Impurity Profiles and Degradation Products in Raw …...Some of the biggest culprits: the quality of...
Transcript of Impurity Profiles and Degradation Products in Raw …...Some of the biggest culprits: the quality of...
©2013 Waters Corporation 1
Impurity Profiles and Degradation Products in Raw Materials and Pharmaceutical Products
Michael D. Jones
Ignatius (Iggy) Kass, Ph.D.
Sr. Field Marketing Manager
www.waters.com
Confidential
©2013 Waters Corporation 2
Content
Understanding your Challenges:
– Risks of impurities in pharmaceuticals
Strategies for Impurity Profiling
– Quality Control, batch acceptance
– “Impurity Profiling”
Application examples
Summary
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The risk of impurities
Unwanted chemicals influence the efficacy and safety of the pharmaceutical products.1
– “Any component of the drug product that is not the chemical entity defined as the drug substance or an excipient in the drug product.”2 (ICH Q6A: Specifications).
– An excipient is a pharmacologically inactive substance formulated with the active ingredient ("API") of a medication
Impurity profiling (i.e., the identity as well as the quantity of impurity in the pharmaceuticals) is receiving important critical attention from regulatory authorities.1
Various regulatory authorities like ICH (International Conference on Harmonization), USFDA, Canadian Drug and Health Agency are emphasizing on the purity requirements and the identification of impurities in Active Pharmaceutical Ingredient’s (API’s).1
1. Tegeli et.al, IJDFR volume 2 Issue 4, Jul-Aug.2011
2. ICH Q6A: Specifications
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Regulations to promote good science Impurity Thresholds for Drug Substances
Detect
– Know that it is there
Identify
– Know what it is
Qualify
– Know that it is safe
Maximum Daily Dose
Threshold
Reporting Identification Qualification
≤ 2 g/day 0.05%
0.10% or
1.0 mg/day intake (lower)
0.15% or
1.0 mg/day intake (lower)
> 2 g/day 0.03% 0.05% 0.05%
Guidance for Industry, Q3A Impurities in New Drug Substances, February 2003; \\CDS029\CDERGUID\4164fnl.doc Draft Guidance for Industry, ANDAs: Impurities in Drug Substances, January 2005; J:!GUIDANC\6422dft.doc
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Impurity Profiling Business Impact on Operations
Acceptance or rejection of raw material for production
– Impurity profiles define material acceptance or rejection
– Out of specification (OOS) : Reject raw materials
Sign-off of finished API or drug product for release
– Ensures quality of the product
– Impacts time to market and customer satisfaction
Adhere to cGMPs and CMC regulatory requirements
– Regulatory filings dependent on time to complete studies
– Results must be accessible and confidently defendable to regulatory
authorities
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Analytical Challenges
Throughput – Multiple batches are required to be analyzed
– Shorter turn round times to increase profitability
Sensitivity – Low level impurities can pose a serious issues with genotoxicity
– Sample impurity ”finger print’ used to support counterfeit activities
Specificity – Ability to quickly detect drug or excipient related material simplifies
impurity detection and identification
Ease of collecting required data – Transfer procedures within and between labs
– Employee turnover
Informatics – Getting results that drive decisions
– in silico tools (modeling, prediction) utilize prior knowledge to allow analytst to focus on the more challenging tasks
– Merging data from different sources (experiments, labs, geographies, etc)
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Potential Impurities
Degradants (Forced degradation studies)
– Acid, base, heat, light, etc.
Synthetic intermediates
In process 2007 Viracept recall, Roche Pharmaceuticals, ethyl mesylate, genotoxic. - Tracked
down to a reaction between the active ingredient and chemicals used in Cleaning in
Manufacturing.
Genotoxins
And anything else……
“Alerting functional moiety” that is shared with the impurity or API structure
- At any level
Muller, L., Mauthe, R.J., 2006. A rationale for determining, testing and controlling specific impurities in pharmaceuticals that possess
potential for genotoxicity. Regulatory Toxicology and Pharmacology 44, 198-211
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Sensitivity Required for Genotoxins
Impurities should be reported at levels in excess of the 0.03 – 0.05% (300 – 500 ppm, by weight)
range according to ICH_Q3A_R2 guidelines. Genotoxic impurities, or potentially genotoxic impurities,
must be controlled at levels significantly lower than this. Typically, developing limit tests of less than
50 ppm for highly toxic impurities is readily achievable. However it can be difficult to develop a test
to control a particular genotoxic impurity at the current EMEA guideline limit of 1.5 ppm (0.00015%
w/w) or less.
– AAL Website
"For genotoxic impurities we need very sensitive and selective methods. One needs higher
sensitivity to determine ppm-level impurities and selective methods to separate low levels of
genotoxic impurities from base line noise and other organic impurities. The typical HPLC methods with
a nonspecific detector (e.g., UV) that are used to measure organic impurities may not be appropriate
to quantitate low ppm levels of genotoxic impurities.“
– Maribel Rios, Pharmaceutical Technology
– 10 – 100 times more sensitivity needed
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The safety and business risk even if you meet regulations
Some of the biggest culprits: the quality of raw materials, faulty labeling
and packaging and contamination.1
A number of factors are fueling the recall surge. The stampede by
drugmakers to be first to bring generic versions to market, after drugs lose
patent protection, is one of them.1
– "The first applicant typically gets the lion's share of the business for the new drug,"
said Cox.
– In their hurry, drugmakers sometimes fail to spend enough time learning how best
to make the drug.1
2011 J&J Recalls helped push net income down 12 percent ($900 million) 2
1 http://money.cnn.com/2010/08/16/news/companies/drug_recall_surge/index.htm 2 http://www.bloomberg.com/news/2011-01-25/j-j-profit-declines-12-on-product-recalls-2011-outlook-trails-forecast.html 3Tablets & Capsules 2012
1 3
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Strategies
I. Quality Control
– Impurities are well defined: USP methods and standards
II. Impurity Profiling
– Full characterization: Define what may be present
– Where to begin?
• Synthetic routes – In process impurities
• Degradation – Heat, Light, Acid/base...
• Previous production on manufacturing line
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I. QC testing WATERS UPLC SOLUTION
Quick, easy and cost effective
– Typically LC/UV
– Impurity profile is already known at this point
o Information you have is the compounds retention time..
HPLC
HPLC
UPLC
UPLC
Speed
Resolution
Sensitivity
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QC testing
Fast, easy and cost effective
– Typically LC/UV
– Impurity profile is already know at this point
o Information you have is the compounds retention time..
Example
USP method and known impurities
API Related
Known Impurities
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QC testing
Fast, easy and cost effective
– Typically LC/UV
– Impurity profile is already know at this point
o Information you have is the compounds retention time..
Example
USP method and known impurities
API
Known Impurities
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QC testing
Fast, easy and cost effective
– Typically LC/UV
– Impurity profile is already know at this point
o Information you have is the compounds retention time..
Example
USP method and known impurities
API
Known Impurities
What is it?
?
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©2013 Waters Corporation 15 Michael D. Jones, PBO
Time
%
Potential Risks
API
Known Impurities
?
What if we look at these peaks by Mass Spectrometry?
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©2013 Waters Corporation 16 Michael D. Jones, PBO
1: MS gives an extra dimension of separation:
210 350nm
100
%
210 350nm
100
%
210 350nm
100
%
200 300 400m/z
100
%309.1
311.1
200 300 400m/z
100
%
309.1
311.1287.1
200 300 400m/z
100
% 309.1
287.1
Mass Spectrometry (Single Quadrupole)
Time
%
API
Known Impurities
?
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What does Contamination look like? Supplier Comparisons: Budesonide
Supplier A
Supplier B
Supplier C-Lot1
Supplier C-Lot2
Representative MS TIC Chromatograms
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II. Waters Solution for Complete Impurity Profiling
UPLC – Speed of analysis
– Challenging separations of related compounds
o Peak Capacity
Dynamic Range – Sensitivity for low level peaks
– Mass Measurement Accuracy for High Intensity peaks
Informatics – Simplicity of processing / Setup
– Easy operation for known analytes
– Detection of unknowns o Automatic calculation of elemental composition
o Confirmation / Elucidation with MSE data sets
– Correlation of PDA Data in reporting
Detect
– know that it is there
Identify
– Know what it is
Qualify
– Know that it is safe
– (Biology and safety
studies)
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Summary of Major Simvastatin Degradation Products*
MSUVMSUVMSUVMSUVMSUV
3.970
4.423
5.209
5.254
5.699
6.222
6.322
6.881
7.257
3.862
3.841
3.594
3.449
3.137
3.107
2.986
2.938
2.397
2.308
2.191
2.161
2.116
2.062
1.926
1.886
1.760
1.699
Photo DegradationPeroxide
Oxidation
Temperature
Degradation
Base HydrolysisAcid Hydrolysis
Peak
Rt (min)MSUVMSUVMSUVMSUVMSUV
3.970
4.423
5.209
5.254
5.699
6.222
6.322
6.881
7.257
3.862
3.841
3.594
3.449
3.137
3.107
2.986
2.938
2.397
2.308
2.191
2.161
2.116
2.062
1.926
1.886
1.760
1.699
Photo DegradationPeroxide
Oxidation
Temperature
Degradation
Base HydrolysisAcid Hydrolysis
Peak
Rt (min)
*Does not include impurities detected in Simvastatin standard.
MS Detection is VERY Sensitive, But still a Complementary Technique
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Typical Workflow
Flag questionable impurities
Isolate impurities by API removal or impurity enrichment
Perform MS/MS or MSn structural studies
Confirm by NMR
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Design a Waters Strategy
Separate A well developed impurity
profile method is KEY
Identify MSE
Metabolynx XS
Elucidate MassFragment
Isolate Mass Directed Autopurification
Scalable Chemistries
?
C4H7N2O3
NN
S
N
O
OH
Man
ag
e
Compile
Track
File
NuGenesis SDMS
NuGenesis SDMS
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Method Development
Selectivity [α]
Stationary Phase
Organic Modifier
Mobile Phase pH
Column Manager
•Stack to support up to 6 columns
•Independent heating/cooling zones
Sample Manager—FTN
•Direct injection simplifies mode of injection selection
•Injection volume flexibility without reconfiguring injector
Quaternary Solvent Manager with Optional Solvent Select Valve
•Up to 9 solvents with integrated solvent selection valve on the D line (1-6 solvents)
•Select study variables
•Define study ranges Experiment
Design
•Conduct formal experimental design
•Analyze results, build equations of variable effects
Knowledge Space
•Define best conditions
•Define robustness
Design Space
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Impurity ID/Elucidation
Generic methodologies still provide maximum coverage Automated data interpretation Identify drug related compounds Elemental Composition /Structural elucidation
The most sensitive, exact-mass,
quantitative and qualitative benchtop
MS system
Uses UPLC/MSE, a simple, patented
method of data acquisition to
comprehensively catalog complex
samples in a single analysis
Provides the most complete system
solutions backed by superior support to
ensure customer success
Guarantees maximum system
performance and usability through our
implementation of Engineered
SimplicityTM
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Identification …detection in complex mixtures
Intensity 6.04 x 106
Intensity 535
Mass Measurement Accuracy and Quantitative Dynamic Range Concentration of drug very high Excipients add to complexity (PEG, etc.)
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Alternate Scanning LC-MS (LC-MSE) …time resolved mass measurements Parallel LCMS (LCMSE)
…a dual function LC-MS protocol
MSE
is a UNBIASED process
MSE
is a CONTINUOUS process
1 sec
LE
EE
LE
EE
EE EE
D.
m
All ions all the time
Very Simple Setup
Bateman et al, Anal Chem (2002) Silva et al, MCP (2005) *Patented technology
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Identification with MSMS Example: Nefafzodone
N
N
N
O
CH3
ON
N
Cl
N
N
N
O
CH3
ON
N
Cl
NCH3
N
N
O
CH3
N
N
Cl
N
N
N
O
CH3
OCH3
CH3N
N
Cl
NH
N
N
O
CH3
CH3
MS (Detect)
MSMS (Know what it is) Confidential
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Systematic Bond Disconnection and Exact Mass
MassFragment Software does this automatically
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Parent drug fragment ion characterization with MassFragment
Nefazodone
Nefazodone fragments
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Impurity Isolation for alternate techniques (NMR, Optical, etc..)
Mass directed purification offers
far greater specificity and
efficiency than traditional UV
directed systems
With both analytical and
preparative performance available
as standard, method
development, purification and
fraction analysis are performed
automatically, and without
compromise
OpenAccess software provides
access to the advanced fraction
collection functionality by even
the most novice users
Crude mixture
? LC/MS
purification
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Key Contributors
Michael D Jones
Warren Potts
Marian Twohig
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©2013 Waters Corporation 31 Michael D. Jones, PBO
Test Case Compound
Quetiapine fumarate (Anti-Depressant)
Chemical Name:- Bis[2-(2-[4-(dibenzo[b,f][1,4]-thiazepin-11-
yl)piperazin-1-yl] ethoxy) ethanol] fumarate
Quetiapine fumarate has no chiral centers and only one morphological
entity has been detected throughout development
N
N
S
N O
OH
Quetiapine
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Test Case 1: Qualitative Profiling Management Confirm Known Impurities
A Workflow Approach for the Identification and
Structural Elucidation of Impurities of Quetiapine
Hemifumarate Drug Substance
Increase Visualization using Metabolynx w/ MassFragment
Search for API Relationships
Analyze using MSE
Fragment Ion Analysis
*Optional
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Quetiapine Impurity Profile
Michael D. Jones
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MetaboLynx XS Browser
Impurity Results Table
Corresponding impurity spectra
Combined impurity trace
Extracted impurity ion trace
Control v. Analyte Known v. Unknowns
Chromatography data
Exact Mass data Apply Mass
Defect Filters Central
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Rapidly Lists Detected Peaks:
Automatically Generates a List of Impurities within Minutes Confidential
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Quetiapine Impurity Profile has many UNKNOWN Peaks
Michael D. Jones
Quetiapine Confidential
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One Quetiapine Impurity MSE Result
Michael D. Jones
Com
mon F
ragm
ent
Ions t
o A
PI
• 279.1030
• 253.1522
• 221.1132
• 210.0520
• 183.0358
• 139.0473 MSE spectra of Quetiapine Impurity [M+H] = 340.1536
Use MSE data to identify structural components you may already know!
N
N
S
N O
OH
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MassFragment Impurity Investigation of m/z 456.2305
OH
CH3
CH3
N
N
S
N
O
O
MassFragment summary report
Michael D. Jones
MSMS spectrum
MOL file used for [M+H] = 456.2305
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FRAGMENT ANALYSIS
Michael D. Jones
Data Mines Known v. Unknowns
Precursor & Product Ions
Review XIC in Browser
Links to MassFragment
High - CE scan window
Expected/unexpected impurities
Chromatographic common precursors of selected ion
Rapidly Identify Related Impurities
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Test Case2 : Targeted Identification and Isolation of a Previously Unidentified Impurity
Identification and Characterization of an Isolated
Impurity Fraction: Analysis of Impurity 402 found in
Quetiapine Fumarate
Elucidate with MassFragment
Generate Elecomp and proposed structure
Analyze Fraction by MSE
Scale Methodology for Preparative Isolation
Identify Unknown
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UPLC Impurity Profile Method: Acid Degradation of Quetiapine
Quetiapine
Major Unknown
impurity LC System: Waters® ACQUITY UPLC® System
Column: ACQUITY UPLC BEH C18 Column
2.1 x 100 mm, 1.7 µm
Column Temp: 65 ˚C
Flow Rate: 800 µL/min.
Mobile Phase A: 10mM Ammonium Bicarbonate,
pH 9.0
Mobile Phase B: Acetonitrile
Gradient: 15-39%B for 10.5min,
39-43%B to 14.4minutes,
43-95%B to 18min,
hold at 95% for a total run time of 20min
1.326 Peak 1 - SQ 1: MS Scan 1: 50.00-600.00 ES+, Centroid, CV=Tune
402.1
Inte
nsit
y
0.0
4.0x106
8.0x106
1.2x107
1.6x107
m/z110.00 220.00 330.00 440.00 550.00
m/z=402
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Transfer from UPLC to Preparatory
UPLC Column 2.1 x 50 mm,
1.7µm
Time (min)
Flow Rate (mL/min)
%A %B Segment Duration
Time (min)
Segment Duration
(c.v)
0.0 0.8 95 5 0 0
2.0 0.8 40 60 2 9.25
3.0 0.8 5 95 1 4.62
Prep Column 19 x 100 mm,
5 µm
Time (min)
Flow Rate (mL/min)
%A %B Segment Duration
Time (min)
Segment Duration
(c.v)
0.0 25 95 5 0 0
10.5 25 40 60 10.5 9.25
15.7 25 5 95 5.2 4.62
Calculated Migration to Prep from UPLC
Diehl, Diane M. et al. Method Migration From UPLC Technology to Preparative HPLC, Waters App Note 720002375EN
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Mass Directed Auto-Purification
basic conditions
Time6.00 7.00 8.00 9.00 10.00 11.00
%
0
100
Que_Aq_Only_Acid_Deg_22 1: Scan ES+ TIC
2.40e8
Pump 2545 Binary Gradient Module
Inject/Collect 2767 Sample Manager
Detector 2998 Photodiode Array
Mass Spec 3100 Mass Detector
Column 19 X 100mm XBridge, 5 µm
Solvent A 10 mm Ammonium
Bicarbonate, pH 9
Solvent B Acetonitrile
Flow Rate 25 mL/min
Gradient 5% - 60%B over 10.5 min.
flushed for approx 5 minutes
95% organic
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Verification of Pooled m/z 402 Fractions
1.3
25
Inten
sity
0.0
6.0x106
1.2x107
1.8x107
2.4x107
Minutes0.60 1.20 1.80 2.40 3.00
AU
0.000
0.003
0.006
0.009
0.012
Minutes0.00 0.60 1.20 1.80 2.40 3.00
1.309 Peak 1
302.3
379.2 396.0
AU
0.000
0.010
0.020
0.030
0.040
nm240.00 280.00 320.00 360.00
ES+ TIC UV 250nm
1.326 Peak 1 - SQ 1: MS Scan 1: 50.00-600.00 ES+, Centroid, CV=Tune
402.1
Inte
nsit
y
0.0
4.0x106
8.0x106
1.2x107
1.6x107
m/z110.00 220.00 330.00 440.00 550.00
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Elemental Composition
Molecular
Formulas
Mass Error
Values
i-Fit
Settings
Flexibility
Elecomp
Provides
Elemental Composition report for quetiapine impurity with an observed accurate mass of m/z 402.1838.
= Quetiapine + H2O
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Possible Structures
O
NNH
S
N
O
OH
O
N
NO
OH
N
S
O
N
NH2
S N
O
OH
S-oxide and C=N reduction based structure?
C21 H28 N3 O3 S
C=N cleavage creating a Nitroso based structure?
C21 H28 N3 O3 S
C=N cleavage creating an Amino/ketone based
structure?
C21 H28 N3 O3 S
S
N
N
N
O
OH
?
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Ruling out Proposed Structure “B” Nitroso- based compound
O
N
NO
OH
N
S
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Ruling Out Proposed Structure “A” S-oxide- based compound
O
NNH
S
N
O
OHConfidential
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MassFragment
O NNH2
S
N
O
OH
[M+H] = 402
Report based on:
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Accurate Mass MS/MS
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13 C NMR (From Collected Fractions)
O NNH2
S
N
O
OHConfidential
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H-NMR (From Collected Fractions)
O NNH2
S
N
O
OH
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Conclusions
Chromatographic separation and retention time reproducibility is critical to imparting confidence in the quality of your products. Some laboratories are implementing simple ms detection to minimize risk
Accurate Mass - Mass Spectrometry adds another dimension of identification for routine screening and plays a critical role in defining impurity profiles
Often scale-up is required to utilize less sensitive confirmatory analysis
Informatics that incorporate chemical know-how increase productivity by providing a route to fast and accurate reporting
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Key advantages of Waters Systems
Comprehensively catalogue complex samples in a single
analysis with consistency, specificity and wide dynamic range
with innovative UPLC and MSE technology
Engineered simplicity and IntelliStart makes exact mass
routine.
Informatics offering an automated, structure driven, chemically
intelligent workflow for complete data set reporting
Providing services bridging workflows across organizations
throughout from preclinical to final marketing
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?
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References
SIGNIFICANCE OF IMPURITY PROFILING: A REVIEW
V.S. Tegeli*, G.B. Gajeli, G.K. Chougule, Y.S. Thorat, U.S. Shivsharan, , S.T. Kumbhar
, DSTSM’s College of Pharmacy, Solapur., Maharashtra, India. 413003
– Tegeli et.al, IJDFR volume 2 Issue 4, Jul-Aug.2011
Recent trends in the impurity profile of pharmaceuticals
– Kavita Pilaniya, Harish K. Chandrawanshi,1 Urmila Pilaniya,1 Pooja Manchandani,2 Pratishtha Jain,3 and
Nitin Singh3
– J Adv Pharm Technol Res. 2010 Jul-Sep; 1(3): 302–310.
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