Presentation Title Arial 28pt Bold Agilent Blue · 1 2009 - Stimuli article „Transfer of...

July 22, 2011Confidentiality Label

1

Infinitely betterinstrument to instrument method transfer

Agilent 1290 Infinity withIntelligent System Emulation Technology (ISET)

Christian GotenfelsProduct Manager

Agenda

• Why is instrument to instrument method transfer soimportant?

• Parameter affecting instrument to instrument method transfer

• What is ISET?

• Performance examples

• Comparison with current solutions

• Summary

Why is instrument to instrument method transfer so important? - Historical view

July 22, 2011

2004 - 483 incidence observed by the FDA 1

2009 - Stimuli article „Transfer of Analytical Procedure:A Proposal for a New General Information Chapter 2

2011 - Proposal for a New General USP chapter <1224>

1 Johnson Y, „An FDA perspective on method transfer on chromatographic analytical methods in the pharmaceutical industry,“ Chrom. Forum Delaware, October 2004

2 Oscar Quattrocchi, Greg Martin Dennis Runser, Robert Iser, Fred Xi, Horacio Pappac, “Transfer of Analytical Procedures: A Proposal for a New General Information Chapter,” Pharmacopeia Forum, Vol. 35(5) [Sept.- Oct.2009]

Method Transfer Workflow

Location A Regulated environment

Discovery Quality Control

Location B Regulated environment

Discovery Development Quality Control

Development



Discovery Quality Control

CRO

Development

Development



Discovery Quality ControlDevelopment

CMO Regulated environment

Quality Control


Methods from Literature: Scientific journals, applicationnotes, etc

Instrumentation:1100 Series, 1200 Series, Alliance, Acquity, etc

?

Design Differences (U)HPLC Systems

• Delay volumes(gradient formation)

• Power range(flow x pressure)

• Extra column volume

• Temperature

• Data rates

• Sensitivity

Problem:Instrument to Instrument Method Transferability• Not possible• Requires Re-development• Requires Re- validation

Resulting in huge additional:cost factor:

Instrument to Instrument Method Transferability- Important Parameters

Pump

Sampler

ColumnThermostat

Detector

Del

ay v

olum

eE

xtra

col

umn

volu

me

Retention TimeResolution

Retention TimeResolutionSensitivity

Retention TimeResolution

ResolutionRetention TimeSensitivity

Delay volumeGradient mixing behavior Pressure x flow rate

Delay volumeExtra column volumeInjection volume

Temperature profileExtra column volume

Data rateExtra column volumePath-length

Method Transferability: 1260 Quat / 1220 GradFull HPLC Methods Compatibility – Riskless Replacement

1100 / 1200 Series Quaternary system

Column: 4.6 x 150 mm, 5 µmGradient: 0 min 20 % B

10 min 95 % Flow rate: 1 mL/min

min0 2 4 6 8 10

Run existing conventional HPLC methods and get same result !

1260 / 1220 Infinity LC Quat / Gradient system

1100/1200 Series HPLC

1260 InfinityQuaternary LC

1200 Series RRLC – Std.

1260 InfinityBinary LC

Max Flow Rate 5 mL/min 5 mL/min 5mL/min 5 mL/minDelay Volume 900-1200 µL 900-1200 µL 900-1200 µL* 900-1200 µL*Capillary ID 0.17mm 0.17mm 0.17mm 0.17mmDisp. Vol. w/o cell 15µl 15µl 15µl 15µlInjection Principle Variable Loop Variable Loop Variable Loop Variable LoopInj.Volume – Std/Ext. 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µLArea RSD <0.25 % <0.25 % <0.25 % <0.25 %Oven Design A A A AColumn Length 300 mm 300 mm 300 mm 300 mm

* Smaller delay volumes possible

Seamless Method Transfer 1260/1220 InfinityBy unchanged critical specification

Optimized for 3 – 4.6mm ID

Columns

1100/1200 Series HPLC

1260 InfinityQuaternary LC

1200 Series RRLC – Std.




Max Flow Rate 5 mL/min 5 mL/min 5mL/min 5 mL/min 5 mL/min 5 mL/minDelay Volume 900-1200 µL 900-1200 µL 900-1200 µL* 900-1200 µL* 340 µL** 10 – 110 µL*Capillary ID 0.17mm 0.17mm 0.17mm 0.17mm 0.12mm** 0.12mm*Disp. Vol. w/o cell 15µl 15µl 15µl 15µl 7.5µL** 7.5µL*Injection Principle Variable Loop Variable Loop Variable Loop Variable Loop Variable Loop Variable LoopInj.Vol. – Std/Ext. 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 µLArea RSD <0.25 % <0.25 % <0.25 % <0.25 % <0.25 % <0.25 %Oven Design A A A A A AColumn Length 300 mm 300 mm 300 mm 300 mm 300 mm 300 mm

* Smaller delay volumes possible

Seamless Method Transfer What if critical specification change ...? * Optimized for 2.1 - 4.6mm ID

** Optimized for 2.1mm ID

July 22, 2011

Gradient Analyses with Different LC Systems- Impact of delay volume and mixing behavior

2 4 6 8 10 12

mAU

0

50

100

150

200

2

.74

7

3.0

49 8

.12

1 8

.32

3

8.7

47

8.9

30

9.5

45 9

.97

5

12

.00

5 1

2.2

15

12

.39

3

2 4 6 8 10 12

mAU

0

50

100

150

200

DAD1 C, DAD1B, DAD: Signal B, 275 nm/Bw:4 nm Ref 400 nm/Bw:60 nm (B:\RIC\AGI...A\GV 1PHARMA\HPLC000001.D)

2.7

44 2

.88

4 6.0

88

6.3

28

6.6

73

6.8

72

7.4

80

7.9

23

9.9

37

10

.24

9

1200 Series LC Quatenary Pump

1290 Infinity LC

Sample: 0.5% impurities in formulation (metoclopramide)Xbridge C18, 150x3 mm, 3.5 µm dp, 0.45 ml/min, Eluent: A =0 .25 % AmAc, B = ACN, Gradient: 0-15 min; 5-57 % B

The result:• Difference in RT and Resolution• One peak is missing!

Instrument to Instrument Method Transferability- Impact of delay volume and mixing behavior

What happens with a programmed gradient?

time

90%

10%

response

Programmed gradient step

1200 Series Binary Pump SL

1290 Infinity Binary Pump

1200 Series

1290 Infinity

Total delay volume of the system(sum of capillaries, mixer, cells, valves..)

Delayvolume

Gradient slope/Mixing behavior

July 22, 2011

Intelligent System Emulation Technology

(ISET)The Solution

NEW

What is ISET?- Two components

A. Best in class performance of the 1290 Infinity LCBroad power range, ultra-low delay volume, superior sensitivity, unmatched flow and composition accuracy

What is ISET?- Two components

A. Best in class performance of the 1290 Infinity LCBroad power range, ultra-low delay volume, superior sensitivity, unmatched flow and composition accuracy

B. The revolutionary ISET emulationalgorithmFull characterization of target LC systemsCreation of emulation function

FA

Peak 3Overlay of 10 runs

RSD = 0.005 %

Method Transferability: 1290 Infinity LC- System Emulation Technology

Concept

EmulatedSystem A

FA

Select System to be emulated by a simple mouse click:

Agilent 1100 Series

Agilent 1200 Series LC

Agilent 1260Infinity LC

Agilent 1220Infinity LC Other

Intelligent System emulation technology- How it works (1290 to 1200)

Programmed gradient

1290 gradient

1200 gradient

0 0.5 1 1.5 2 2.5 3 3.50

50

100

150

200

250

300

350

400

min

mAU

Injection

Same gradient conditions by moving the gradient and by emulating the gradient mixing behavior.

July 22, 201121

Intelligent System Emulation Technology ISET- How to?

Select pump

Select autosampler

Chromatographic Evaluation: Conditions

Instruments:(A) 1100 quaternary pump (G1311A) + G1367A autosampler,(B) 1290 Infinity LC system (C) 1290 Infinity system with ISET

Sample: 1 Atrazindesethyl, 2 Hexanzinon, 3 Metoxuron, 4 Cyanazin, 5 Methabenzthiazuron, 6 Chlortoluron, 7 Atrazin, 8 Diuron, 9 Metobromuron, 10 Metazachlor, 11 Sebuthylazin, 12 Nifedipin, 13 Terbuthylazin, 14 Linuron, 15 Prometryn, 16 Nimodipin

Conditions:Gradient: Water-Acetonitrile , 20 – 60% in 8 min,Flow rate: 0.8 ml/min, Column: 2.1x100 mm Zorbax Eclipse Plus, 1.8 µm column.

Method transfer, pesticide example2.1x100 mm Zorbax Eclipse Plus, 1.8 µm column, flow: 0,8 mL/min

July 22, 2011

(C)1290 Infinity

1200 barwith ISET

(A)1100

Quaternary400 bar

(B)1290 Infinity

1200 bar

1 2,3 4 5 6,7 8 9 10,11,12 13 14 15 16

Method transfer, pesticide example Retention times and ΔRT

April 201024

A B1290 Infinity LC

1100 Quaternary

1290 Infinity LC with ISET

PesticidesColumn: 3 x 50 mm, 2.7 µm Poroshell Run time: 3.5 min

25

min1 1.5 2 2.5 3 3.5

mAU

0

100

200

300

400

500

600

700

800

1290 System

Rs= 1.90

1100 Binary System

Rs= 1.15

Sim

azin

e

Chl

orid

azon

e

Chl

orto

luro

n

Diu

ron

Pro

pazi

neTe

rbut

ylaz

ine

Pro

met

ryn

1290 System with iSETM

etam

itron

Cya

nizi

ne

Rs= 1.87


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Intelligent System Emulation Technology ISET- Benefits

Achieve higher productivity for method developmentSpeed up your method development with UHPLC performance and then fine-tune your method by emulating the target system – and be confident that the method will run as intended

Simplify your instrument to instrument method transferNo more method transfer problems! Simply emulate the LC system on which the original method was developed –with a single mouse click.

Minimize your instrument-related costsRun your legacy methods with ISET and at the same time take full advantage of the UHPLC speed, resolution and sensitivity of the1290 Infinity LC. No need to maintain your old legacy LC systems.

Instrument to Instrument Method Transferability- Comparison to current solution?

• From a lower to higher dwell volume LC systemA) Adding physical volume (plumbing solution)B) Modifying your gradient table with isocratic hold

• From a higher to lower dwell volume LC systemC) Delay the injection

Method Transfer Possibilities Today- A) Adding physical volume (plumbing solution)

1290 + 1000 μL dwell vol.

1100/1200 Quat Pump

Example: 1290 Infinity LC

Method Transfer Possibilities Today- A) Adding physical volume (plumbing solution)

1290 + 1000 μL dwell vol.

Example: 1290 Infinity LC

Advantages• Works well for not so fast analysis• No Method change

Disadvantages• Requires hardware change, no flexibility• Hardware change difficult in validated environment• Requires manual determination of the dwell

volume (Pumps with damper the dwell volume is not constant and depends on pressure)

1100/1200 Quat Pump

Method Transfer Possibilities Today- B) Isocratic Hold

Example: 1290 to 1200

Injection

0 0.5 1 1.5 2 2.5 3 3.50

50

100

150

200

250

300

350

min

Isocratic hold: move the gradient

Programmed gradient

1290 gradient

1200 gradient

mAU




Programmed gradient

1290 gradient

1200 gradient

Injection

0 0.5 1 1.5 2 2.5 3 3.50

50

100

150

200

250

300

350

min

mAU


Does not consider the gradient mixing behavior (different curves).



1200 Series LCBinary Pump

1290 Infinity LC+ 860 µl “Delay Volume”





1260 Infinity LC

1290 Infinity LC+ 900 µl hold

Advantages• Flexibile, Isocratic Hold is part of the method

Disadvantages• Works not in all cases• Requires manual determination of the dwell volume/

isocratic hold (Pumps with damper the dwell volume is not constant and depends on pressure)

• Requires modification of the methods (should be avoided in validated environment,but doesn’t require revalidation USP Chapter <621>)

LC Method Transfer Possibilities Tomorrow- Intelligent System Emulation Technology (ISET)

Advantages• Simple and flexible

(select the emulated LC system simply by a mouse click)• No determination of the dwell volume etc. required

(perfectly covers pumps with damper and not constant dwell volume)• Works perfectly in all cases• Requires no modification of the methods

(no conflicts in regulated environment)

Agilent 1290 Infinity LC with ISET inside - The Art of Emulation

Emulates other (U)HPLC instruments – by a simple mouse click

Runs existing (U)HPLC methods– without modifying method or system

Delivers same retention times and peak resolution – for infinitely better method transfer

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Thanks for your attention!!!

Questions?

Presentation Title Arial 28pt Bold Agilent Blue · 1 2009 - Stimuli article „Transfer of...

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