quality by design (qbd) solutions for analytical method ......8 qbd approach for method development...

Quality by Design (QbD) Solutions for Analytical Method Development

A Systematic Approach to

Reducing Variability

Content

• Introduction- Traditional Development and Transfer of Methods

- QbD Approach for Method Development

• Agilent Solutions for QbD Method Development-Agilent Method Development Systems

-Intelligent System Emulation Technology (ISET)

• QbD Method Development Workflow- Screening

- Optimization

- Robustness study, Design of Experiments

- Transfer & Verification

3

Key PointsQuality by Design (QbD) Solutions for Analytical Method Development

• Agilent 1290 Infinity Series Method Development Solutions support

automated method development workflows

• Method transfers from Agilent 1290 Infinity Series UHPC systems to other

(U)HPLC systems are seamless by ISET (Intelligent System Emulation

Technology)

• 1290 Infinity Series Method Development Solutions in combination with

ISET and additional QbD software are supporting fast method development

processes and method transfers to other (U)HPLC systems under QbD

aspects

• Agilent Remote Advisor and ACE are excellent solutions to support method

development workflows under QbD aspects for regulated environments

4

Introduction

• QbD (Quality by Design) is defined in the ICH guideline Q8(R2) http://www.fda.gov/downloads/Drugs/Guidances/ucm073507.pdf

• The ICH guidelines suggest to apply Quality by Design

principles in each step to eliminate risk or failures in drug

development processes

• Analytical method development for a drug is also a process

therefore quality principles from the ICH guideline should be

implemented

(ICH = International Conference of Harmonization of Technical Requirements

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

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

Traditional Development and Transfer of MethodsA chromatographic challenge !

5

± 10%

± 10oC

± 50%

± 10%

± 0.2

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

Fixed method specificationsdetermined manually

Transfer

Transfer

Develop ValidateTransfer

R & DQA / QC

6

Variability in resolution of criticalpeaks from day to day

Different results on different systems

Buffer solutions prepared bydifferent operators deliverdifferent results

Even small changes in pH show a large effect

Traditional Development and Transfer of MethodsReported Issues After Method Transfer

FDA Warning Letter Search for “Stability”Frequently stability issues reported

71 % of HPLC warning letters

Include Reference to “stability”

0 10 20 30 40 50 60 70 80

Calibration

Training

OOS

SSC

Method Validation

Defecient Methods

Data Integrity - Security

Data Integrity - Data Deletion

Data Integrity - Incomplete Records

Reaccurance

External Consultant Required

Data Review

Complete Data

Poor Response / CAPA

Stability

41

27

51

27

43

17

27

13

43

30

16

27

43

43

71

8

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 and to create a list of critical method parameters

(CMPs- flow rate, temperature etc.) and critical method attributes

(CMAs- resolution, peak tailing)

• 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

± 10%

± 10oC

± 50%

± 10%

± 0.2

Slope 14.5% MeOH20mM bufferpH 745oC1.0 mL/min

Method specifications:

Traditional Approach• Fixed Protocol

Design space

QbD Approach• Variable Protocol

Traditional Approach vs. QbD Approach

9

Working within the design space will ensure the method‘s robustness

Content

• Introduction- Traditional Development and Transfer of Methods

- QbD Approach for Method Development

• Agilent Solutions for QbD Method Development-Agilent Method Development Systems

-Intelligent System Emulation Technology (ISET)

• QbD Method Development Workflow- 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

Agilent Method Development Systems

12

• 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

13

1290 Infnity I System 1290 Infnity II System

Autosampler

pump

detector

Multi Column Thermostat

MCT

1290 Infinity II Method Development Solution

Why different separation conditions?

P

a

g

e

1

4

min0.5 1 1.5 2 2.5 3 3.5

mAU

0

10

20

Eclipse plus C18

min0.5 1 1.5 2 2.5 3 3.5

mAU

0

10

20

Eclipse plus PhenylHexyl

min0.5 1 1.5 2 2.5 3 3.5

mAU

0

10

20

Bonus RP

Changes of a have a much higher effect than changes of N !


Aspects of Automated Method Development

15

• maximum method performance

• best combination of selectivity, resolution and speed

Analytical Efficiency

• maximum instrument utilization

• simple setup for method development

• no need for manual system changes

Instrument Efficiency

• maximum automation of routine tasks

• unattended data acquisition

• easy data evaluation

Laboratory Efficiency

link to video

http://www.chem.agilent.com/en-US/Promotions/pages/infinity-method-development.aspx

1290 Infinity II Method Development Solution16

8

x

• 2x external solvent selection valves for up to 24 solvents plus built-in solvent valve for

another 2 solvents = 26 solvents in total

• 1x unique Agilent 8pos/18 port valve for up to 8 columns in a single MCT

• 8x individual heat exchanger in two temperature zones

Up to 1300 bar!

12 + 1

x

12 + 1

= 169

= 1352

1290 Infinity II

MCT

1290 Infinity II

Flexible Pump

External Solvent

Selection Valves

Up to 12 solvents

per SSV!!!

A2A1 B2B1

SSV2SSV1

Have access to > 1000 sets of unique separation conditions!


Product and Concept

17

http://www.chem.agilent.com/Library/brochures/5990-6226EN.pdf

Autosampler

Pump

Detector

New 1290 Infinity II LC

All in a single stack!

Up to 1300 bar!

http://www.chem.agilent.com/Library/brochures/5990-6226EN.pdf

1290 Infinity II Method Development Solutionproduct and concept

J

a

n

u

a

r

y

2

0

1

5

18

18

Infinity II

configuration

configuration

Infinity II modules

Valves… ….located in

comment

1 x 8pos/18port valve(G4239C)

1 x 1290 Inf II MCT (G7116B)

for access to up to 8

columns (column selection)

1 x 12pos/13port valve(G4235A)

1 x 1290 ext valve drive(G1170A)

1 x 12pos/13port valve(G4235A)

1 x 1290 ext valve drive(G1170A)

for access to up to 26

solvents (24 + 2 from pump)

Autosampler

Pump

Detector

Up to 1300 bar!

add column to column data base

1290 Infinity II Method Development Solutionproduct and concept – column centric view

J

a

n

u

a

r

y

2

0

1

5

19

19

one click column selection

20


product and concept – column centric view

• selected column is shown directly in the instrument control

• no need to deal with individual valve positions

January 2015

22

Agilent ChemStation Method Scouting Wizard

new features

p.81 ff, http://www.chem.agilent.com/Library/usermanuals/Public/G4230-90004_MDS_USR_EN.pdf

Feature Description Description

Version MSW A.02.03 / A.02.04 MSW A.02.05

Supported screening pumps • 1290 Infinity Binary Pump (G4220A)

• 1290 Infinity Binary Pump VL (G4220B)

• 1290 Infinity Quaternary Pump(G4204A)

• 1260 Infinity Quaternary Pump (G1311B)

• 1260 Infinity Binary Pump (G1312B)

• 1260 Infinity Bio-inert Quaternary Pump (G5611A)

• 1260 Infinity SFC Binary Pump (G4302A)

• 1290 Infinity Binary Pump (G4220A)

• 1290 Infinity Binary Pump VL (G4220B)

• 1290 Infinity Quaternary Pump(G4204A)

• 1260 Infinity Quaternary Pump (G1311B)

• 1260 Infinity Binary Pump (G1312B)

• 1260 Infinity Bio-inert Quaternary Pump (G5611A)

• 1260 Infinity SFC Binary Pump (G4302A)

• 1290 Infinity II High Speed Pump (G7120A)

• 1290 Infinity II Flexible Pump (G7104)

Support of pump clusters • Pump Valve Cluster with all pumps above

• 1260 Infinity Preparative Pump Cluster

(2x G1361A)

• Pump Valve Cluster with all pumps above

• 1260 Infinity Preparative Pump Cluster

(2x G1361A)

Support of multiple screening

pumps, as e.g. in SFC hybrid

system

If more than one pump is configured, as e.g. with the G4302A

plus LC pump, the user can select which pump will be used in

the screening campaign

If more than one pump is configured, as e.g. with the G4302A

plus LC pump, the user can select which pump will be used in

the screening campaign

Pump specific configuration

parameter

The default parameter used for flushing, equilibration, and

column storage can be stored separately for each pump

The default parameter used for flushing, equilibration, and

column storage can be stored separately for each pump

Supported column

screening configuration• Column compartment cluster of 2 or 3

1290 Infinity Thermostatted Column compartment

(G1316C)

• Column compartment cluster of 2 or 3

1290 Infinity Thermostatted Column compartment

(G1316C)

• Single 1290 Infinity II Multicolumn Thermostat (G7116B)*

with 6/14 or 8/18 valve

Report templates – Intelligent

Reporting

Several report templates are provided to support identification of

most interesting chromatograms

Several report templates are provided to support identification of

most interesting chromatograms

Automatic scheduling of

scouting sequences

The system will automatically schedule a

screening sequence (OpenLAB ChemStation)

The system will automatically schedule a

screening sequence (OpenLAB ChemStation)

*cluster of 2 or more MCTs is planned for later this year

January 2015

http://www.chem.agilent.com/Library/usermanuals/Public/G4230-90004_MDS_USR_EN.pdf

Setup Columns in TCC

(Edit Columns – Online Session Only)

Page 23

Column Setup in TCC

Page 24

Colors Corresponds to

Column Clips and

Tubing Collars

Setup Multi-Channel Valve Solvents

Page 25

Solvent Choices Appear in Pump Setup

Page 26

Method Development Scouting Wizard

Page 27

Choose Root Folder for Campaign

Description May Be Edited

Page 28

Campaign Uses Base Method From Chemstation

Define Screening Campaign Base:

For a method scouting campaign, a base method needs to be set up (you do this as

usual in the ChemStation

It is recommended to use Edit Entire Method

All parameters that are not changed are taken from this base method

These are typically the detector settings, autosampler settings, pump settings that

are not altered, and data processing parameters

Page 29

Choose Base Method & Screening Parameters

Page 30

Choose Columns to Be Used

Note: Column Properties Obtained From Column Table

Page 31

Choose Solvents to be Used

Note: Solvents Choices Obtained from Valve Table

Page 32

Setup Gradient(s) Profile(s)

Page 33

Gradients May be Setup with Timetable or

Graphically

Page 34

Analysis Steps: Optimized for Minimal

Number of Steps

Page 35

Column Temperature Screening

Page 41

Review and Select Methods

Note Methods are Flagged if Outside

Limits (Temp & pH)

Page 42

Exclusion may be overridden

43

Method Scouting Wizard Reporting

All available chromatographic information can be used for

reporting - across-samples, across-sequences, across-labs*!

Tables, diagramms, conditional formating.... are available.

*) across labs - only in combination with OpenLAB ECM

44


Scouting

conditions

sorted for

highest

number of

peaks

Many options for sorting and

filtering....

45


Experiments vs.

Maximum retention

time and maximum

number of peaks

(bubble-size)

Experiments vs.

retention time of

peaks (projection of

all chromatograms)

Many options for graphical overviews,

e.g. ...

46


Data-file, column,

solvents, temperature,

gradient-name from

Method Scouting Wizard

And of course chromatograms – with scouting

conditions!

Intelligent System Emulation Technology (ISET)

47

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

PO-Nr. G2197AA

Method Transfer Between Different LC Instruments

48

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

49

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


50

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


Still gradient differences due to

different mixing behavior

1260 Infinity LC

1290 Infinity LC

+ 900 µl hold

51

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


52

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

Almost consistency of both

gradient curves


1290 + 1000 μL dwell vol.

1100/1200 Quat Pump

Approach # 2: Adding a Physical Void Volume

Results

53

• 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)

54

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

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

55

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

1290 Infinity Quaternary Pump - BlendAssist

Simple tool for online-dilution of modifiers and gradient set-up

You need different concentrations of modifiers in your analysis, would like to have just one stock-

solution and do online dilution to profit from the quaternary mixing capability of your pump? Here is a

simple tool – BlendAssist!

Water Water

1% TFA

ACN

ACN

1% TFA

Desired method conditions - example:

1. 5 to 95% gradient of ACN with 0.1% TFA in

Water and 0.08% TFA in ACN

2. 20 – 80% gradient of ACN with 0.5% TFA in

Water and 0.4% TFA in ACN

Without BlendAssist you need to either pre-mix the

required solvents or by using stock-solutions of

TFA in Water and ACN to program complex

gradients (%A, B, C, D).

With BlendAssist: just program your binary

organic/aqueous gradient and define the dilution

factor!

56

1290 Infinity Quaternary Pump - BlendAssist

Simple tool for online-dilution of modifiers and gradient set-up

57

min1 2 3 4

Norm.

0

5

10

15

20

25

30

35

40

6 dynamically mixed mobile phases

6 premixed mobile phases (same user)

min1 2 3 4

Norm.

0

5

10

15

20

25

30

Analysis of Glucocorticoids - Comparing dynamically mixed mobile phase vs. user influence

58

A:B=70:30

A:B=70:30

Buffer Advisor SoftwareQuaternary mixing

February 3, 201759

Buffer Advisor SoftwareWorkflow

67

High Dynamic Range (HDR)Increasing the linear range by a factor of >30

1260/1290 Infinity DAD

> 30 x Linearity

with HDR

1.4 1.8 2.01.3

>10

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

1040 > 1090 1090 > 1050 1050 > 1100 1100 > 1200 1200 >1290

History of DAD Sensitivity Gain - The last 30 years

Detector Model (Intro Year)

Sensitivity

Gain

(1988)(1986) (1995) (2006) (2010)

1260 / 1290 Infinity

Diode Array Detector

Page 68

History of DAD Linearity Gain - The last 30 years

Detector Model (Intro Year)

(1988)(1986) (1995) (2006) (2010)

Page 69

1.4 1.8 2.0 1.42.9

39.1

0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

1040 to 1090 1090 to 1050 1050 to 1100 1110 to 1200 1200 to 1290/60 1200 to HDR

Linearity

Gain

(2011)

1260 / 1290 Infinity

Diode Array DetectorWith HDR

>30X Linearity

Deuterium

Lamp

Max-Light Cartridge Cell

10 mm or 60 mm path length

Programmable

or fixed slit

1024 element

diode-array

GratingMirror

Optofluidic Waveguides: Max-Light Flow Cells - Total-internal reflection in a non-coated fused silica fiber

1260 / 1290 Infinity DAD

Page 70

Effects of path length increase

Peak Dispersion

60 mm pathlength

large geom. cell volume (78 µL)

lower light transmission

- Loss of resolution

- Loss of signal height

10 mm pathlength

small geom. cell volume (13 µL)

high light transmission

Conventional flow cells:

Page 71

60 mm pathlength

4 µL σV dispersion volume

Max-Light High Sensitivity cell:

Optofluidic waveguides

(total internal reflection)

10x Higher Sensitivity 1290/1260 Infinity DAD compare to 1200 Series DAD SL

1260/1290 DAD

60 mm

1200 DAD SL

10mm

Height [mAU] 28.876 4.938

Noise [mAU] 0.0098 0.0190

Signal/ noise 2944 259

Increase 11.4 x

min0 1 2 3 4 5

mAU

0

5

10

15

20

25

30

Columns: 150 x 4.6mm Zorbax SB C18, 5µm

Sample: Anthracene: 835 pg/µL

Mobile phase: A: Water, B: Acetonitrile

Elution: isocratic 80 % B

Injection volume: 5 µL

Flow: 1.5 mL/min

DAD: 251/4nm, Ref= 450/80nm,

2.5Hz, slit width 4nm

1200 Series

DAD / DAD SL

1260/1290

Infinity DAD

Page 72

Max-Light cartidge cells for 1260/1290 DAD- Specifications

73

10 mm path length 60 mm path length

Max-Light

cartidge cell

Path

length

Dispersion

volume σv

Noise Noise

per cm

Standard 10 mm 1.0 μL ± 3.0 µAU ± 3.0 µAU/cm

High Sensitivity 60 mm 4.0 μL ± 3.6 µAU ± 0.6 µAU/cm

Linearity, linear range and dynamic range- 1200 Series Diode Array Detector, 10 mm flow cell

Page 74

Conc [µg/mL]

Detector Signal

[AU]

LOD

(3x S/N)Conc (upper limit)

linear range

Detector

noise

7*10-6

Conc (upper limit)

dynamic range

Linear range

Dynamic range

2.0

Slope = Response Factor

(Sensitivity)

1260/1290

DAD

1260/1290

DAD

Upper limit

5

10_3

__

NoiseDetector

LimitUpperrangeLinear

10 mm

Linearity, linear range and dynamic range- 1260/1290 Infinity Diode Array Detector, 10 mm flow cell

Page 75

Conc [µg/mL]

Detector Signal

[AU]

LOD


linear range

Detector

noise

3*10-6

Conc (upper limit)

dynamic range

Linear range

Dynamic range

2.5

typically

~ 3 times higher linear range vs. 1200 Series DAD

(due to lower detector noise, higher upper limit)

10 mm

Linearity, linear range and dynamic range- 1260/1290 Infinity Diode Array Detector, 60 mm flow cell

Page 76

Conc [µg/mL]

Detector Signal

[AU]

LOD

(3x S/N)

6 times higher signal

Conc (upper limit)

linear range

Detector

noise

3.6*10-6

Conc (upper limit)

dynamic range

Linear rangeDynamic range

2.5

typically

Shift of linear and dynamic range to lower concentration,

but same linear range: Gain at lower concentration,

loss at higher concentration)

Combine two flow cell

with short and long path length

60 mm

Linearity, linear range and dynamic range- 1260/1290 Infinity DAD with HDR (3.7 mm and 60 mm flow cell)

Page 77

Conc [µg/mL]

Detector Signal

[AU/cm]

LOD


linear range

Detector

noise

0.6*10-6

Conc (upper limit)

dynamic range

Linear range

Dynamic range

6 .7

60 mm

Flow cell 3.7 mm

Flow cell

+One normalized signal AU/cm resulting in linear range

> 3.7 x 106 (39x higher than 1200 Series DAD)60 mm 3.7 mm

78

3.7mm

60mm

Clustered DAD

1290/1260 High Dynamic Range – HDR Increasing Linear UV Range by >30x

Technology:

Cluster of two DADs

• DAD 1:

3.7mm Cell – for high concentrations

• DAD 2:

60 mm Cell – for low concentrations

• Output: One, combined signal,

normalized to 10 mm path length

(HDR range: 0.6 x 10-6 to 6.7 AU/cm)

vs for 1200 Series: 7 x 10-6 to 2 AU/cm)

Control / Usability

• Like standard 1290/1260 DAD:

Control, data analysis, reporting

Investment Protection

• Existing 1290/1260 DAD can be upgraded

(by 2nd DAD)

• Existing 1100/1200 Systems can be

upgraded (by two 1290 or 1260 DAD)

DAD 1:

DAD 2:

High Dynamic Range (HDR) Signal

Page 79

The combined signal is

dominated by the long path

The combined signal is dominated by the short path

Transition range

Impurity Analysis, DAD with 10 mm flow cell

February 3, 2017Confidentiality

Label

81

min2 3 4 5 6 7

mAU

0

2

4

6

8

0.07 %

S/N 21

0.63 % 0.02 %

S/N 3.2

0.68 %

0.01 %

S/N 1.4

3.83 %

0.002 %

S/N 0.4

9.99 %

0.05 %

S/N 14

0.70 %

min2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8

mAU

0

500

1000

1500

2000 Impurity level

Signal/Noise

Area rsd %

Impurity Analysis, DAD with 60 mm flow cell


Label

82

min2 3 4 5 6 7

mAU

0

2

4

6

8 0.07 %

S/N 52

0.38 % 0.02 %

S/N 7.9

0.29 %

0.01 %

S/N 3.6

0.90 %

0.002 %

S/N 0.9

7.92 %

0.05 %

S/N 35

0.35 %

min2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8

mAU

0

500

1000

1500

2000

2500

3000

3500

Impurity level

Signal/Noise

Area rsd %

Improved S/N and RSD

Main not within linear range

Impurity Analysis, DAD HDR


Label

83

min2 3 4 5 6 7

mAU

0

2

4

6

8

0.07 %

S/N 53

0.35 % 0.02 %

S/N 8.0

0.30 %

0.01 %

S/N 3.6

1.57 %

0.002 %

S/N 0.9

11.1 %

0.05 %

S/N 35

0.35 %

min2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8

mAU

0

250

500

750

1000

1250

1500

1750

2000

Impurity level

Signal/Noise

Area rsd %

Improved S/N and RSD

Main within linear range

Impurity Analysis, DAD HDR, 3x Inj. Vol. (3µL)


Label

84

min2 3 4 5 6 7

mAU

0

2

4

6

8

0.07 %

S/N 159

0.30 % 0.02 %

S/N 29

0.28 %

0.01 %

S/N 13

0.48 %

0.002 %

S/N 3.2

4.88 %

0.05 %

S/N 108

0.28 %

min2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8

mAU

0

500

1000

1500

2000

2500

3000

3500

Impurity level

Signal/Noise

Area rsd %

Further Improved S/N and RSD

Main still within linear range

87

1290/1260 High Dynamic Range – HDR Increasing linear UV Range by 30x

AdvantagesImpurity Analysis

• Main + impurities in one run

• 30x lower LOD/LOQ (vs 1200)

• 15x lower LOD/LOQ (vs 1290/1260)

• 3 x higher upper limit of linearity

• ID / Quant of lower level

impurities (0.002% / 0.01%)

• Ideal for fixed dose combinations

• Prepared for stronger regulations

• Genotoxins

Improved Area Precision enabling

Automated Integration

• Higher Productivity

• Less error-prone

• Higher data quality and security

3.7 mm

60mm

Clustered DAD

ChromSword

AutoChrom (ACD Labs)

Fusion QbD (S-Matrix)

Add-on Software Options for QbD Method Development

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

90

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.).

February 3, 201791

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

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

93

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

94

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

95

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

96



namely

• Step # 1: Screening

• Step # 2: Optimization

Screening


• pH conditions



Agilent Method

Scouting

WizardOptimization

• Optimize Gradient Profiles

• pH conditions

• Flow rates, temperaturesQbD Software


97

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

99



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


3D Graph Called

Response

Surface

Example - Why QbD is Needed…..Step # 3: Design of ExperimentsInvestigate multivariate relationships

Center Point of

Robust region –

Prediction

Center Point of

Robust region –

Experiment

API-Symmetry 0.71±0.04 0.66

API-tailing USP 1.4±0.07 1.4

API-Tangent

Width

0.081±0.004 0.083

API-Resolution 2.3±0.3 2.0

Optimized Parameters

Moving critical pair in

a more robust region

101

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.

102



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


103

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

104

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

105

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

106

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

Agilent Application Note

February 3, 2017107

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

108

*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

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, Blend Assist, Buffer Advisor, and

third party QbD software provides a unique and efficient way

to develop and transfer robust methods

109

February 3, 2017

Confidentiality Label

110

THANK YOU

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

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

112

quality by design (qbd) solutions for analytical method ......8 qbd approach for method development...

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