End Market Training Template FY13

1

Quality by Design (QbD) Solutions for Analytical Method Development

Andreas Tei

Pharmaceutical Segment Manager

Life Sciences Group

A systematic approach to reducing

variability

Content

• Introduction- Traditional approach of method development and transfer

- QbD approach for method development

• Agilent solutions for method development-Agilent method development systems

-Intelligent system emulation technology (ISET)

-Method scouting wizard software

-Third party QbD software

• A practical approach of method development under QbD principles- Screening

- Optimization

- Robustness study, Design of Experiments

- Transfer & Verification

2

3

Introduction

• Analytical method development is an important part of the drug

development process therefore the quality principles which

have been described in the ICH guideline Q8 (R2) should be

implemented to eliminate risks or failures

• http://www.fda.gov/downloads/Drugs/Guidances/ucm073507.pdf

(ICH = International Conference of Harmonization of Technical Requirements

for Registration of Pharmaceuticals for Human Use, Founded in 1990 by an FDA initiative)

Traditional Development and Transfer of MethodsA chromatographic challenge !

4

± 10%

± 10oC

± 50%

± 10%

± 0.2

Slope 14.5% min MeOHpH 735oC1.0 mL/min

Fixed protocol describingChromatographic conditions

Transfer

Transfer

Develop ValidateTransfer

R & DQA / QC

http://www.fda.gov/downloads/Drugs/Guidances/ucm073507.pdf

3

5

Variability of critical methodattributes (resolution, tailing, etc)

Small changes in pH, temperatureor flow rate shows a large effect

Method Transfer is a balancing act

10% of analytical methods are discarded per year to avoid high revalidation costs

after the methdod showed instabilities on the QC system

Development

System

Target QA/QC

Systems

Even buffer solutions from different

operators deliver different results

Different results on different systems

6

QbD Approach for Method Development

• Analytical QbD begins by defining goals (Analytical Target

Profile, ATP) and identifying potential method variables and

responses that affect method quality

• Statistical „Design of Experiments“ (DOE) has been applied to

the selected method variables leading to process and method

understanding

4

7

QbD Approach for Method Development

• A list of critical method parameters (CMPs- flow rate,

temperature etc.) and critical method attributes (CMAs-

resolution, peak tailing) will be worked out

• The experimentally measured responses were modeled to

determine the design space (defined in ICH Q8 (R2))

• A verified and validated method can be modified within the

design space to compensate any unforseen variables yet

delivering consistent results

Quality by Design - A systematic approach to obtain a“consistent quality” output

System Variabilities

Fixed process

Variable output

System Variabilities

Flexible

process

Consistent output

Quality by

Design

Understand variability

Continuous improvement

Control Variability

ICH Q8-Q11

ICH = The International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use

May 18, 2015

8

5

Content

• Introduction- Traditional approach of method development and transfer

- QbD approach for method development

• Agilent solutions for method development-Agilent method development systems

-Intelligent system emulation technology (ISET)

-Method scouting wizard software

-Third party QbD software

• A practical approach of method development under QbD principles- Screening

- Optimization

- Robustness study, Design of Experiments

- Transfer & Verification

Agilent Solutions for QbD Method Development

ISET

Intelligent System Emulation Technology

RA

Remote Advisor

Harmonized Qualification

ACE

Method Development

System & Method Scouting SW

6

Agilent Method Development Systems

11

• 1352 different combinations of column chemistries and eluents

• A nearly infinite number of separation conditions is created by including

different temperature and flow rates as variable parameters

1290 Infinity II Method Development SolutionNew: Multi Column Thermostat

12

1290 Infnity I System 1290 Infnity II System

Autosampler

pump

detector

Multi Column Thermostat

MCT

7

1290 Infinity II Series Method Development Solution

New: Column Centric View

13

one click column selection

Unequivocal column selection in the instrument method for higher confidence

Intelligent System Emulation Technology (ISET)

14

- Seamless transfer of methods between LCs, regardless of the brand

PO-Nr. G2197AA

8

Method Transfer Between Different LC Instruments

15

min3 4 5 6 7 8 9 10 11

mAU

0

25

50

75

100

125

150

175

1100 Series Binary System

1290 Infinity LC

Method transfer from a UHPLC system with a minimized dwell

volume and optimized mixing behavior to any other LC system is

often challenging and affects rentention time and resolution

Example: Method Transfer from a 1290 UHPLC to a 1100 HPLC system

May 18, 201516

Practical aspects how to measure dwell volumes, transfer & optimize methods

9

17

1290 Infinity LC

min0 1 2 3 4 5 6 7 8 9

mAU

0

100

200

300

400

500

600

1100 Series Quaternary LC

Gradient differences due to different dwell

volumes and different mixing behavior

Method Transfer Between Different LC Instruments

Results from a tracer experiment

min0 1 2 3 4 5 6 7 8 9

mAU

0

100

200

300

400

1290 Infinity LC

1 min isocratic hold


18

Method Transfer Between Different LC InstrumentsApproach # 1: Isocratic holding step to synchronize


Still gradient differences due to

different mixing behavior

10

1260 Infinity LC

1290 Infinity LC

+ 900 µl hold

19

Approach # 1: Applying Isocratic Holding Steps

Results

• Results shows a low consitency

• Requires manual determination of the dwell volume/ isocratic hold

(in solvent delivery systems equipped with dampeners the dwell volume is pressure

dependent and variable)

• Requires modification of the methods

(should be avoided in validated environment,

but doesn’t require revalidation USP Chapter <621>)

min0 1 2 3 4 5 6 7 8 9

mAU

0

100

200

300

400

1290 Infinity LC

1 mL loop installed


20

Method Transfer Between Different LC InstrumentsApproach # 2: Adding a physical void volume

Almost consistency of both

gradient curves


11

1290 + 1000 μL dwell vol.

1100/1200 Quat Pump

Approach # 2: Adding a Physical Void Volume

Results

21

• Results shows a good consitency

• Manual determination of dwell volumes required (issues of a variable

dwell volume with systems containing damperners)

• All mechanical changes are laboriously not flexible

Agilent Solution:

Intelligent System Emulation Technology (ISET)

22

Programmed gradient

1290 gradient

1200 gradient

0 0.5 1 1.5 2 2.5 3 3.5

0

50

100

150

200

250

300

350

400

min

mAU

InjectionSoftware controlled compensation

of dwell volume differences and

synchronization of mixing

behaviors

12

Agilent Solution: Method Transfer by ISETResults: 1260 Infinity Binary LC to 1290 Infinity LC

23

min1 2 3 4 5 6 7 8 9

mAU

0

10

20

30

40

Imp

A

Imp

B

Imp

H Imp

J

Imp

F

Imp

K

Pa

race

tam

ol

1290 Infinity LC with ISET

1260 Infinity Binary LC

• Consistency of results

24

Automated Method Development SoftwareAgilent Chemstation Method Scouting Wizard Software

• Define project

Choose scouting combinations and

base method.

• Select columns

All installed columns are shown automatically.

• Select solvents

Pump types and valves are automatically

detected.

• Define gradients

Select between different gradients and

temperatures.

• Review and select screening methods

Check for incompatible

combinations.PO-Nr.

G2196AA

13

Method Transfer from UHPLC to HPLC

May 18, 201525

HPLC Calculator v3.0

http://www.unige.ch/sciences/

pharm/fanal/lcap/telechargement.htm

Agilent Method Translator

www.agilent.com/chem/1200calculator

Crawford Scientific Calculatorhttp://www.crawfordscientific.com/

HPLC_Method_Transfer_On-line.htm

ChromSword

AutoChrom (ACD Labs)

Fusion QbD (S-Matrix)

Add-on Software Options for QbD Method Development

http://www.unige.ch/sciences/

http://www.agilent.com/chem/1200calculator

http://www.crawfordscientific.com/HPLC_Method_Transfer_On-line.htm

http://www.crawfordscientific.com/HPLC_Method_Transfer_On-line.htm

14

4.5

Rs = 9.3 + 4.2(∆T) - 5.4(%I)2 + 12.7(pH)2 + 1.3(∆T *pH) + 1.6(∆T)2%I+ …

Linear Effects Curvature

Effects

Complex EffectsInteraction

Effects

pH

Oven Temp

Initial %

Organic

5.0

15.0

30.0 60.0

27

2.5

3.5

Fusion QbD Software (S-Matrix)

• Uses a DOE and statistical modeling approach.

• Example equation below illustrates resolution is a complex function of different

interacting parameters (temp, flow, pH, etc.).

May 18, 201528

Details of Fusion QbD Software (S-Matrix)Variable ConstantColumn (Agilent ZORBAX RRHD Eclipse Plus

Phenyl-Hexyl) length

• Gradient

Equilibration : 1.0 min

5%B

Initial hold: 1 min, 5% B

Final Hold: 2 min at

95%B

Re-equilibration: 4min

at 5%B

• Flow Rate: 0.6 mL/min

• Injection volume: 1 µL

• Wavelength: 245 nm ±

4 nm (ref off)

• All other finalized

parameters from

Phase 1

• 3.0x50 mm, 1.8 µm (p/n: 959757-312)

• 3.0x100 mm, 1.8 µm (p/n: 959964-312)

Solvents

• A1: pH 4.0, 5 mM formic acid and 10 mM

ammonium formate in water

• A2: pH 4.5, adjusted from pH 5 with

acetic acid

• A3: pH 5.0, 5 mM acetic acid and 10 mM

ammonium acetate in water

• A4: pH 5.5, adjusted from pH:7 with

acetic acid

• A5: pH 6.0, adjusted from pH:7 with

acetic acid

• A6: pH 6.5, adjusted from pH:7with acetic

acid

• A7: pH 7.0, 10 mM ammonium acetate in

water

• B1: Acetonitrile

• B2: Acetonitrile: THF (88:12)

Gradient time

• 9 min

• 15 min

Column temperature

• 35°C

• 40°C

• 45°C

• 50°C

Response Goals Target Lower

Bound

Upper

Bound

Relative

Rank

No. of Peaks Maximize 36 45 1

No. of Peaks >= 1.50 (Tangent

Resolution)

Maximize 28 34 0.9

No. of Peaks >= 2.00 (Tangent

Resolution)

Maximize 21 28 1

Max Peak #1 (Tangent Resolution) 2.51 1.40 3.62 1

Max Peak #1 (Symmetry) Minimize 1.14 1.15 0.9

left side:

Example of Parameters used to optimize

methods

bottom:

Example: Trend Response goals used in

the chemistry screening phase

15

List of CMPs and CMAs for robustness

testing

* The coded names are used in robustness model displays

Details of Fusion QbD Software (S-Matrix)Robustness Testing & Establishing Design Space (MODR)

Design Space

May 18, 201530

Details of Chromsword Software

• Method development and optimization based on

the solvatic model of RP-LC

• Building retention models after processing results

by applying different variables as listed below

• Organic modifier concentration (C)

• Gradient time (Grad)

• Temperature (T)

• pH

• Multi variables: C+pH; Grad +T; C1 +C2

16

May 18, 201531

min0.5 1 1.5 2 2.5 3 3.5 4 4.5

Norm.

0

100

200

300

400

500

600

1.9

70

2.0

49

2.8

20

3.0

70

min1 2 3 4 5 6 7 8 9

Norm.

0

20

40

60

80

100

1.1

36

1.3

84

3.2

32


• Automated resolution optimization of critical peak pairs

May 18, 201532


Design space creation after robustness study is fundamental to the

Quality by Design approach

17

May 18, 201533

Chromsword Software

• Interactive

• Sample profiling

• Impurities search

• Full factorial

• Statistical DOE

• Interactive

• Find workable method ASAP

• Method scouting

• User defined

• Statistical DOE

ScreeningRapid

optimization

Fine Optimization

Robustness Studies

5/18/201534

4. Modeling

Temperature

Gra

die

nt

ACD/AutoChrom Software

Start

Column, Buffer &

Solvent Screening

Select Best

Optimize Gradient & Temperature

End

1. Data Collection

2. Peak Tracking

5. Next

Experiment

Report

3. Interpretation3. Interpretation

18

Content

•Introduction

– Traditional approach of method development and transfer

– QbD approach for method development

• Agilent solutions for method development–Agilent method development systems

–Intelligent system emulation technology (ISET)

–Method scouting wizard software

–Third party QbD software

• A practical approach of method development under QbDprinciples

– Screening

– Optimization

– Robustness study, Design of Experiments

– Transfer & Verification

36

31

Target System

Emulation by ISET

Robustness study by

QbD software

1290

R&D

1260QA/QC

Method transfer

Method development

System

Use of 1.8 µ particles

and QbD software

Target Systems in

QA/QC labs

QbD Method Development & Method Transfer WorkflowFrom UHPLC to HPLC in a nutshell

2

1290

R&D

1260

R&DEmulation

19

37

Overall workflow which

consists of four main steps

namely

• Step # 1: Screening

Screening

• Column Chemistry

• pH conditions

• Organic Solvent Selection

• Gradients, temperatures

Agilent Method

Scouting

Wizard

QbD Based Method Development Workflow

38

Step # 1: ScreeningResults

For more details please see Agilent Application Note 5991-0989EN

Bubble size represents the number of integrated peaks and, consequently, best

mobile and stationary phase combination

20

39



namely

• Step # 1: Screening

• Step # 2: Optimization

Screening


• pH conditions



Agilent Method

Scouting

WizardOptimization

• Optimize Gradient Profiles

• pH conditions

• Flow rates, temperaturesQbD Software


40

Variable Parameters Study Range

Pump Flow rate (mL/min)

Intermediate hold time (min)

Final % Strong Solvent (Gradient 1)*

Oven Temperature (°C)

0.550, 0.600,0.650

3.00 <= Intermediate Hold Time <= 7.00

30.0 <= Final % Strong Solvent <= 35.0

33.0, 36.0, 39.0

Constant Parameters Constant Value

Column Type

Wavelength

Strong Solvent type

pH

Injection Volume

Equilibration Time

Initial Hold Time

Gradient 1 Time*

Gradient 2 Time*

Final Hold Time

Final Hold % Organic

Initial % Strong Solvent

3.0X100 mm,1.8 µm ZORBAX RRHD Eclipse Plus Phenyl-Hexyl

245 nm ± 4 nm (ref off)

Acetonitrile

7.0

1µl

2.50 min

1.00 min

5.17 min

9.28 min

2.00 min

90.0 %B

5.0% B

Step # 2: OptimizationFusion AE QbD Software

Gradient 1: 5%B-(30-35)%B

Gradient 2: (30-35)%B-90%B

Optimization of flow rate, gradient slope, pH, column temperature

21

min10 10.2 10.4

998 Set by User

| || |' ' ' ' '

+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

min10.5 10.55 10.6 10.65 10.7

998 Set by User

| || |' ' ' ' '

++++++++++++++------+++++++++----------+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

min2.5 5 7.5 10 12.5 15 17.5 20

mAU

0

50

100

150

200

250

300

350

10

.54

3

min2.5 5 7.5 10 12.5 15 17.5 20

mAU

0

50

100

150

200

250

300

350

10

.00

7

Step # 2: OptimizationResults: peak purity and separation after optimization 99.8%

99.8 % purity

Before

Optimization

After

Optimization

41

Purity< 90%

42



namely

• Step #1: Screening

• Step #2: Optimization

• Step #3: Robustness study

Screening


• pH conditions



Agilent Method

Scouting

WizardOptimization


• pH conditions


QbD Software

Design of Experiments

• Multivariate study

• Robust region

• Design Space


22

43

Step #3 : Design of Experiments

Results: Design SpaceCritical Method

Parameters (CMPs)

Proven

Acceptable

Range (PARs)

Critical Method

Attributes (CMAs)

Column: Agilent ZORBAX

RRHD Eclipse Plus C8 3.0X50

mm, 1.8 µm

No. of peaks (>40)

API resolution (>1.5)

Peak purity (≥ 98%)

Peak tailing (<1.5)

Strong solvent: Methanol

% Strong solvent: 90.5% ± 1.5%

Aqueous solvent pH: 7.7 ± 0.1

Gradient range: 5% to 90.5%

Oven Temperature: 45°C

Gradient time: 15 min

Flow rate: 0.6 mL/min

Wavelength: 292 nm

CMAs to create a Design Space

The Design Space is a region in which changes to method parameters will not

significantly affect the results.

44



namely

• Step #1: Screening

• Step #2: Optimization

• Step #3: Robustness study

• Step #4: Method Transfer

& Verification

Screening


• pH conditions



Agilent Method

Scouting

WizardOptimization


• pH conditions


QbD Software

ISET

QbD Software

Design of Experiments

• Multivariate study

• Robust region

• Design Space

Method Transfer &

Verification

• Agilent Method Translator

• ISET

• QbD Optimization

• Verification


23

45

3

Target System

Emulation by ISET

Robustness study by

QbD software

1260QA/QC

Method transfer

Target Systems in

QA/QC lab

Verification

Step # 4: Method Transfer & VerificationFrom UHPLC to HPLC

2

1290

R&D

1260

R&DEmulation

46

HPLC design space with new CMA

Robustness Study After Transfer Modeling a HPLC design space on an emulated 1260 system

Critical Method

Parameters (CMPs)

Proven

Acceptable

Range (PARs)

Critical Method

Attributes (CMAs)

Column: Agilent ZORBAX

Eclipse Plus C8 4.6X150

mm, 3.5 µm

No. of peaks (>40)

API resolution (>4)

Peak purity (≥ 98%)

Peak tailing (<1.3)

Strong solvent: Methanol

% Strong solvent: 87.5.% ± 1.5%

Aqueous solvent pH: 7.7 ± 0.1

Gradient range: 5% to

87.5%

Oven Temperature: 37°C

Gradient time: 45 min

Flow rate: 1.4 mL/min

Wavelength: 292 nm

HPLC design space parameters

24

47

Proof Of Robustness After TransferConditions applied from center point and the four corner points of the Design Space

min10 20 30 40 50

mAU

050

150

250

350

min10 20 30 40 50

mAU

050

150

250

350

min10 20 30 40 50

mAU

050

150

250

350

min10 20 30 40 50

mAU

050

150

250

350

min10 20 30 40 50

mAU

050

150

250

350

% B max: 86 %; pH: 7.6

% B max: 89 %; pH: 7.6

% B max: 87.5 %; pH: 7.7

% B max: 89 %; pH: 7.8

% B max: 86 %; pH: 7.8

A

B

D

T

C

API Rs> 4 and API tailing = 1.2 for all runs

Critical Method Attributes remain within the limits

Verification Of The Final Method With The Target SystemResults: 1260 Infinity data compared to 1290 Infinity data in emulation mode

48

Final gradient

Time %B

0.3 5

45.6 87.5

49.2 87.5

49.5 95

49.8 95

50.1 5

53.1 5

mAU

24.0

91

API Rs = 4.2

API tailing = 1.2

min10 20 30 40 50

0

100

200

300

400

23.3

44

1290 emulated as 1260

mAU

100

200

300

400

min10 20 30 40 50

API Rs = 4.1

API tailing = 1.2

0

23.3

47

24.1

15

1260 target system

Critical Method Attributes are within the defined limits

25

Agilent Application Note

May 18, 201549

Title: QbD Based Method Development on an

Agilent 1290 Infinity UHPLC system Combined

with a Seamless Method Transfer to HPLC

Using Intelligent System Emulation Technology

Type: Application Note

Publication Number: 5991-5701EN

Pages: 8

Target segments: Pharmaceutical QA/QC

Language: English

Author: Vinayak A.K.

LitStation Availability: May 5, 2015, CPOD

50

*An extra HPLC optimization time is added after the method transfer to conventional particle sizes to optimize CMAs.

Experiments Sub-2-µm columns

(time in hours)

Conventional columns

(time in hours)

Screening 20 47

Optimization* 28* 20

Total 48 67

Conclusion

• Due to shorter runtimes when using sub-2-micron columns the efficiency

of the method development process has been more than doubled

• ISET can be used to emulate seamless different target systems

• QbD software will deliver robust methods where the critical method

attributes remain constant

• No need for revalidation of methods when applying CMP which are

within the design space

26

Overall Summary

• QbD is a state of the art approach to remove variability of

analytical methods

• ISET enables cross platform method development workflows

• Method Scouting Wizard Software enables easily to create

sequence tables for method development workflows

• The combination of Agilent 1290 systems, ISET, Method

Scouting Wizard Software and third party QbD software

provides a unique and efficient way to develop and transfer

robust methods

51

May 18, 2015

Confidentiality Label

52

THANK YOU

27

QbD process terminology Analytical QbD

terminology

Examples

Analytical Target Profile

(ATP)

Accurate quantitation of API

without interferences from

degradants

Quality Target Product

Profile (QTPP)

Quality Target Method Profile

(QTMP)

pKa, Log P, Solubility

Critical Process Parameters

(CPP)

Critical Method Parameters

(CMP)

Flow rate,

Temperature, pH

Critical Quality Attributes

(CQA)

Critical Method Attributes

(CMA)

Resolution, Peak

Tailing, Peak Capacity

Control Strategy pH ± 0.1; Wavelength

± 2 nm

QbD terminology in Method DevelopmentAppendix

Analytical QbD Terminology

54

• Fast screening of mobile and stationary phases with the Agilent 1290 Infinity LC

and seamless method transfer to an Agilent 1200 Series LC using ISET

Agilent Application Note 5991-0989EN

• Developing faster methods for generic drugs within USP <621>allowed limits


• Effective use of pharmacopeia guidelines to reduce cost of

chromatographic analysis


• Developing faster methods for generic drugs within EP 2.2.46E allowed limits


Appendix: Agilent Application Notes

Method Transfer by ISET

28

Appendix : Agilent Application Notes

QbD based Method Development

• Quality-by-Design Approach to Stability Indicating Method

Development for Linagliptin Drug Product

– Agilent Application Note 5991-3834EN

• Automated QbD Based Method Development and Validation of

Oxidative Degraded Atorvastatin


• Development of an UHPLC Method for Azithromycin Tablets Using

ChromSword Auto Software


• QbD Based Method Development on an Agilent 1290 Infinity UHPLC

system Combined with a Seamless Method Transfer to HPLC Using

Intelligent System Emulation Technology

- Agilent Application Note 5991-5701EN

55

Documents

End Market Training Template FY13