25% by weight, and this is generally dictated by API/polymer solubility as well as viscosity of the solution. The feed solution is then pumped into a spray nozzle along with inert, hot (typically 60 to 100 oC) drying gas where it is atomized and sprayed into a drying chamber. The solvent quickly evaporates during this process, leaving behind spray dried dispersion particles. These particles are collected in a cyclone with attached baghouse filter. Secondary drying in a vacuum oven or tray dryer is often employed to remove any residual solvent that remains in the final ASD powder. Because solvent evaporation time is extremely fast (on the order of seconds), spray drying is particularly advantageous for preparing ASDs of compounds with poor thermal stability.

There is no limitation to the types of polymers that may be employed for preparation of ASDs by spray drying. In particular, spray drying enables the preparation of ASDs in HPMC based polymers, which is often difficult, if not impossible, to achieve using rotary evaporation or HME processes. Spray drying also offers the opportunity to optimize particle size and bulk powder properties through process parameter optimization and also through the type of spray nozzle (e.g. two-fluid, ultrasonic, rotary, and pressure nozzles) [4]. In general, particle size

increases with equipment scale, as a result of larger droplet sizes and longer drying residence times. This may present difficulties during development, as particle size of the spray dried powder is inherently changing as the formulation is scaled. This can be especially challenging during discovery and early development, because smaller batch sizes lead to inherently small particles (~10 μm) which can lead to issues with flow and compressibility during downstream processing. Issues with particle size can be largely mitigated via dry granulation of the spray dried powder, however this adds another relatively complicated unit operation to the overall process. Another challenge to employing spray drying during discovery and early stages of development is that the currently available lab scale spray dryers suffer from poor yield and generally cannot work on mg quantities of material

Hot Melt ExtrusionHot melt extrusion (HME) is the most widely used method of preparation for ASDs for commercial products, because it is particularly well suited for large scale manufacturing [5,6]. The preparation of ASDs by HME typically involves the use of twin screw extruders to mix multiple materials (API, polymer, surfactant) into a melt which is extruded through a die. The extrudate is then cooled and either shaped by calendaring or pelletized and milled to a desired particle size. The final milled extrudate is then typically blended with additional excipients and compressed. Direct shaping to a final dosage form is also possible with calendaring or injection molding technology. HME is advantageous for commercial manufacturing because it is a continuous and easily scalable process. Unlike rotary evaporation and spray-drying, HME does not require the use of organic solvents, thus it is a “green” process that reduces cost and alleviates safety/environmental concerns. Processing must be performed at temperatures above the Tg of the polymer and high enough for the API to either melt and/or dissolve into the polymer matrix. HME can be limited in the ability to process heat sensitive and/or high melting point drugs and it is generally not amenable for manufacturing small (mg to g) quantities needed in preclinical development.

selection of ExcipientsPolymers Polymers are critical components in ASDs because they act as carriers for the drug and they inhibit crystallization in both the dosage form and in-vivo. By remaining in an amorphous state during dissolution, the drug can achieve supersaturation and potentially greater absorption, when solubility is the limiting factor.

In addition to in vivo performance considerations, polymer properties such as the glass transition temperature (Tg), solubility in organic solvents, and hygroscopicity must be considered in order to make the ASD stable and manufacturable. The properties of some commonly used polymers for preparation of ASDs are summarized in Table 1. The polymer Tg is an important property to consider when preparing and selecting an ASD formulation. Polymers with higher Tg have less mobility, lending to better inhibition of drug

»


« FormulAtIon DEvEloPmEnt

crystallization. Additionally, the polymer Tg is particularly important for hot melt extrusion, as the process must be carried out above Tg to sufficiently mobilize the polymer. Organic solvent solubility of the polymer is a critical factor when manufacturing by rotary evaporation or spray drying to ensure that the polymer can be fully dissolved at the required concentration. The hygroscopicity of the polymer must also be considered, because an increase in moisture content can negatively affect physical and chemical stability, and proper packaging may be needed for ASDs composed of hygroscopic polymers.

SurfactantsSurfactants are often used as solubilizers or emulsifying agents in ASDs. Their primary purpose is to increase the apparent aqueous solubility and bioavailability of the drug. The properties of some common surfactants used in ASDs are listed in Table 2. As with polymers, solubility in organic solvents is an important consideration when preparing ASDs from solvent. In the case of hot melt extrusion, surfactants can have a plasticizing effect, which allows processing at lower temperatures.

Organic SolventsSolvents are necessary when preparing ASDs by rotary evaporation or spray drying. The properties of some common solvents used for ASD preparation are listed in Table 3. Solubility of the drug typically drives the solvent selection process, but all components should be completely dissolved to produce a homogeneous feed solution and a consistent final ASD powder. The solubility of the components in the chosen solvent must be high enough to manufacture at a reasonable throughput (typically > 5% weight of the total solids load). Water is often employed as a cosolvent for drugs (e.g. hydrates) that exhibit maximum solubility in a water-organic solvent mixture. The boiling point of the solvent is used as a guideline to set process temperatures in both rotary evaporation and spray drying processes. Sometimes a system with multiple organic solvents can be used to improve the solubility of various components. For GLP/GMP manufacturing, the ICH limit of the chosen solvent must be considered and secondary drying is often necessary to remove residual solvent.

Table 1: Properties of Polymers Commonly Used in ASDs [7]

Polymer Tg (°C) Solvent Solubility Hygroscopicity Amenable Methods of Manufacture

Copovidone 106

Dichloromethane Ethanol

Methanol Water

Acetone

<10% @ 50% RHRotary Evaporation

Spray Drying Hot Melt Extrusion

Polyvinyl caprolactam-

polyvinyl acetate-polyethyleneglycol

copolymer

70Water

Ethanol Methanol Acetone

~5% @ 50% RHRotary Evaporation


PVP130 (K17) 168 (K30)

Chloroform Ethanol

Methanol Water

Acetone

~15% @ 50%RHRotary Evaporation


HPMC 170

Cold Water Dichloromethane: Ethanol

Dichloromethane: MethanolWater: Alcohol

<10% @ 50% RH Spray Drying

HPMC P 133 - 137Acetone: MethanolAcetone: Ethanol

Methanol: Dichloromethane2 – 5% @ 50%RH Spray Drying

HPMC AS 110 - 130Acetone*

Ethanol:Dichloromethane**clear or turbid viscous solution

~3% @ 50%RH Spray Drying

Methacrylate/methacrylic acid

copolymer 110 - 150

Ethanol, Methanol, Acetone, Acetone with 3% water

<5% @ 50% RHRotary Evaporation, Spray Drying, Hot Melt Extrusion

• Salt/polymorph/cocrystalscreening• Preformulationstudies• Amorphoussoliddispersiondevelopment• Excipientcompatibilityscreens• APIformselectionandoptimization• SolidstateNMRservice• Chiralresolutionbycrystallization• Solidstatemethodsdevelopment,validationandtransfer

BasedinChina,partneringwithpharmaceuticalcompaniestoprovidehighqualityintegratedsolidstatechemistrysolutionsforbothAPIandformulationdevelopment

High quality, fast turnaround, cost effective and flexible to suit all your needs

www.crystalpharmatech.com

Our Services

»



Small-Scale ASD ScreeningWhen conducting a screen for ASDs, the primary objective is to identify a formulation that enables in-vivo exposure of a poorly water-soluble compound and one that is also stable, both chemically and physically. For this, a wide range of polymers and polymer-surfactant systems can be screened. In addition, the drug and surfactant loading in the ASD can be evaluated for its effects on release behavior and in-vivo performance, as well as physical and chemical stability. If multiple combinations of polymers, surfactants, and drug/surfactant loads are screened, the number of samples can easily range into the hundreds.

For early stage screening, a centrifugal solvent evaporator can be used to quickly prepare a large number of samples, in parallel, using mg quantities of material. Samples can be prepared using 96-well plates for small (mg) scale screening or gram-scale quantities can be prepared using scintillation vials or small beakers [9]. Small samples allow for larger, more comprehensive screens to be carried out quickly while still providing enough material for meaningful characterization. Potential lead formulations can then be manufactured on a larger scale for further evaluation, including physical and chemical stability studies, in-vitro release characterization, and in-vivo studies in animals.

CharacterizationCharacterization of an ASD is a critical requirement following preparation in order to be confident in the performance of the formulation. Characterization should include analyses of both solid form and in-vitro API release in aqueous media. Numerous methods are available for characterization, the more common of which are described in the following sections and in Table 4.

Solid Form EvaluationOf primary importance is the identification of residual crystalline character as this signals potential long-term physical instability of the amorphous API. Polarized light microscopy (PLM) and powder X-ray powder diffraction (PXRD) are rapid, non-destructive techniques employed as an early screen for crystallinity. Individual crystals may be identified via microscopy while PXRD diffractograms provide information regarding the gross amorphous or crystalline character of the solid. Data from both methods provide greater insight when combined with more sensitive thermal methods such as differential scanning calorimetry (DSC).

DSC is employed to study transitions in the solid state as a function of temperature. Most often, ASDs are evaluated for the presence of melt endotherms and Tg. Observation of melt endotherms confirms the presence of crystalline components. Tg is an important descriptor that can provide insight into the long term stability of the ASD. Since a higher Tg generally indicates better stability, a single, high Tg value is desired for a particular ASD. Tg generally decreases with increased moisture uptake, which makes the API more prone to crystallization at higher RH conditions. Multiple Tg values indicates heterogeneity in the system and an increased potential for phase separation within the solid leading to crystallization. Thermogravimetric Analysis (TGA) is a thermal method that is complementary to DSC. TGA is employed to study weight loss from an ASD as a function of temperature and this technique is typically employed to roughly quantitate amounts of residual water or organic solvent present in the ASD. It is possible to determine the identity of materials lost during heating by coupling the TGA to a mass spectrometer (TGA-MS). These data may then be utilized to adjust processing or storage parameters to minimize the amount of plasticizing solvents which may lead to physical instability for the ASD. Dynamic vapor sorption can also be performed to understand the hygroscopic tendencies of the ASD powder.

Many other analytical methodologies are available to further characterize ASDs, although these are typically less common. Some of the other techniques include solid-state NMR spectroscopy [10], Raman spectroscopy [11], infrared spectroscopy [12], and isothermal microcalorimetry [13].

Table 4: Common Techniques Employed for ASD Solid Form Characterization

Technique Output Observations Analysis

Microscopy Photmicrograph BirefringenceCrystalline or amorphous

character based on presence or absence of birefringenge

PXRDX-Ray

Diffractogram

Presence or Absence of Observable, Intense

Peaks

Crystalline or amorphous based on characteristic presence or absence of

well defined reflections in diffractogram

TGA Thermogram Weight LossResidual water or organic

solvent

DSC Thermogram

Tg, singleHomogeneous amorphous

phase

Tg, multiple Phase Separation

Endothermic Events Desolvation or Melting

Exothermic Events Potential recrystallization

Table 2. Properties of Surfactants Commonly Used in ASDs [7]

Surfactant HLB Tm (°C) Solvent Solubility

Vitamin E Polyethylene Glycol Succinate 13.2 37 - 41Water

Acetone

Sorbitan Monostearate – 60/80 4.3 53 - 57 Most organic solvents

Polysorbate 20 16.7 n/aEthanol Water

Polysorbate 80 15.0 n/aEthanol Water

Polyoxyl 40 Hydrogenated Castor Oil 14 - 16 30

Chloroform Ethanol Acetone

Water

Table 3. Properties of Organic Solvents Commonly Used for Preparation of ASDs [8]

Solvent Boiling Point (°C) Flash Point (°C) ICH Class

Ethanol 78 13 II

Methanol 65 12 III

Acetone 57 17 III

IPA 83 12 III

Tetrahydrafuran 66 -14 III

Dichloromethane 40 None II

Published 11/10 PATH0153R0

US Headquarters

Patheon Inc. PO Box 110145 Research Triangle Park, NC 27709-5145 USA P: +1 919 226 3200 F: +1 919 474 2269 www.patheon.com

European Headquarters

Patheon International AG Lindenstrasse 14 6340 Baar Switzerland P: +41 41 766 2580 F: +41 41 766 2581 www.patheon.com

Why limit your compound to just one bioavailability enhancement technology?

One price, multiple technologies – fast results.

Call +1 866-PATHEON (+1 866-728-4366) or email [email protected]

• the first fixed-price, multi-platform scalable solution to increased bioavailability • parallel drug formulation screening including expert scientific analysis• SoluPath — the fast path to phase I trials and commercialization

»



Long term stability of ASDs should be evaluated due to the increased risk of both physical and chemical instability associated with amorphous solids. Accelerated stability studies should be conducted under stressed conditions (e.g. open dish at 40°C/75% RH, 60°C/75% RH, etc.) to understand both the physical stability of the ASD and any increased risk of chemical reactivity in the presence of excipients. Photostability should also be examined to determine whether special packaging is required to prevent photochemical reactions in the amorphous state. Understanding the stability of an ASD allows one to reformulate as necessary by varying drug load, by adding antioxidants, or by selecting alternative polymers/surfactants to maximize stability.

In-vitro ReleaseRelease of API from an ASD may be studied by a variety of in-vitro methods. One way to evaluate release from ASDs is by simple powder dissolution. This is performed by transferring a known mass of material into a known volume of biologically relevant dissolution medium (e.g. simulated gastric/intestinal fluid) under constant stirring. Solution concentrations are measured as a function of time to generate a concentration versus time release profile for the API. Solution concentrations in significant excess of the equilibrium API solubility in the particular medium should be targeted to test the ability of the ASD to achieve and maintain supersaturation.

However, it is desirable to be as biorelevant as possible to predict in-vivo performance. More dynamic techniques, such as the artificial stomach duodenum, and other multicompartmental dissolution/release methods, should be employed to understand API release from ASDs [3,14]. These methods are used to understand complex release phenomena and to begin building in-vitro/in-vivo correlations. A dynamic dilution scheme reported by Gao et al, was recently shown to provide valuable inputs for predictions of pharmacokinetics (PK).

Dosing ConsiderationsSince ASDs are inherently metastable systems, it is important to consider the implications of the dosage form and dosing conditions on physical/chemical stability, manufacturability, and in-vivo performance. Aqueous suspensions enable dosing of higher ASD amounts and they are therefore ideal for dosing to animals for toxicological evaluation. Care must be taken to ensure that the ASD does not crystallize in the aqueous suspension during preparation and dosing, which could compromise in-vivo performance. Gelling and/or foaming can occur when suspending an ASD in aqueous solution. This issue may be overcome through optimization of the ASD loading in the suspension and/or addition of anti-gelling/foaming agents. When dosing smaller animals by oral gavage, the particle size of the ASD in aqueous suspension must be small enough to pass through the gavage tube.

Hard gelatin capsules (HGC) are ideal for dosing of the dry ASD to larger animals and humans. If long term stability is required, the physical and chemical compatibility of the ASD with the capsule shell must be considered, especially for hygroscopic polymers which may dry out or cause the HGC to swell depending on the RH conditions.

Tablets are generally the preferred dosage form for ASD formulations of commercial products. Formulation of the ASD intermediate with secondary excipients (e.g. fillers, disintegrants, lubricants, glidants) is typically required in order to optimize the flow and compressibility of the ASD formulation to enable manufacturability of the finished tablet. Thus, the ASD intermediate must be physically and chemically compatible with the API/ ASD powder. The particle size of the ASD powder is also an important consideration for tablet formulations, as the particle size can affect the manufacturability (i.e. flow and compressibility) as well as the dissolution/release rate of the API from the tablet matrix. ASDs made by HME or rotary evaporation processes are typically milled to the optimal particle size for tableting. The desired particle size of spray dried ASDs may be obtained through process optimization, especially on large scale spray dryers. Spray dried powders made on a smaller scale during early development tend to have inherently small particle size, thus these powders may require dry granulation (roller compaction, milling) in order to obtain the desired particle size for optimal tablet properties.

ConclusionsWe have outlined the fundamentals of preparing, screening, characterizing, and dosing amorphous solid dispersions. Using these methods we have successfully delivered poorly soluble drug candidates for both preclinical and clinical studies and we believe that the use of ASD technologies will continue to increase. As the science of this drug delivery approach evolves and as new excipients become available, formulators will be able to design ASD systems that are even more sophisticated and that will ultimately get new medicines to patients faster.

references1. C. J. Lipinski, Pharmacol. Toxicol. Methods, “Drug-like properties and

the causes of poor solubility and poor permeability”,2000, 44, 235-249.

2. E. H. Kerns, J. Pharm. Sci.,“High-throughput physicochemical profiling for drug discovery“, 2001, 90, 1838-1858.

3. Gao, Y., Carr, R.A., Spence, J.K., Wang, W.W., Turner, T.M.; Lipari, J.M.; Miller, J.M., “pH-Dilution Method for Estimation of Biorelevant Drug Solubility along the Gastrointestinal Tract: Application to Physiologically Based Pharmacokinetic Modeling”, Molecular Pharmaceutics, in press.

4. Dobry, D.E., Settell, D.M., Baumann, J.M., Ray, R.J., Graham, L.J., and Beyerinck, R.A., “A Model-Based Methodology for Spray-Drying Process Development”, Pharm Innov. 2009, 4, 133–142.

5. Breitenbach, J.; Maegerlein, M., “Melt-extruded molecular dispersions”, Drugs and the Pharmaceutical Sciences, 2003, 133, 245-260.

6. Breitenbach, J., “Melt extrusion can bring new benefits to HIV therapy: the example of Kaletra tablets”, American Journal of Drug Delivery, 2006, 4, 61-64.

7. Handbook of Pharmaceutical Excipients, 6th Edition, Edited by Rowe R.C., Sheskey, P.J., and Quinn, M.E., Pharmaceutical Press, 2009.

8. Miller J.M., Blackburn A.C., Macikenas D., Collman B.M., and Rodríguez-Hornedo N., “Solvent Systems for Crystallization and Polymorph Selection”, in Solvent Systems and Their Selection in Pharmaceutics and Biopharmaceutics, AAPS Biotechnology: Pharmaceutical Aspects, Volume 6, 2007.

»


« FormulAtIon DEvEloPmEnt

9. Shanbhag, A., Rabel, S., Nauka, E., Casadevall, G., Shivanand, P., Eichenbaum, G., Mansky, P., “Method for screening of solid dispersion formulations of low-solubility compounds-Miniaturization and automation of solvent casting and dissolution testing” International Journal of Pharmaceutics, 2008, 351, 209-218.

10. Lubach, J.W., Munson, E.J., “Solid-State NMR Spectroscopy”, in Polymorphism: in the Pharmaceutical Industry, Edited by R. Hilfkiker, 2006, Wiley-VCH, 81-93.

11. Breitenbach, J., Schrof, W., Neumann, J., “Confocal Raman-Spectroscopy: analytical approach to solid dipsersions and mapping of drugs“, Pharm. Res., 1999, 16, 1109-1113.

12. Broman, E., Khoo, C., Taylor, L.S., “A comparison of alternative polymer excipients and processing methods for making solid dispersions of a poorly water soluble drug”, Int. J. Pharm., 2001, 222, 139-151.

13. Sebhatu, T., Angberg, M., Ahlneck, C., “Assessment of the degree of disorder in crystalline solids by isothermal microcalorimetry”, Int. J. Pharm., 1994, 104, 135-144.

14. Alonzo, D.E., Zhang, G.G.Z., Zhou, D., Gao, Y., Taylor, L.S., “Understanding the Behavior of Amorphous Pharmaceutical Systems during Dissolution”, Pharmaceutical Research, 2010, 27, 608-618.

Author BiographiesBrian Padden is Section Manager of Pharmaceutics at Abbott Laboratories. He holds B.A. degrees in physics and chemistry from Saint Mary’s University (Winona, MN), and M.S. and Ph.D. degrees in chemistry from the University of Minnesota. Dr. Padden started his career at the Schering-Plough Research Institute, where he was responsible for solid-state method development, GMP validation, and technology transfer to international manufacturing sites. Some of the commercial products that he contributed to during that time include Claritin®, Clarinex®, Asmanex®, Nasonex®, Noxafil®, and Zetia®. Dr. Padden then served in positions of increasing responsibility in the areas of preformulation and discovery support. In 2006, he joined Abbott and in his current role he is responsible for advancing the pipeline through material-sparing characterization and enabling preclinical formulations, in the therapeutic areas of oncology, neuroscience, pain, and dyslipidemia. He has received many technical awards, including the Abbott President’s Award in 2009, and he is a certified Lean Six Sigma Black Belt. Dr. Padden has published and presented dozens of papers, posters, and invited talks, and he is also a board member of the NSF Center for Pharmaceutical Development.

Jonathan Miller is a Principal Scientist in the Global Formulation Sciences - Solids group at Abbott Laboratories. He is also Adjunct Assistant Professor in the Department of Pharmaceutical Sciences at the University of Michigan. He has over 12 years of industrial experience both at Pfizer and now Abbott Labs, building broad expertise in the areas of pre-formulation, solid form screening/selection, formulation, biopharmaceutics, and material science. He has authored over 20 scientific publications and is an inventor on more than 10 patents and patent applications. In his current role at Abbott, he is responsible for the development of enabling formulations for poorly soluble compounds including amorphous solid dispersion formulations, lipid based drug delivery systems, and nano-

particles. Dr. Miller holds a Ph.D. in Pharmaceutical Sciences and an M.S. in Pharmaceutical Engineering from the University of Michigan. He obtained his B.S. in Biochemistry from Bowling Green State University.

Timothy Robbins is the Operations Manager of the Chemical Pilot Plant at Abbott Laboratories. Dr. Robbins joined Abbott in 1993 after completing post-doctoral work at the University of California, Los Angeles. He has over 17 years of chemical research and scale-up experience. He has 12 scientific publications and is an inventor on more than 7 patents and patent applications. In his current role at Abbott, he is responsible for the running of the Chemical Pilot Plant and Kilo Lab. Prior to joining the Chemical Pilot Plant, Tim worked on a number of projects as a process research chemist within the API Process R&D organization. Dr. Robbins holds a Ph.D. in Organic Chemistry from the Unviversity of Nevada-Reno. He obtained his B.S. in Chemistry from Olivet Nazarene University.

Philip Zocharski is a Senior Scientist in the Pharmaceutics group at Abbott Laboratories. Over his 12 year career in the pharmaceutical industry, Philip has focused his research and development efforts in the areas of pre-formulation and biopharmaceutics as a colleague of Parke-Davis/Pfizer and Abbott Labs. In his current role, he applies biopharmaceutics principles and a wide array of drug delivery technologies including amorphous solid dispersions, nanoparticles, and lipids to address the specific needs of teams in early Discovery. Philip holds an M.S. in Pharmaceutical Sciences from the University of Michigan. He obtained his B.S. in Chemistry from Michigan State University.

Leena Prasad is a Scientist I in the Global Formulation Sciences – Solids group at Abbott Laboratories. She graduated in 2005 from the University of Illinois at Champaign/Urbana with a B.S. in Mechanical Engineering. She joined Abbott in July of 2007. Since joining Formulation Sciences, she has actively been involved with various projects requiring solubility enhancement technologies and has worked with the group’s solid dispersion research team.

Julie Spence is a Scientist I in the Pharmaceutics group at Abbott Laboratories. She earned her B.S. in Chemical Engineering in 2001 from the University of Michigan and her M.E. in Pharmaceutical Engineering in 2002, also from the University of Michigan. Prior to joining Abbott in 2007, Julie worked as a Scientist at the Pfizer Ann Arbor, MI labs. (2002 - 2007). Over the course of her career, Julie has gained expertise in the areas of physicochemical characterization, dermatological drug delivery, biopharmaceutics, and solid form screening/selection. Julie’s current responsibilities include developing enabling formulations for toxicology studies.

Justin Lafountaine is an Associate Scientist II in the Global Formulation Sciences – Solids group at Abbott Laboratories. He graduated in 2007 from the University of New South Wales with a B.S. in Nanotechnology. After graduation, Justin worked briefly at Pharmaform before joining the Global Formulation Sciences group in September of 2007. Since then, he has actively been involved with expanding the group’s solid dispersion capabilities and implementing Process Analytical Technology.

»


PFs / E&l »

IntroductionPrefilled glass syringes (PFS) are increasingly becoming a container of choice for storing and administering therapeutic protein products to patients [1]. The PFS is a convenient and reliable system for injection compared with the more traditional method of transferring, measuring, and delivering a dose from a vial containing liquid or reconstituted lyophilized powder. In addition, the PFS can deliver a controlled volume minimizing drug waste associated with vial overfills.

PFS is a primary container and its compatibility with the drug needs to be addressed for ensuring patient safety and drug quality [2, 3]. Safety of PFS is the responsibility of drug manufacturers and understanding PFS extractables/leachables is important for assessing syringe compatibility. Identification and quantitation of extracted/leached chemicals is critical for assessing toxicological and drug quality risks. A high extractable/leachable risk assessment may simply disqualify a syringe and eliminate the need to conduct further resource intensive qualifications assessments such as stability, particle formation, container closure integrity, break loose extrusion, and others. Understanding extractable/leachable contributes to the PFS knowledge space and may also serve to create collaboration opportunities with suppliers for reducing/eliminating leachables and improving PFS systems [4]. This paper describes extractables/leachables from syringes and its implication on biological products.

Extractables and leachablesExtractables are chemicals that migrate from the product-contact component into a solvent at accelerated conditions (such as heat, time, pH, ionic strength, organic solvent content). Leachable are chemicals that migrate from the product-contact component into a formulated drug during normal storage/usage conditions.

Information regarding leached chemicals from components is not known or readily available from the supplier. Goals of extractable studies are to generate, identify, and predict leachables. Extraction under appropriate solvent, temperature, and exposure conditions can generate representative leachables and in large enough quantities to facilitate structure elucidation analysis [5, 6]. Assembled syringes and individual components can be extracted with various solvents (water and water/organic mixtures, pH, ionic strength) at elevated

Yasser Nashed-Samuel, Dengfeng Liu, Kiyoshi Fujimori, Lourdes Perez & Hans Lee, Ph.D.Department of Formulation and Analytical ResourcesAmgen Inc.

Extractable and Leachable Implications on Biological Products in Prefilled Syringes

Daikyo Crystal Zenith® and Flurotec® are registered trademarks of Daikyo Seiko, Ltd. Crystal Zenith® and Flurotec® technologies are licensed from Daikyo Seiko, Ltd. West and the diamond logo are trademarks of West Pharmaceutical Services, Inc. in the United States and other jurisdictions.

Copyright ©2011 West Pharmaceutical Services, Inc.

Breakage. Functional performance. Aggregation. These issues and more can affect your product and thus your top and bottom line. Glass syringes may be at the heart of the problems.

Made of a proprietary cyclic olefin polymer, the Daikyo Crystal Zenith Insert Needle Prefillable Syringe System provides an innovative solution to the limitations of glass syringes, especially with sensitive biologic drug formulations.

System advantages include:

•Highbreak-resistance

•Highestqualityassuredby100% vision inspection

•Superiorfunctionalperformanceandbarrier protection with Flurotec® film

•Nosiliconeoilonthesyringe barrel or piston

•Notungstenoradhesive

•Consistentfunctionalitywhenusedin auto-injectorsystems

Advance Your Delivery System with Daikyo Crystal Zenith® Insert Needle Prefillable Syringe Technology

Minimize Risk. Maximize Value. Enhance Delivery.

#6001

Contact West today.NorthAmerica+1800-345-9800Europe +4924037960AsiaPacific +6568605879www.WestPFSsolutions.com

Get the free mobile app at:http://gettag.mobi

6001 1ml CZ Syringe PFS ad.indd 1 2/7/2011 11:43:17 AM

»


PFs / E&l »

temperatures for a specified duration [5-8]. The identified extractables provide insight on predicting leachables. Leachables can be identical or a subset (oxidation, derivative, etc.) of extractables, or can form adduct products with other leachables, excipients, and proteins.

Extractables information is beneficial since analyzing leachables directly in the formulated drug is challenging due to low leachable concentrations (ppm/ppb range) and matrix inference from protein and excipients. Also categorizing foreign chemicals in drug products as syringe component or drug process impurity related is inconclusive when analyzing leachables directly. Extractables data can determine the source of the leachables.

Analytical techniquesAnalytical characterization and quantitation techniques associated with organic and inorganic chemicals are commonly used to analyze extractable/leachables [9]. Gas chromatography mass spectrometry (GCMS), solid phase micro extraction (SPME)-GCMS, liquid chromatography mass spectrometry (LCMS), high performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR) spectroscopy techniques are used to analyze unknown organic extractable compounds. Inductively coupled plasma mass spectrometry (ICPMS) can be used to identify and quantitate inorganic metallic elements. Evaporative light scattering detection (ELSD) can be supplemental to HPLC and suitable for oligomers/polymers and non-chromophores molecule analysis.

PFs Components, leachables, and ImplicationsSources of leachables can be from the PFS components (glass barrel, stainless steel hypodermic needle, rubber needle shield, silicone oil lubricants, and fluoropolymer coated rubber plunger) and less obvious sources are from [8, 10] residues from processing tools [11-13] and additives for attaching the needle to the barrel [4]. The impact of leachables on therapeutic proteins can be related to aggregation, particle formation, and/or product quality issues such as reaction with the formulation or protein. Many of these negative attributes may not be observed in non PFS containers possibly due to short contact times during the uptake and delivery from vials using non PFS. However, the shelf-life for biological therapeutics in PFS can be greater than one year allowing leachables to accumulate and potentially interact with drug formulations [14].

The impact from leachables may not be universally observed across all formulated products in a pipeline since formulation factors (pH, ionic strength, surfactants, excipients, etc.) and/or protein attributes may vary.

glassType I borosilicate glass is commonly used to make prefilled syringes and contains various inorganic oxides such as boron, silicon, calcium, sodium, potassium, iron, and aluminum [15]. These inorganic oxides may not pose a direct toxicological risk but migration of glass components from ion-exchange, glass dissolution, pitting, stress, surface layer exfoliation, weathering, and/or erosion corrosion effects may lead to particle formation. [7,15].

rubber materials (Plunger and needle shield)The plunger and needle shield are composed of rubber related materials. The product contact surface of the plunger is laminated with a fluoropolymer type film and the needle shield is unlaminated. Contact between the formulated drug with the plunger and shield may result in leached rubber additives or bromine related compounds. Leached rubber agents have been proposed to cause adverse patient effects [10].

hypodermic needleThe attached hypodermic needle is made of stainless steel. Inorganic metals used in the stainless steel formulation such as Fe, Cr, Mn, Ni, and Mo may leach from the needle. Based on our experience, metals leach at low levels and are not a direct toxicological concern. However under specific conditions and concentrations, metals such as Mn may catalyze oxidization reactions on proteins [16].

AdhesiveUV activated adhesives composed of organic and oligmeric/polymeric materials are commonly used to bind the stainless steel hypodermic needle to the glass barrel [4]. Controlling the adhesive formulation, application, activation/curing, residues, and clean up processes are important steps for preventing/reducing these materials from coming in contact and leaching into the drug formulation.

While evaluating PFS models from various vendors, several organic, oligomeric, and acrylate related materials consistent to those used in the adhesive industry were extracted and characterized [17]. These adhesive residues did leach into various formulated protein products and did not appear to induce precipitation, aggregation, or particles. The impact on drug quality was a concern since acrylates are reactive toward protein under certain conditions [18]. Evaluation of a formulated protein stored in syringes containing endogenous levels of adhesives at 37°C for 45 days led to adduct formation. Irreversible addition of acrylates with the formulated protein was observed at multiple lysine, histidine, and N-terminus sites with 0.02% of amino acid sites being modified [19].

SOME SKILLS ARE ESSENTIALSOME PRODUCTS AS WELL

AGUETTANT has designed a unique polypropylene prefilled syringe to improve safety and quality of care whilst optimizing cost levels.

Ready to use • Prefilled, preassembled and presented

in a blister pack • Sterility guaranteed inside & outside

through terminal sterilization

Integrity • Guaranteed by the frangible obturator

Tamper evident • Patented opening system with

pre-perforated label

Needle free • Secure connection with its universal

Luer Lock

INNOVATION FOR ALL

FOR YOUR PREFILLED DEVELOPMENTCome and visit us at BIOMEDevice on booth 345, Boston Convention & Exhibition Center, April 6-7th, 2011

AGUETTANT - 1 Rue Alexandre Fleming - 69007 LYON - France • www.aguettant.comTél: +33 (0)4 78 61 51 41 - Fax: +33 (0)4 78 61 09 35 - Email : [email protected]

»


PFs / E&l »

silicone oilSilicone oil is applied to coat the barrel, plunger, and needle exterior. The most common form of silicone oil used in medical applications is polydimethylsiloxane and functions as a lubricant allowing the plunger to glide smoothly within the barrel to expel the drug. As the demand for prefilled syringe and automated injection devices increases, so does the importance of understanding silicone oil. Functionally, silicone oil is not a major concern for manual injections since a nurse or doctor is capable of applying the necessary force to push the plunger to the end point. However, a spring can only provide a fixed amount of force and any unanticipated friction may cause the plunger to stall before complete drug delivery.

Although silicone oil is considered inert and insoluble in water, it may interact with the formulation causing protein aggregation [20,21], droplets, and particles [22]. Ideally the silicone oil application process should balance the need to minimize the undesired drug quality attributes and to provide sufficient lubrication. The silicone oil distribution is non-uniformly distributed with the least amount located near the needle end of the syringe (Figure 1) [23].

syringe tool – tungsten PinLess obvious sources of PFS extractables are contamination from tools that were used to manufacture and process syringes. Glass syringes are made from cutting and molding glass tubing at high temperatures. Heated glass is in contact with various processing tools during the process. The barrel’s inner channel cavity for holding a stainless steel needle is formed at approximately 1,200 °C using a tungsten pin. Tungsten is commonly used due its heat resistance and relative high melting temperature. However at temperatures greater than 150 °C, tungsten oxidizes in the presence of air and leaves a white tungsten containing residue within the syringe (Figure 2). These residues may survive the syringe washing step and may contact the drug upon syringe filling and storage. The tungsten oxide residues did not cause protein precipitation above pH 5, but caused protein aggregation below pH 5 [12, 24]. Below pH 5, ppm low amounts (parts per million, ppm) of tungsten oxide formed large tungstate polyanions which did aggregate protein at low ppm levels.

syringe tool – Polymeric PinPolymeric nylon pins are used to transport glass syringe barrels on an assembly line. These reusable pins (approximately 0.5 x 6 cm) fit within the hot syringe (Figure 3). Abnormal heat exposure or extensive pin usage may lead to pin wear and tear. Pin residues exposed to heated glass syringes may adhere to the inner syringe wall and survive wash and rinse procedures. During a visual inspection of a filled syringe product a black residue was observed (Figure 4) and later identified by LCMS and FTIR analysis as containing nylon related species [11]. Substances from the black residue did leach into the drug formulation

Figure 1. Silicone oil gradient in an empty PFS [23].

Figure 2. Tungsten oxides inside the syringe near the hypodermic needle-barrel zone [24].

Figure 3. Polymeric pin and a polymeric pin inserted into a syringe.

»


« PFs / E&l

and analysis confirmed the leachables and solid black residue matched those of the nylon pin used by the syringe manufacture. The syringe manufacturer was notified and has implemented measures to prevent future occurrences.

ConclusionPFS components and residues from processing tools may leach organic and inorganic chemicals into formulated drugs. Leachable information is often not readily available from the syringe manufacturer prompting drug manufacturers to initiate extractable/leachable investigations. Leachable from PFS may or have contributed to safety concerns related to protein aggregation, particle formation, and toxicological risk factors. Identifying extractables and leachables provide key information enabling safety assessments that address toxicology and drug quality impact for evaluating PFS.

AcknowledgementsAuthors would like to thank Joseph Phillips and David Brems for their efforts and useful discussions.

references1. Thompson, I. “New-Generation Auto-Injectors: Completing the

Scale of Convenience for Self-injection. Drug Delivery Report. 2005, Autumn/Winter, 47-49.

2. Markovic I. Risk Management Strategies for Safety Qualification of Extractable and Leachable Substances in Therapeutic Biologic Protein Products. Am Pharm. Rev. 2009, 12(4) 96-101.

3. Guidance for Industry. 1999. Container closure systems for packaging human drugs and biologics. Rockville, MD: US Department of Health and Human Services, Food and Drug Administration.

4. Sardella, A. Fine tuning of process parameters for improving biocompatibility of prefillable syringes. Ondrugdelivery. 2010, (January) 18-22.

5. Jenke, D. R. Evaluation of model solvent systems for assessing the accumulation of container extractables in drug formulations. J. Pharm. Sci. 2001, 224 (1-2), 51–60.

6. Jenke, D. R. Linking extractables and leachables in container/closure applications. PDA J. Pharm. Sci. Technol. 2005, 59 (4), 265–281.

7. Borchert, S.J.; Ryan, M.M.; Davidson, R.L.; Speed, W. Accelerated extractable studies of borosilicate glass containers. J. Parenter. Sci. and Technol. 1989, 43(2) 67-79.

8. Wakankar, A.A.; Wang, J. Y.; Canova-Davis, E.; Ma S.; Schmalzing, D.; Grieco, J.; Milby, T.; Reynolds, T.; Mazzarella, K.; Hoff, E.; Gomez, S.; Martin-Moe, S. On Developing a Process for Conducting Extractable-Leachable Assessment of Components Used for Storage of Biopharmaceuticals. J. Pharm. Sci. 2010, 99(5), 2209–2218.

9. Wang, Q. Selection of Analytical techniques for pharmaceutical leachables studies. Am Pharm. Rev. 2005, 8(6) 38-44.

10. Jenke, D. Suitability-for-Use Consideration for Prefilled Syringes. Pharm. Technol. 2008, April 1 issue.

11. Nashed, Y.; Torraca, G.; Liu, D.; Fujimori, K.; , Zhang, Z.; Wen, Z.; Lee, H. “Identification of an Extraneous Black Particle in a Glass Syringe: Extractables/Leachables Case Study.” PDA J. Pharm. Sci. Tech. 2010, 64, 242-248.

12. Jiang, Y. ; Nashed-Samuel, Y. ; Li, C. ; Liu, W. ; Pollastrini, J. ; Mallard, D. ; Wen, ZQ. ; Fujimori. K. ; Pallitto, M. ; Donahue, L . ; Chu, G. ; Torraca, G. ; Vance, A. ; Mire-Sluis, T. ; Freund, E. ; Davis, J. ; Narhi, L. J. Pharm Sci. 2009, 98(12), 4695-710.

13. Liu, W.; Swift, R.; Torraca, G. ; Nashed-Samuel, Y. ; Wen, ZQ. ; Jiang, Y. ; Vance, A. ; Mire-Sluis, A. ; Freund, E. ; Davis, J. ; Narhi, L. PDA J. Pharm. Sci. Tech. 2010, 64(1), 11-19.

14. Hung G.W.; Nunez L.J.; Autian J. Correlation of kinetic parameters and thermal behavior of segmented polyurethane elastomers with biological responses. J. Pharm. Sci. 1975, 64, 1492–1497.

15. Walther, M.; Rupertus, V.; Seemann, C.; Brecht, J.; Hormes, R.; Swift, R.W. Pharmaceutical Vials with Extremely High Chemical Inertness. PDA J. Pharm. Sci. Technol. 2002, 56(3), 124-129.

16. Deman, J. M. Principles of food chemistry 3rd edition. (1999) Aspen Publishers. ISBN: 0-8342-1234-X, 131-132.

17. Nashed-Samuel, Y. Extractable and Leachable Implications on Biological Products in Prefilled Syringes. PDA/FDA Joint Regulatory Conference, September 13-16, 2010.

18. Potter, D. W.; Tran, T. Rates of Ethyl Acrylate Binding to Glutathione and Protein. Toxicology Letters. 1992, 62, 275-285.

19. Liu, D.; Nashed-Samuel, Y.; Bondarenko, P. V.; Brems, D. N.; Ren, D. Interactions Between Therapeutic Proteins and Acrylic Acid Leachate. In preparation.

20. Jones, L. S., Kaufmann, A., and Middaugh, C. R. Silicone Oil Induced Aggregation of Proteins. J. Pharm. Sci. 2005, 94(4), 918-927.

21. Thirumangalathu, R., Krishnan, S., Ricci, M.S., Brems, D.N., Randolph, T.W., and Carpenter, J. F. Silicone Oil- and Agitation-Induced Aggregation of a Monoclonal Antibody in Aqueous Solution. J. Pharm. Sci. 2009, 98(9), 3167-3181.

22. Markovic, I. Challenges Associated with Extractable and/or Leachable Substances in Therapeutic Biologic Protein Products. Am. Pharm. Rev. 2006, 9(6), 20-27.

23. Lee, H.; Liu, D.; Fujimori, K.; Perez, L.; Nashed-Samuel, Y. Unpublished results. 2006.

Figure 4. Two black particles observed inside a pre-filled syringe. Larger particle is approximately 300 microns [11].


PFs / E&l »»

24. Lee, H.; Nashed-Samuel, Y.; Fujimori, K.; Liu, D.; Perez, L. Tungsten Leaching from Prefilled Syringes and Impact on Protein Aggregation. Poster presented at the PDA Extractables/Leachables Forum: Confronting Extractables and Leachables Issues in an Evolving Industry; Bethesda, Maryland, November 6–8, 2007.

BiographyYasser Nashed-Samuel, Ph.D., is currently a principal scientist at Amgen (Thousand Oaks, CA), Process and Product Development, R & D organization. Since joining Amgen in 2003, he established and led the leachables and extractables (L/E) effort. The L/E group engages in assessing product contact for manufacturing equipment, bulk containers, primary delivery containers and devices and incident investigations for both clinical and commercial products.

Dengfeng Liu, Ph.D. is a senior scientist in Process & Product Development at Amgen. He joined Amgen in 2006. Dengfeng leads structure elucidation for L/E by mass spectrometry and NMR spectroscopy. He conducted research on the interactions between small molecules therapeutic proteins.

Kiyoshi Fujimori. Graduated University of California, Los Angeles in 1996 with BS in biochemistry. Initially worked as peptide chemist at Bachem for six years. Joined Amgen in 2004 and currently studying leachable and extractable as associate scientist in Product Contact Assessment team of Process and Product Development.

Lourdes Perez is currently a Sr. Associate at Amgen (Thousand Oaks, CA), Process and Product Development, R & D organization. Since joining Amgen in 2006, she has supported leachables and extractables (L/E) projects, incident investigations, and primary delivery containers and devices utilized for clinical and commercial products.

Hans Lee, Ph. D. has been with Amgen since 2003 in the Process and Product Development organization and is responsible for extractables/leachables activities related to assessing product contact for manufacturing/infusion/device equipment and bulk/primary containers for clinical and commercial products. Hans has a Ph.D. in inorganic chemistry at UCLA.

300+

CHARLOTTE’S PREMIERBOUTIQUE HOTEL

MODERN GUESTROOMS

MEETING & EVENT ROOMSCENTRALLY

LOCAT

ED IN

DOWNTO

WN

CHARLO

TTE

6200# OF GUESTS OURMEETING SPACE CAN ACCOMMODATE

555 South McDowell StreetCharlotte, NC 28204

RESERVATIONS: 888-664-6835blakehotelnc.com

THE

FITNESSCENTER

What is your company’s primary business? (Check one only)1 Pharmaceutical Manufacturing 8 Bulk Materials/Nutritionals

2 Biopharmaceutical Manufacturing 9 Specialty Ingredients/API

3 Contract Pharmaceutical Manufacturing 10 Government

4 Medical Device/Instrument Manufacturing 11 College/University

5 Clinical Pharmaceutical Services 12 Contract Labs

6 Contract Pharmaceutical Research 99 Other (please specify below)

7 Pharmaceutical Engineering/Consultant Services

What is your job function? (Check one only)A Research & Development E Lab Management J Purchasing

B Quality Control/Assurance F Project Management K Engineering

C Validation G IT/Data Management L Corporate Management

D Production Manufacturing H Regulatory Affairs Z Other (please specify below)

Areas of Interest? (Check all that apply)1 Analytical Methods 12 Injectables 23 Process Control

2 Aseptic Processing 13 Lipids 24 Single Use Technology

3 Bioprocessing 14 Lyophilization 25 Solid Dosage Formulations

4 Chirality 15 Microbiology 26 Thermal Analysis

5 Controlled Release 16 Molecular Imaging 27 Validation

6 Dissolution 17 NIR 28 Liquid Chromatography

7 Drug Delivery 18 Parenteral Manufacturing 29 Mass Spectroscopy

8 Excipients 19 Particle Sizing 30 Raman

9 Extractables/Leachables 20 PAT/QBD 31 SFC

10 Filtration 21 Peptides 99 Other (please specify below)

11 HPLC 22 Pharma IT

Which tradeshows/conferences do you plan to attend? (Check all that apply)A PITTCON E INTERPHEX Puerto Rico J Joint Molecular Imaging Conference/WMIC

B EAS F Controlled Release Symposium K PDA

C IFPAC G HPLC L ISPE

D INTERPHEX H FACSS M ASMS

N AAPS

All of the above questions must be answered to qualify for your free subscription. Free subscriptions are available within the United States and Canada. Readers in other countries may subscribe for an annual subscription at the rate of $135.00.

FAX form to (317) 816-8788 or for IMMEDIATE service, simply subscribe/renew at www.myamericanpharmaceuticalreview.com

I want to receive/renew American Pharmaceutical Review FREE YES NoWhich version of American Pharmaceutical Review would you like to receive? Print Digital (include your email address below)

I want to receive APR News in Review enewletters YES No

Signature ____________________________________________________________________________ Date __________________

Name ________________________________________________________ Job Title _______________________________________

Company _________________________________________________________________________________________________________

Address __________________________________________________________________________________________________________

City ______________________________________________________ State ____________ Zip _______________________________

Phone _______________________ Fax _________________________ Email ______________________________________________

BW1X1B

subscription& RENEWAL

»


EXCIPIEnts »

Irwin B. Silverstein, Ph.D. Vice President and Chief Operating Officer, IPEAemail: [email protected]

Selling the Audit Observation to Enhance Conformance

IntroductionAudits are typically conducted to verify conformance to requirements and to identify gaps between current practices and desired outcomes. GMP audits are specifically conducted to assess regulatory compliance while quality audits identify opportunities to drive down cost and improve quality while maintaining regulatory compliance and customer satisfaction. It is generally easy to convince the auditee to implement cost improvement opportunities identified through quality audit since the impact on competitiveness, and thus their livelihood, is readily apparent. However when the GMP audit identifies a gap against regulation or desired outcome, or compendial specifications where applicable, there is often the perception that the improvement does not add value while raising costs and thus the improvement is often considered bureaucratic.

An apt comparison can be made with driving laws. When new drivers are informed of the rules that drivers are to follow, compliance is based upon passing the written and then the driving test followed by the financial consequences of the driver getting a ticket. However, when compliance is sold on the personal impact to driver safety, since compliance equates with avoiding an accident, following the rules of the road is expected to rise to a higher level. We see this implemented in public service advertising intended to link the affect of drinking to unsafe driving and more recently, texting to accidents.

In this article, I will demonstrate the value of establishing the linkage between failure to comply with regulation or specification and the potential impact to patient safety. The premise is that people would not want to knowingly jeopardize the safety of pharmaceuticals dispensed to themselves, family members, or acquaintances let alone to the general public.

Excipient GMPs are a guideline issued by the International Pharmaceutical Excipients Council jointly with the Pharmaceutical Quality Group. As such excipient GMPs are not regulation. The interpretation of conformance expectations are those of the author based upon his participation in the development of the initial guide in 1995 through its revisions to date.

In GMP audits of all too many excipient manufacturers’ quality control laboratories, I have observed insufficient documentation of test details. This is manifest in two aspects; first is the failure to maintain a permanent record of measurements particularly scale weights (gross and tare), and the second involving recording critical aspects of test sample preparation, e.g. time and temperature. While documentation of such details is not an explicit requirement for excipients, it is a practice taught in post-graduate science education. If you consider the impact the lack of such details has on the operation of the Quality

Go for RetaLac®.With MEGGLE’s new RetaLac®, a co-processed excipient, consisting of 50% Lactose Monohydrate and 50% HPMC you can compress HPMC directly with no problem at all. Thanks to its good flow properties and good wettability, sustained release tablets can be produced much more quickly, easily and efficiently (both for DC and wet granulation). What’s more, RetaLac® gives you maximum flexibility for your formulations. It enables the proportion of active ingredients to be as high as 60 %, and their release can be modified by adding further carriers.

RetaLac®, the world’s first Lactose-HPMC coprocessed excipient for direct com-pression, with good flow properties, good wettability and high flexibility.

WORLD FIRST!

MEGGLE USA Inc.50 Main Street, 10 th Floor

White Plains NY 10606, USAPhone: (914) 682 6891

Fax: (914) 682 6896E-mail: [email protected]

Mutchler Inc. Pharmaceutical Ingredients Harrington Park, NJ , USAPhone: 800 630 3440 Fax: (201) 768 9960 E-mail: [email protected] www.mutchlerchem.com

Meggle_Anzeigen_206x276.indd 5 27.01.11 09:37

»


EXCIPIEnts »

Control Laboratory, you quickly realize it greatly inhibits identification of mistakes and the investigation of out-of-specification test results. In order to justify retest or resample, it is beneficial to be able to document that the original test was improperly performed based on review of the test record. Otherwise the original result cannot be readily negated and the incident would have to be more thoroughly investigated as an out of specification test result.

While this argument would seem sufficient to motivate better documentation in the laboratory, nothing illustrates the importance of recordkeeping as an example. During a recent audit, the record for the preparation of a laboratory reagent solution was reviewed. The laboratory notebook merely listed the title of the reagent, the date, the preparer, and the quantity of water used. What was lacking was the weight of reagent used. Unfortunately, the title of the reagent solution recorded in the notebook indicated a 20% solution was prepared whereas the intent was to prepare a 15% solution. The investigation was limited to confirmation through testing that in fact the reagent was a 15% solution. Clearly it would have been far simpler to verify the quantity of reagent mixed with water would yield the intended 15% solution.

This example also illustrates other failures in the laboratory including a failure to perform an independent second check of the test record. However, without the additional test data being recorded, a second check would probably not have identified the discrepancy. Had the solution in fact been improperly prepared, there was a potential to release excipient unknowingly with an elevated impurity.

Laboratory records should not only record the weights and volume of materials used but also important details that demonstrate the test was performed in accordance with the proscribed test method. If such conditions as temperature, time, and the use of specified laboratory equipment are important to assuring proper sample preparation, than those details should also be recorded.

The next example in selling the audit observation involved the requirement to provide adequate toilet facilities. During the inspection of a bulk loading station, a portable toilet was observed. Upon questioning the escort, it was established that this facility was provided for use of the loader. This operation has a sole loader each shift and the loading station is staffed 24/7 365 days per year. Activity at the loading station was such that the loader was unable to leave for the time required to use the nearest available rest room.

A portable toilet provides a sanitary facility to relieve one’s self but generally does not provide for hand washing. When the lack of washing facilities was noted, the host rationalized that the soiled hands of the loader would not present a contamination risk to the excipient since the hands of the loader do not come into contact with the excipient. The auditor noted that the loader is required to take a dip sample from the truck or railcar into a glass bottle secured in a metal holder. The escort responded that they are required to wear gloves as a safety requirement during all loading operations and therefore there was no contamination risk. The auditor then pointed out that the loader would have an easier time putting the sample bottle into the holder without wearing bulky gloves and the escort agreed that this was likely the case. It is important to always remember that employees will take the easiest path. Thus it was established that the lack of a hand

washing facility presented a risk of contamination to the excipient during bulk loading.

The next area where selling the observation was beneficial to getting a commitment to improve the situation was in packaging. All lighting fixtures are expected to be protected against accidental breakage of the bulb. All light bulbs present a contamination risk due to the fine shards of glass their breakage can introduce into the material. Fluorescent bulbs present the added risk from the mercury they contain. Therefore it is prudent to protect bulbs from being struck or from falling from the fixture and shattering; particularly where excipient is packaged.

Unprotected fluorescent fixtures were in use in the area where excipient was being packaged into bags. The fixtures were located roughly 7 feet above the floor and about 5 feet from the feed nozzle at the bag packaging facility. The auditor noted the contamination risk should a bulb break and the escort rightly observed that neither glass nor mercury would likely get into the bag. However, the regulatory expectation is that the bulbs of such fixtures should be protected.

It was observed during packaging that the excipient bags were stacked below the subject lighting fixture. The auditor presented the argument that should a bulb break, the glass and mercury would fall onto the bags of packaged excipient. While the fine shards of glass would not likely penetrate the bag, it would be virtually impossible to remove all glass from the affected bags onto which the glass shards fell. It was pointed out that excipient packaged in bags is often introduced into the pharmaceutical manufacturing process by slitting the bag open and shaking out the contents. It was thus apparent that glass on the outside of the bag could very well be introduced into the process. Therefore the potential for glass contamination in the pharmaceutical product was evident and the manufacturer protected the light fixture.

It seems that many excipient manufacturers are reluctant to audit their suppliers. Yet oftentimes excipient manufacturers accept incoming raw materials solely on the basis of COA and some form of identity confirmation. This is acceptable practice as long as there is a sound basis for reliance on the supplier’s test data as reported on the COA. Excuses for not visiting the supplier include:

• The facility is dedicated to the manufacture of the material.

• The material is a commodity.

• I’ve used their material for quite some time without a problem.

• I don’t know what quality standard to audit against.

The last excuse is the simplest issue to address. The supplier audit should be conducted against ISO 9001 since conformance to this quality system requirement is the expectation of many non-pharmaceutical customers.

The remaining excuses would all seem reasonable but a visit to the supplier provides several benefits to the customer-supplier relationship. First it personalizes the relationship between the excipient manufacturer and their supplier. Second it educates the supplier to the excipient application for their material and by informing them the material is used to make a pharmaceutical ingredient; it should motivate the supplier to maintain their conformance to specification.


»« EXCIPIEnts

Finally, it offers an opportunity to assure the supplier is aware of the importance in notifying the excipient manufacturer of changes that may alter the composition or performance of the material in the customer application.

The final example of selling the audit finding deals with change control; referred to in the chemical industry as Management of Change (MOC). Excipient manufacturers who are part of the chemical industry generally operate under MOC due to the importance of safe operation of the facility. Any change that can potentially affect safety in the plant is carefully reviewed under MOC before being implemented. This facilitates addressing the issue of change control for excipient manufacture since adding a quality review to MOC assures the impact of change to the excipient is considered. While MOCs are carefully reviewed to assure the change is safe to implement, even where there is a quality review of the process change for its potential to affect the quality of the excipient; sometimes the assessment is not sufficiently rigorous.

The objective in selling the audit finding is to change the paradigm of the impact assessment of the change on excipient quality. A site installed new equipment to affect temperature control. While the new equipment was not a replacement in kind, an assessment of the new equipment would conclude that it was unlikely to impact the quality of the excipient. To address the remote possibility of product contamination, the MOC required running a fixed quantity of distilled product through the new equipment and collecting this material for reprocessing. The reprocessed material was to be distilled and sold.

While distillation is a good purification technique, the site never used distillation to assure the removal of trace impurities left over from metal fabrication. The new equipment might contain residues of process materials such as oils, used in its manufacture. The presence of such material in the product processed through the new equipment to demonstrate equipment performance and to assure the equipment was clean, can present a contamination risk. Excipient that contains such residue which is subsequently distilled might contain trace quantities of a new impurity or impurities. The discussion focused on the potential for this admittedly minimal risk to excipient quality until it was noted that the company does not to reprocess or rework any excipient grade material that has been rejected by the customer for any reason. This policy is attributable to the small risk of introducing impurities to the excipient from customer’s sampling the excipient or other activities by the customer. Once the site was reminded of this policy on customer returns, it was apparent to all that the plan to reprocess the purge from commissioning activities conflicted with the company policy on reprocessing material returned from the customer. This intellectual exercise has sensitized the site to a more rigorous assessment of MOC on excipient quality.

There are times when selling the observation is not apparent. The expectation is for all processing lines to be labeled. However, the auditor is often challenged to identify the benefit to the manufacturer for labeling the lines when the facility is dedicated to the manufacture of the excipient. Tracing the line to the source or destination unambiguously establishes the identity of the material container therein. The auditor is still searching for a tangible benefit where lines are fixed!

Selling the audit observation to the auditee is much more challenging than citing chapter and verse from the regulation or guideline. It often requires quick thinking on the part of the auditor but it can result in a paradigm shift in thinking at the audited facility. This intellectual exercise will continue to be the best means to convince the auditee to change their practices for improved conformance to excipient GMP until there are regulatory examples, i.e. FDA 483s or Warning Letters, to cite.

Author BiographyIrwin Silverstein is a consultant specializing in quality assurance and regulatory compliance for pharmaceutical excipient ingredients. He has worked since 1991 with the International Pharmaceutical Excipients Council (IPEC) developing guidance documents for excipient GMP compliance. He has been the VP and Chief Operating Officer of International Pharmaceutical Excipients Auditing Inc (IPEA) since 2001 where he was instrumental in developing the ANSI accredited IPEA Excipient GMP Conformance Certification program.

»


Our company is a global leader in mass spectrometry with a broad range of innovative instrument systems, software and services used to discover new drugs, advance medical science and protect the food supply and the environment. AB SCIEX solutions, including the AB SCIEX TripleTOFTM 5600 System for the fastest and most sensitive high-resolution mass spectrometer for qualitative and quantitative analysis, combine the highest performance with the highest reliability to enable scientists to fuel scientific discovery and deliver results with confidence.

For more information, go to our website.

Booth # 3535www.absciex.com

We are a leading manufacturer and supplier of specialty and high purity chemicals available in quantities for research or production. The Alfa Aesar Catalog includes more than 30,000 products and over 3,000 new items. In addition, the catalog also includes a full line of Platinum Labware, Spectroflux® alkali borate analytical fluxes

and the Specpure® brand of analytical standards.

Booth # 4945www.alfa.com

The Bruker name has become synonymous with the excellence, innovation, and quality that characterizes our comprehensive range of scientific instrumentation. Our solutions encompass a wide number of analytical techniques ranging from magnetic resonance to mass spectrometry, to optical and X-ray spectroscopy. These market and technology leading products are driving and facilitating many key application areas such as life science research, pharmaceutical analysis, applied analytical chemistry applications, materials research and nanotechnology, clinical research, molecular diagnostics, and homeland defense. Bruker — Innovation with

Integrity!

Booth #2561www.bruker.com

Our market leading IC systems redefined IC with RFIC, suppression, and online capabilities and range from basic to the worlds most advanced capillary IC. Our new UHPLC+ focused improvements make all UltiMate® 3000 systems UHPLC

capable, including the high performance RSLC, RSLCnano, BioLC, semiprep and standard. Our Chromeleon® software turns samples to results fast. Our advanced array of IC and LC column chemistries deliver unrivaled separations. Sample prep solutions, Accelerated Solvent Extraction (ASE®) systems and new AutoTrace® 280

SPE.

Booth # 2861www.dionex.com

Distek presents the ActiPix SDI300 dissolution imaging system with the unique capability of quantitative imaging of the liquid/surface interface for a diverse range of substances. Distek will also show their “bathless” and bath based Dissolution Systems along with a variety of products for automation including; Evolution 4300 autosampler and Opt Diss In-situ UV Fiber Optics. Visit Pittcon Booth #1960 to see

the NEW products and be entered to win an iPod touch

Booth # 1960www.distekinc.com

A global organization continuing their focus on technology strategies encompassing a wide array of Laboratory and Scientific instruments. Exhibiting a product line that covers particle sizing and Zeta Potential-analyzers using both dynamic and static light scattering, digital image analysis, optical microscopy and acoustic attenuation technology. Highest performance in spectroscopic instrumentation: Raman/PL microscopes with rapid imaging; spectrofluorometers; EDXRF microscopes; ICP; C/S, O/N & H elemental analyzers; InGaAs arrays, OEM miniature spectrometers & Raman systems & gratings. HORIBA remains committed

to global environmental conservation.

Booth # 1922, 1923www.horiba.com

The following pages list profile and booth information for advertisers that will be exhibiting at the 2011 Pittcon Conference & Expo.

»


A Rockwell Collins Company, Kaiser is recognized as a world leader in the design and production of Raman analyzers and components for in situ Raman spectroscopy. Kaiser’s suite of analyzers includes instruments for microscopy & imaging, reaction monitoring, gas-phase Raman, solids sampling, and transmission Raman. Raman analyzer installation locations include R&D, Pilot plant, manufacturing, and QA/QC. Application areas for RamanRxn Systems™ analyzers include the pharmaceutical, biotech, semiconductor, nanotechnology, petrochemical, polymer, and specialty chemical areas. Kaiser offers a range of Raman probes and optics to meet your

sampling needs.

Booth # 1648www.kosi.com

Our company has become synonymous with expertise in weighing and analysis instrumentation for laboratories. The laboratory division manufactures and markets a full range of precision products including balances, pipettes, titration equipment, thermal analysis instrumentation, density & refractive index determination equipment, moisture analyzers, and laboratory automation systems. METTLER TOLEDO products are fully supported by factory-trained service representatives

who perform calibration, qualification, and validation services.

Booth # 2726, 2727www.mt.com

Thermal analysis, calorimetry, thermal properties, & contract testing services; DSC, DTA, TGA, STA (Simultaneous DSC/DTA-TGA) from cryogenic to +2400C, evolved gas analysis by coupled FTIR & MS featuring a new TGA-GC-MS system, adiabatic reaction calorimeters (ARC & APTAC) to measure thermal & pressure properties of exothermic chemical reactions, new MMC 274 tabletop reaction calorimeter, thermal conductivity, thermal diffusivity by laser flash & xenon flash to +2800C,

DMA, TMA, DEA for in-situ thermoset cure monitoring, & more.

Booth # 3126www.netzsch-grinding.com/pharma

Products for HPLC/UHPLC sample prep and chromatrography applications are available from Pall Life Sciences, the leader in Filtration and Separation.

From the newly released Advance line of filter plates and centrifuge filters, to the industry leading Acrodisc PSF syringe filter, Pall continues to provide solutions that improve your processes and results. Stop by our booth #2353 to see a variety of products designed specifically for purificaiton, detection, sample prep and quality

control. For more information, visit Pall Life Sciences at www.pall.com/lab.

Booth # 2353www.pall.com/biopharm

Cutting-edge technology. Ultimate commitment. PANalytical designs, develops, and supplies X-ray analytical instrumentation and software solutions for materials characterization. Whether in the drive for comprehensive R&D solutions or superior quality control, PANalytical’s X-ray diffraction and X-ray fluorescence systems deliver quality analytical results. Please visit us to see the latest technological advancements in XRF, XRD and sample prep equipment, software, standards and quality programs, all delivered with the application expertise for complete

solutions to your material analysis challenges.

Booth # 2261www.panalytical.com

PSS is a major force in developing particle size analyzers for both wet/dry applications. Nicomp DLS (0.5nm-6 microns) offers nano sizing while AccuSizer SPOS (0.15-400+ microns) offers a wide dynamic size range providing high resolution, high sensitivity accurate particle size information. The AccuSizer FX PAT. offers high concentration SPOS that has the sensitivity to detect small differences between particle size distributions. Our product line is rounded out by high resolution image analysis and Archimedes SMR technology, an ultra-high resolution mass sensor that weighs each particle, providing submicron counting

measurements of mass and size.

Booth # 1116www.pssnicomp.com

»


Phenomenex is a global technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers in industrial, clinical, government and academic laboratories. From drug discovery and pharmaceutical development to disease diagnosis, food safety and environmental analysis, Phenomenex chromatography solutions accelerate

science and help researchers improve global health and well-being.

Booth # 4634www.phenomenex.com

We are a leader in providing automated dissolution testing systems, content uniformity and assay workstations and automated physical tablet testing instruments for the pharmaceutical, medical device, biopharmaceutical and dietary supplement industries. New for 2011 are JT Baker’s Dilut-IT media concentrates, direct HPLC or UPLC analysis and UV Fiber Optic Dissolution. Learn

how to automate your pharmaceutical testing on our website.

Booth # 1447www.sotax.com

Visit our exhibit and see world’s largest portfolio anywhere including analytical instruments, reagents, laboratory consumables, equipment, and services. Whether you need an instrument, an entire application workflow, or laboratory workstations, think Thermo Scientific. You’ll find Thermo Scientific innovation and the latest products to help you run your laboratory at peak performance and run

your experiments from start to finish. See the entire line up on our website.

Booth # 2835www.thermo.com

Our company helps laboratory-dependent organizations by providing breakthrough technologies and solutions. Pioneering a connected portfolio of separation and analytical science, laboratory informatics and mass spectrometry, Waters provides the tools to improve the quality of today’s science and explore the

infinite possibilities of tomorrow. Waters, The Science of What’s Possible.

Booth # 1635www.waters.com

Watson-Marlow Pumps, Tubing and Flexicon fillers are designed for pharmaceutical processing and laboratory applications for fluid transfer, metering, dispensing, and filling. These processes demand sterility and a high degree of precision to ensure the end product meets the industry’s strictly regulated quality standards. For pumping or filling aseptically, nothing beats Watson-Marlow products. Offering a contamination-free single use fluid path, Watson-Marlow’s peristaltic technology simplifies cleaning validation and enhances the integrity of high purity upstream

processes, purification, and fill/finish applications.

Booth # 764www.wmpg.com

WITec is a manufacturer of high resolution optical and scanning probe microscopy solutions for scientific and industrial applications. A modular product line allows the combination of different microscopy techniques such as Raman, NSOM or AFM in one single instrument for flexible analysis of optical, chemical and structural

properties of a sample.

Booth # 1420www.witec.de

A complete solution for your pharmaceutical

information needs.

9225 Priority Way West Drive, Suite 120, Indianapolis IN 46240317-816-8787 | Fax 317-816-8788

American Pharmaceutical Review is a peer reviewed journal

that reaches over 30,000 subscribers within the North American

pharmaceutical, biopharmaceutical and contract pharmaceutical

market. Each monthly issue contains editorial tracks on

Bioprocessing, Manufacturing, PharmaIT, Aseptic Processing,

Parenteral Manufacturing, and PAT.

Our website, www.AmericanPharmaceuticalReview.com, provides

subscribers with an online extension of our magazine. Focusing on

content, the site contains over 600 articles searchable through

our advance keyword search capability. In addition, we feature

industry news, events, and white papers.

Subscribe for free at

www.americanpharmaceuticalreview.com

APR_PuzzleFull.indd 1 12/10/10 2:04:59 PM

New Chromogenic Endotoxin Detection SystemAssociates of Cape Cod, Inc., (ACC) is proud to introduce its NEW CHROMOGENIC ENDOTOXIN DETECTION SYSTEM incorporating the enhanced Pyrochrome® chromogenic reagent and the new‐to-market Pyros Kinetix® Flex Incubating Kinetic tube reader. This system offers our customers diverse options for conducting endotoxin testing.

ACC’s enhanced Pyrochrome® is a versatile, quantitative reagent for performing kinetic or endpoint assays. The reagent features a maximum sensitivity of 0.001 EU/mL, the option to use polynomial regression and an economical sample to lysate volume ratio of 4:1. Polynomial regression enables more accurate determination of endotoxin concentrations, especially when a wide range standard curve is used.

The Pyros Kinetix® Flex reader is an optical tube reader that runs both chromogenic and turbidimetric assays. Designed with flexibility and efficiency in mind, the Pyros Kinetix® Flex reader and Pyros® EQS Software combine to provide a complete system that is 21 CFR Part 11 compliant for efficient, accurate endotoxin testing. AJ Meuse, President and CEO of ACC stated “We are very excited about the opportunities that Pyrochrome® and the Pyros Kinetix® Flex reader system is going to offer the market. We understand that quality, regulatory compliance and exceptional technical support are critical components for our customers when deciding who they will trust with their endotoxin release testing and this new system was designed to satisfy our customer’s needs.”

The Pyros Kinetix® Flex reader is available in three configurations: 32, 64 and 96 eight mm wells. Each well is independently timed, allowing the operator to add more samples while a run is in progress. The unit features precise temperature control, solid state design and utilizes reliable Pyroclear® depyrogenated test tubes, which eliminates potential hot wells that are often found when using mircroplates.

ACC is an industry innovator and has been a leading global supplier of endotoxin and glucan detection products and services for nearly four decades. During this time, ACC has supplied our customers with products and services that have helped ensure the safety of their parenteral drugs, biological products and medical devices.

For more information on ACC’s new Chromogenic Endotoxin Detection System incorporating enhanced Pyrochrome® reagent, Pyros® EQS Software and the Pyros Kinetix® Flex Incubating tube reader, contact Associates of Cape Cod, Inc., at 124 Bernard E. Saint Jean Drive, E. Falmouth, MA 02536, 508‐540‐3444, www.acciusa.com.

Cook Pharmica adds E. Morrey Atkinson, Ph.D., to Leadership Team as Vice President of Research and Development, Chief Scientific OfficerE. Morrey Atkinson, Ph.D., has joined Cook Pharmica as vice president of research and development and chief scientific officer, company officials announced recently. Atkinson will be responsible for guiding the scientific direction of Cook Pharmica, relying on his extensive past experience with process development of gene therapies, vaccines, recombinant proteins and monoclonal antibodies.

“Morrey comes to us with a broad background of scientific and business experience in both the domestic and international marketplace,” said Tedd Green, president of Cook Pharmica. “His broad technical, management and leadership experience will be a great strength to our team and we are proud to welcome him to the Cook organization.”

Atkinson has almost two decades of experience in biologics development and has held various leadership positions in biotechnology manufacturing and development. Most recently, he was head of biotechnology manufacturing sciences and technology for Eli Lilly and Company in Kinsale, Ireland.

“I look forward to contributing my experience with biologics development to this organization,” Atkinson said. “With the potential to offer the broadest range of development and production services in the industry, Cook Pharmica has a bright future ahead, and I am excited to join this leadership team.”

Atkinson, who grew up in Indiana and Florida, graduated from Indiana University with a bachelor of science in biology and received his doctorate in biological sciences from Stanford University.

FOSS Releases New Features for VisionFOSS NIRSystems, Inc. has introduced Service Pack 6 for Vision 3.50.

Vision is a software package specifically designed for use with the FOSS NIRSystems Near-Infrared (NIR) laboratory and process analyzers. Vision 3.50 Service Pack 6 is a cumulative service pack and offers support for the latest NIR laboratory and process hardware.

The main new feature added in Service Pack 6:

• The ProFoss instrument has been added to Vision, which includes support for Self-Test, Diagnostics, Data Acquisition, and Routine Analysis. ProFoss is a diode-array instrument and the latest addition to the product line of FOSS NIRSystems.

»


INDuSTRy NEwS »

What are the reasons (medical, social, environmental) in favor of needle Free PFs?Today the statistics show that by far, the most frequent accident with risk of blood contamination in the hospital environment is the needle stick (74%). This is a real critical issue for the health workers (especially the nurses) and the hospital administration that needs to report these accidents and provide insurance coverage for the risks it involves.

Why is it more important for life saving drugs in r2u forma? When time is of essence, a R2U product will limit at the utmost the time it takes to treat a patient, but also it eliminates completely any sources of accidents and medication errors, which happens today still too often.

The French Health authorities reported in 2005 that there could be as many as 190 000 evitable severe adverse effect events each year. These severe events are susceptible to cause death, induce a handicap

AGUETTANT Q&ADanielle LabrecheDirector of Business Development and InnovationLaboratoire AGUETTANT

« IntErvIEW »


or prolong the hospitalization. Data from other countries confirms the size of the issue.

The French Health Authority (AFSSAPS), published in 2009, a report on the Medical Errors, based on 4 years of operations of their “Guichet des Erreurs Médicamenteuses”, an Adverse effect events reporting entity. According to this study, the most common errors are on: drug (42%), dilution (28%); dose (7%), and route of delivery (6%). The source for such errors are attributable to “look alike” packaging (39%), medical procedure errors (27%), use error (12%), and missing information (10%).

If more secure, why are PFs not deployed to all drugs where applicable yet? A lot of critical care drugs have been offered for many years in historical primary packaging: the glass vials or glass ampoule. Years of competition on these products has driven the market to very low prices per unit. Now that the PFS technology exists, but at a higher cost than vial or ampoules, how do we convince the hospital pharmacists to pay more for security? We have to look at time saved for the staff, reduction of medical errors, and let us not under estimate the reduction of wasted drugs and supplies.

Nevertheless, today the PFS market is mostly addressing higher priced drugs, new chemical entities, vaccines and new drug formulation. But some smaller size companies have accepted to live up to the challenge.

What will it take?The industrial supply capacity of pre-fillable syringes is still under the market demand.

The PFS growth expectations are still at double digit growth per year, and so the available capacity focuses on the higher valued markets. But some smaller pharmaceutical companies, such as Aguettant, are looking again at the critical care market, and have achieved to impose their PFS in France on the basis of additional security and less waste. But it took 5 years of R&D investment.

Are there any benefits of plastic over glass? Glass is a fragile material, and heavier than plastic, it offers less ergonomy especially when we are talking of 5 ml, 10ml or 50ml PFS. Also, with plastic, there are more possibilities to propose creative and cost effective designs, for example:

Luer lock connection design, Opening system with tamper evidence and back stopper of the plunger rod.

Types of Errors*

Total In %Error of drug 310 42Error of dilution 205 28Error of dose 54 7Error of route delivery 41 6Other 104 17Total 714 100*AFSSAPS, Medical Errors Reporting Office, June 2009, France

Figure 1. Nature of Exposure

Figure 2. Types of Personnel Concerned

»


InDustry nEWs »

If you are a current user of the FOSS Vision software, please contact us for more information and/or a complete list of new features appearing in version 3.50, Service Pack 6.

The FOSS Vision software is 21 CFR Part 11 compliant and supports PAT through numerous process analysis options and process communication capabilities. The software has an extensive security system with multiple access levels, secure data archiving and report generation as well as database and spreadsheet compatibility. Vision comes with a user-friendly electronic manual with tutorials and data for self taught hands-on method development and software operation.

scilog’s second single-use Pre-Calibrated sensor Patent IssuedSciLog, Inc. announces the issuance of the second of two single-use sensor patents, US 7,788,047 and US 7,857,506 “Disposable, Pre-Calibrated, Pre-Validated Sensors for Use in Bio-Processing Applications.” The first patent was issued in September 2010.

“This patented technology addresses the challenges of the Process Analytical Technology (PAT) initiative as it applies to single-use sensors in downstream bio-processing” commented Karl G. Schick, Ph.D., VP of R&D at SciLog Inc.

SciLog’s single-use, pre-calibrated sensors:

• Provide real-time analytical data.

• Enable pre-sterilized, closed-loop processing environments.

• When used inCan be integrated into single-use, gamma-irradiated fluid pathways (SciLog US patents 6,712,963 and US 7,052,603), enable automation and automated data acquisition for single-use platforms.

SciLog manufactures disposable, pre-calibrated, single-use conductivity (SciCon), pressure (SciPres) and temperature (SciTemp) sensors. , see www.scilog.com www.scilog.com/sensors. The sensors are designed with process scalability in mind. They are available in, five different fluid connection sizes, ranging from Luer (Laboratory) to 1.0” TC (GMP bio-processing), are available in each of the sensor families. Each sensor is NIST traceable by sensor ID and comes with a calibration certificate and appropriate compliance statements. Sensor ID and associated calibration data are stored in a gamma-stable memory residing within the sensor. The stored calibration data is stable for over two years. For more information, go to http://www.scilog.com/sensors.

These patents expand SciLog’s has a significant patent position in downstream, single-use technology. In addition to the newly issued sensor patents, SciLog has received two prior patents (US Patent 6,712,963 and US Patent 7,052,603) that deal with “Single-use Manifolds for Automated, Aseptic Transfer of Solutions in Bio-Processing Applications”. SciLog has also received US Patent 7,410,587 for “Liquid Handling for Filtration and Liquid Chromatography”. Specifically, these patents address the challenges and provide solutions relevant to downstream, single-use purification by tangential flow filtration (TFF), preparative chromatography and normal flow filtration (NFF). In addition, SciLog received US 7,410,587 “Liquid Handling for Filtration and Liquid Chromatography”.

SciLog offers licensing arrangements of its single-use technology to interested parties.

For further information contact:

Juliette Schick,

President, SciLog, Inc.

Ph: 608.824.0500

zymark tPW3 tablet Processing Workstations and APW3 Active Pharmaceutical Ingredient WorkstationThe Zymark TPW3 and APW3 automates sample preparation and analysis for tablets, capsules, blends, creams, lotions, medical devices and suspensions for content uniformity and stability assay testing required in the Pharmaceutical Formulation and Quality Control Labs. Our highly productive workstations not only increase testing throughput and lab productivity, they also improve the quality of analysis by minimizing operator error and eliminating variability compared with labor intensive manual methods. If you are looking for ways to boost your pharmaceutical lab’s productivity through automation, SOTAX has the solution and experience for successful implementation. From method development and validation to standard operation procedures and transfer expertise, SOTAX is your solution for pharmaceutical testing.

SOTAX Corporation

68A Elm Street

Hopkinton, MA 01748 USA

www.sotax.com

Sales Inquiries: 1-888-SOTAXUS

Email Inquiries: [email protected]

Pittcon 2011 Booth #1447

»


« InDustry nEWs

rapid micro Biosystems Partners with life technologies Corporation on Automated microbial Detection– growth Direct™ systemBusinesses team to accelerate microbial detection and identificationRapid Micro Biosystems, a leading provider of automated, non-destructive, rapid microbial detection, announced a sales and marketing agreement with Life Technologies Corporation, a provider of innovative life science solutions. For quality control microbiology customers, the agreement combines best in class automated microbial detection and enumeration with gold-standard microbial identification.

The agreement leverages the strengths of both companies, benefiting customers who struggle daily with time consuming, manual processes, and product safety testing where accuracy and time to results are critical. The goal of the agreement is to maximize industry adoption of the complementary technologies. The Growth Direct™ System from Rapid Micro Biosystems enables rapid microbial detection, and the MicroSEQ® Rapid Microbial Identification System from Life Technologies facilitates accurate bacterial and fungal identification.

“We are excited with the opportunity this alliance presents to our customers,” said Steve Delity, Chief Executive Officer of Rapid Micro Biosystems. “Rapid Micro is the leader in automated rapid detection, and through our collaboration with Life Technologies, the leader in microbial identification, we create a compelling value proposition for the marketplace.”

“The agreement with Rapid Micro Biosystems is part of our larger commitment to address the needs of microbiologists in product quality and safety testing,” said Tony Hunt, General Manager of Pharma Analytics at Life Technologies. “We are pleased to add the Growth Direct™ System for microbial detection to our overall microbiology portfolio.”

For more information, visit www.rapidmicrobio.com/automated_sample_control

AguEttAnt and PFIzEr Enter Into a Patent licensing Agreement for AguEttAnt® self Flushing Infusion Bag AGUETTANT just comes to grant an exclusive licence to PFIZER on its patented self flushing infusion bag, for PFIZER’s systemic antifungal molecules marketed in Europe.

This delivery system, invented and patented in Lyon, France, by AGUETTANT, performs an automatic flushing of the connecting line and perfusion bag after the drug delivery, without the intervention of any medical staff.

This technology offers several benefits for the patient and the medical staff by:

• enabling accurate delivery of the prescribed dose, without the drug loss that is traditionally found trapped in the connecting line;

• reducing the risks of nosocomial infections via reduced manipulations; and eliminating waste linked to the manual preparation of the flushing;

• saving time for the medical staff.

With this innovative delivery system device, both PFIZER and AGUETTANT are striving to improve the safety and quality of care for both patients and healthcare professionals.

nEtzsCh Premier launches line of Deltavita® nanoparticle mills Ideal for Pharmaceutical ApplicationsA full line of mills for laboratory, clinical trial and full-scale productionNETZSCH Premier Technologies, LLC introduced the DeltaVita® line of ultra-fine nanoparticle mills for wet grinding of batches ranging 0.05 to 2000 liters. The new line features NETZSCH’s proprietary ZETA® grinding system and comprises 10 mills, designed to accommodate the entire manufacturing process with repeatability and scalability from testing through development to full-scale production.

Through the use of high-energy, high flow-rate, multiple-pass grinding, NETZSCH’s DeltaVita line achieves excellent repeatability and homogeneous dispersion. In the time it takes an ordinary mill to complete one pass, DeltaVita mills can complete as many as 10 cycles. The ZETA grinding system offers a single-tank process to reduce contamination, maintain precision temperature control and provide an easy-to-clean design.

The DeltaVita 15 through 300 systems use grinding media from 0.05 mm to 0.2 mm for consistent particle reductions to below 200 nanometers. They can be fitted with variable grinding chamber sizes, making them ideal for feasibility studies where the smallest batch sizes are needed to achieve significant test results in a short period of time. Grinding zone parts are manufactured with stabilized Zirconia/Yttrium, a high-strength ceramic, for metal-and contamination-free grinding.

For clinical trial phase production, the DeltaVita 600 can produce batch sizes of one to 6 liters. It features interchangeable agitating systems, optional explosion-proof design and optional PLC control. The DeltaVita 2000 through 60000 provide batch sizes ranging from 50 to 2000 liters.

»


InDustry nEWs »

All sizes of NETZSCH DeltaVita mills feature:

• Wetted parts designed and manufactured according to the latest GMP standards;

• Material, production and calibration certificates included;

• Optional cleaning in place (CIP) and sterilization in place (SIP) capability;

• All indirectly product-wetted surfaces made of stainless steel;

• Optional data recording and formulation management;

• Operator management with password protection for different levels of security;

• Various materials (such as ZrO2, stainless steel 316 or nylon) available for custom chamber design;

• Splash-proof machine stand;

• Comprehensive testing and qualification documentation; and

• Training sessions and seminars available.

All NETZSCH pharmaceutical process equipment, including the DeltaVita series, meets the high demands of the pharmaceutical industry. NETZSCH has a dedicated team for pharmaceutical applications experienced in the industry-specific requirements for testing, validation, and documentation including FAT, IQ, OQ, URS, FRA and DDS.

For more information, visit http://www.netzsch-grinding.com or call 484-879-2020.

Crystal PharmatechCrystal Pharmatech Co., Ltd is currently the only high-quality, dedicated solid state research company based in China. Our researchers have extensive experience in top pharmaceutical companies with API and drug product development. Whether you are a top-tier innovator company or a pre-emerging biotech, Crystal Pharmatech can handle all of your solid-state research and development needs. We offer a full range of services from being the solid state characterization arm of your company to consultation or training on a specific issue.

“Solid state research and development support is critical to speeding up drug development, lowering costs, and providing the highest quality product to innovator pharmaceutical companies.” explains Alex M. Chen, CEO of Crystal Pharmatech. “China is currently the top choice for pharmaceutical outsourcing owing to the strong support from government and vast talent pool of relatively low cost labor. It is inevitable that China-based companies focused on providing solid state R&D solutions will be an integral component to the outsourcing package.”

Besides providing contracted research services, Crystal Pharmatech Co., Ltd, will also be developing new technologies and proprietary devices that will improve efficiency and drive down development costs for all pharmaceutical companies.

For more information, visit: http://www.crystalpharmatech.com/

Particle sizing systemsWhen Size Really Matters, You can Count on Particle Sizing SystemsParticle Sizing Systems announces that it is updating its look with a new logo. Our company has evolved over the past 33 years as a major force in the particle sizing industry. We have had a few logos along the way. However, the one thing that remains constant with PSS is that we continue to patent and manufacture particle sizing instrumentation that sets us apart from the others that are available in the market place today. We have dedicated and continue to dedicate ourselves to developing leading edge technology and producing innovative instruments that offer unique, powerful capabilities for laboratory and production environments. We will continue working to serve, support and provide new innovations in the field of particle sizing.

Please visit our new website to see all of the particle sizing instruments that we have to offer.

www.pssnicomp.com

B&W tek, Inc Announces Improved Quest™ series miniature Fiber optic spectrometersB&W Tek, Inc., an advanced instrumentation company producing optical spectroscopy and laser systems announces the updated Quest™ series of miniature fiber optic spectrometers. The Quest™ series of high performance miniature fiber optic spectrometers now features an ultralow thermal drift spectrum of ~19 counts/C typical and a faster readout speed of >2.0MHz. As an added bonus, the Quest™ series is now available with optional RS232 communication interface for convenient integration into larger systems.

“The fast read out speed of the new Quest™ series spectrometers makes them ideal for high speed applications such as LED binning and sorting,” says Robert Chimenti, Marketing Manager for B&W Tek, Inc. “We feel that this will strengthen our position as a leader in the LED characterization market.”

The Quest™ series employs both a traditional crossed Czerny Turner spectrograph (Quest™ X) as well as an unfolded Czerny Turner spectrograph (Quest™ U) which minimizes stray light in the UV region. The series is equipped with 2048 elements linear CCD array, built-in 16-bit digitizer, and an externally synchronized trigger.

For more information on the Quest™ series, please visit http://www.bwtek.com/product/spectrometer/.

« ClAssIFIED ADvErtIsEmEnts »


»

www.acciusa.com 800.LAL.TEST (525.8378)[email protected]

If You Don’t, You’re Probably Paying Too Much.If You Don’t,If You Don’t,

DO YOU KNOW YOUR ENDOTOXIN COST PER TEST?

Contact us by calling 800-724-4158www.advancedscientifics.com

single use systems

Achieving ‘Faster Time to First in Man’Capsugel R&D benchtop machines.

www.capsugel.com

APRBnr_17373_v1_2-19-09.indd 1 2/19/09 4:31:14 PMInnovative solutions.Global capabilities.For more information, contact us at +1 866 720 3148, email us at [email protected], or visit www.catalent.com

©2008 Catalent Pharma Solutions

PAT-aligned endotoxin testingwww.laboftomorrow.com

Rapid Testing Solutionswww.lonza.com/rts

Get results now

RTS_classified.indd 1 8/24/10 3:42 PM

C

M

Y

CM

MY

CY

CMY

K

11-4237_KSDbiomall82.5x25.7_DM.pdf 24/01/11 13:52:59

Call +1 866-PATHEON (+1 866-728-4366) or email [email protected]

Why limit your compound to just one bioavailability enhancement technology?

Configurable solutions. Flexible, ready-to-connect unit operations.

www.sartorius-stedim.com/flexactturning science into solutions

Configurable solutions. Flexible, ready-to-connect unit operations.

www.sartorius-stedim.com/flexactturning science into solutions

»


ADvErtIsEr’s InDEX »

Company Page # Phone # Web Address

AAPS IBC 703-243-2800 www.aapspharmaceutica.com

AB Sciex 19 877-740-2129 www.absciex.com

Accugenix, Inc. 54 302-292-8888 www.accugenix.com

Advanced Scientifics, Inc. 63 [email protected] www.advancedscientifics.com

Aguettant 77 +33 (0)4 78 61 51 41 www.aguettant.com

Alfa Aesar 85 978-521-6300 www.alfa.com

Associates of Cape Cod 6,53 888-395-2221 www.acciusa.com

Blake Hotel 80 888-664-6835 blakehotelnc.com

Bruker 25 888-4BRUKER www.bruker.com

Cambridge Healthtech Institute 65 888-999-6288 www.healthtech.com

Capsugel 1 888-783-6361 www.capsugel.com

Catalent Pharma Solutions 67 866-720-3148 www.catalent.com

Charles River IFC [email protected] www.criver.com

Crystal Pharmatech 69 855-546-4986 www.crystalpharmatech.com

Dionex 15 408-737-0700 www.dionex.com/chromeleon7

Distek, Inc. 45 888-234-7835 www.distekinc.com

Enwave Optronics 40 949-955-0258 www.enwaveopt.com

GE Healthcare Life Sciences 60 800-526-3593 www.gelifesciences.com

Horiba Scientific 41 732-494-8660 www.horiba.com/scientific

ICDD - International Centre for Diffraction Data 27 610-325-9814 www.icdd.com

Intercontinental Hotel 57 800.424.6835 www.intercontinental.com/baltimore

Interphex 11 203.840.5447 www.interphex.com

Kaiser Optical Systems, Inc. 37 734-665-8083 www.kosi.com

Life Technologies 64 760-603-7200 www.lifetechnologies.com

Lonza 51 800-638-8174 www.lonza.com/moda

Meggle Group 83 914-682-6891 www.meggle-pharma.com

Mettler Toledo, Inc. 43 800-METTLER www.mt.com

Netzsch 31 484-879-2020 www.netzsch-grinding.com/pharma

Pall Life Sciences 59 [email protected] www.pall.com/biopharm

PANalytical 23 +31 (0)546 534444 www.panalytical.com

Particle Sizing Systems 33 727-846-0866 www.pssnicomp.com

Patheon 71 919-226-3200 www.patheon.com

Parenteral Drug Association (PDA) 3 301-656-5900 www.pda.org

Perfex 5 800-848-8483 www.perfexonline.com

Phenomenex Inc. 13 310-212-0555 & [email protected] www.phenomenex.com

Rapid Micro BioSystems 55 781-271-1444 www.rapidmicrobio.com

Sartorius Stedim North America 9 800-368-7178 www.sartorius-stedim.com

Sotax 47 1-888-SOTAXUS www.sotax.com

Thermo Fisher Scientific OBC 978-642-1132 www.thermo.com

ToxExpo 29 703-438-3115 www.toxexpo.com

Waters Corporation 17 800-252-4752 www.waters.com

Watson-Marlow Pumps Group 61 800-282-8823 www.wmpg.com

West Pharmaceuticals 75 800-345-9800 www.westpharma.com

Witec 39 865-984-4445 www.witec.de

»

The contact directory is for information purposes only. While every effort has been made to ensure it is accurate and complete, any errors or omissions are not the responsibility of the publisher.

Mark Your Calendar!

For Up-To-Date Informationwww.aapspharmaceutica.com/nationalbiotech

MAY 16~182011Hilton San Francisco

Union Square

SAN FRANCISCO CA

C

M

Y

CM

MY

CY

CMY

K

©20

11 T

herm

o Fi

sher

Sci

enti

fic In

c. A

ll ri

ghts

res

erve

d.

Raw material identification just got exciting!

Introducing an exciting new advance in raw material

identification – the Thermo Scientific TruScan RM analyzer.

Our lightest, fastest and most portable handheld analyzer

utilizes the power of Raman spectroscopy to increase incoming

inspection rates – saving you time and money.

Combining a state-of-the-art optical platform with enhanced

decision algorithms, the analyzer simplifies everything from

method development to routine authentication.

The TruScan® RM analyzer lets you perform identification of a

broad range of chemical compounds through sealed packaging

with more speed and reliability than ever before.

Work faster. Work smarter. Get excited!

Learn more at www.thermoscientific.com/RMID

Moving science forward

Thermo Scientific TruScan RM AnalyzerProviding GMP facilities with cost-effective and efficient raw material identification.

russell_apr_20110102

Documents

Transcript of russell_apr_20110102