Automated Method Development With UPLC and Fusion Software ... · Automated Method Development With UPLC and Fusion Software A Quality by Design-based Tool ©2012 Waters Corporation

©2012 Waters Corporation 1

Jean-Michel Plankeele

May 2012

Automated Method Development With UPLC and Fusion Software A Quality by Design-based Tool


Agenda

LC method development and Quality-by-Design guidelines

– Definition of key words: QbD, DOE, Design Space…

– Waters’ consistent approach for efficient method

development

FusionMD for method development

– How it works, what it does

Summary & conclusions


Quality By Design Published Guidelines

QbD approach has been there since decades in all

types of industry

More recently implemented in pharmaceutical area

(early 2000)

Guidelines are published by ICH*

Everything about QbD, design of experiment can be

found in ICH Q8 (R2)

– QbD, DoE, design space, graphical tools

Additional details in ICH Q9 & Q10

*ICH: International Conference on Harmonisation


Quality by Design: A systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management

ICH Q8(R2) Guidelines

Some Definitions What Is “Quality By Design”?

QbD is a systematic approach to development …

– No speculation or a priori statements about results

… that begins with predefined objectives…

– Defining criteria (response factors) to be used to measure the

quality of the method

…sound science and quality risk management…

– Statistical treatment of data to provide quantitative values to

goals and limits of response factors

QbD approach is implemented with an experimental design


ICH Q8 (R2) guidelines for Design Of Experiment (DoE)

A structured and organized method of gathering empirical knowledge

DoE are represented as a multi-dimensional spaces (matrix)

Selectivity parameters (study variables)

are the dimensions of this space

Number of experiments is optimized

using an appropriate study design

(full factorial design & partial

factorial design)

Formal Experimental Design: A structured, organized method for determining the relationship between factors affecting a process and the output of that process. Also known as “Design of Experiments”.


Best method

X (pH)

Z (Organic Solvent)

Y (Gradient Time)


What Is a Design? Full Factorial, Partial

With a full factorial design, all elements (experiments) of the

matrix are sampled

In an incomplete factorial design the multidimensional space

is sampled evenly and efficiently

– Factor levels are chosen randomly and then balanced to

achieve uniform sampling

Three variable full factorial design Three variable partial factorial design


ICH Q8 (R2) guidelines Design Space

Around the best method, there is a space where the method achieves

the predifined objectives

This is the area of robustness

Each method (combination of

parameters) within the design

space meets the requirements

Working within the design space

is not considered as a change

– No need for revalidation within

the design space

Design Space: the multidimensional combination and interaction of input variables (e.g., material attributes) and process parameters that have been demonstrated to provide assurance of quality. Working within the design space is not considered as a change


Operating Space

Z optimum

Y optimum

Z

Y (Gradient Time)

X optimum

X (pH)


ICH Q8 (R2) Recommandations For Depiction Of Interactions And Design Space

Response graphs:

– 3 D plots for viewing effects (linear effects, combined effects,

interactions …)

– 3 D plots for evaluating the performances

Contour plots (or 2 D plots) for evaluating the robustness


How Does It Translate When Developping LC Methods?

Quality by design

– Chromatographers must consider all selectivity parameters

– The impact of parameters (range/effect) must be evaluated

– Chromatographers must work with preset objectives

o Cleaning validation Speed

o Impurity profiling Resolution

o Quality control Speed, specificity

Experiment design, design of experiment (DoE)

– Definition of experimental region, selection of study variables with their ranges

Primary selectivity parameters

– Stationnary phase, pH & solvent

Secondary selectivity parameters

– Temperature, flow rate, slope, gradient time, …


Agenda

LC method development and Quality-by-Design guidelines

– Definition of key words

– QbD

– DOE, Design Space…

FusionMD for method development

– What is it, What it does, how it works

Summary, conclusions


What Is Fusion - What It Does

Fusion works in cooperation with Empower I, II & III

– From workstation to network

Fusion supports all UPLC configurations

– ACQUITY UPLC and H-CLASS

– All optical detectors

– Solvent switching valves (external/UPLC or internal/SSV)

– Column managers (temperature control and column

switching)

Fusion supports all Empower controlled configurations

– Alliance, …

Automatically builds methods, method sets and sample

sets


Fusion Alignment With [QbD] Approach

Screening/scouting of all selectivity parameters

Supports RP, NP, IEX, SEC, HILIC, …

Controls all UPLC configurations

All options (detectors, valves, ovens)

Applies the DoE approach

– Structured, optimized set of experiments

– Full factorial and partial factorial designs

Automatically creates and exports all methods

Builds the models of variable effects

Finds the best method

Incorporates robustness modeling in the MD process

[3] Establish Design Space

Define optimum conditions

Define robust operating space

[2] Develop Knowledge Space

Conduct experimental design

Analyze results and study variable effects

[1] Define Experimental Region

Select study variables

Define study ranges


Fusion Workflow In A Few Simple Steps

1. User selects the experimental parameters (study factors or

study variables): Columns, temp, solvents, pH, slope, …

Quality by Design: A systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management



Step 1 Define The Variables And Their Ranges

Flow rate Gradient shape (time, slope, hold, wash, …) Gradient curve Column temp. pH Buffer strength Organic solvent Column type Injection volume Wavelength …





2. Fusion generates all required methods and creates the

experiment design in Empower

Formal Experimental Design: A structured, organized method for determining the relationship between factors affecting a process and the output of that process. Also known as “Design of Experiments”.



Step 2 Create & Export The Design To Empower

Fusion automatically selects the most statistically appropriate

design type and generates the experiment

Minimizes the number of runs

Automatically reconstructs experimental design within Empower

The newly created sample set can be edited/customised

Creates instrument methods and method sets within Empower







3. The methods defined above are run on the system

• Few hours with UPLC vs days with HPLC

…








4. Fusion imports results from Empower and builds the

models


KNOWLEDGE SPACE –

“The information and knowledge gained

from pharmaceutical development

studies and manufacturing experience to

support the establishment of the design

space, specifications, and manufacturing

controls.” [ ICHQ8(R1)]

• Conduct formal experimental design

• Analyze results, build equations

• Run and Process …

Develop

Knowledge

Space*

2

Step 4 Process Results

Rs = βo + β1(X1) + β11(X1)2 + β2(X2) + β22(X2)

2 + β12(X1*X2)

Linear Effect

(main effects term)

Curvature Effect

(simple curvilinear effects)

Interaction Effect

(two-way effects)




study variables): Columns, Temp, solvents, pH, slope, …




4. Fusion imports results from Empower and builds the

models

5. Fusion combines the mathematical models to predict the

optimum method


Step 5 – Automated Numerical Search for Optimum

User enters chromatographic performance goals:

Any calculated result can be selected

Result fields: No. of peaks, No. of peaks > n%, …

Peak fields: Rs of max peak, RT of last peak, purity index of max peak, …

Custom fields can also be selected

Fusion MD identifies study parameters of “best method performance”


Step 5 – Numerical Solution Search, Best Conditions

Fusion MD identifies study parameters of “best method performance”


Graphical Tools Visualize factor effects with overlay graphs

Define performance goals (No. of peaks > 8)

Visualize effects

Shaded region indicates Final % and Gradient Time combinations that do not meet performance requirements.


Add effects to build the Design Space

2 performance goals

Define performance goals (No. of peaks > 8, RT < 3 min)

Visualize effects

Shaded regions indicate

Final % and Gradient Time

combinations that do not

meet performance

requirements.


Combine All Performance Goals Establish the Design Space

Design Space = Operating space Robustness

Mean Performance +


Next Phase Of Method Development

Option 1: Confirm optimised settings

– Run the sample with predicted settings

– Automatically built methods and sample set

– Method is ready to be validated

Option 2: Optimise these predicted settings

– Select best column, best solvent

– Fine tune secondary parameters

o Slope, flow rate, column temperature


Summary & Conclusions

FusionMD is a tool for developing LC methods according to

Quality-by-Design (QbD) guidelines

– Automates the entire process of method development

– Saves time by determining the minimum number of

experiments needed for valid results

– Provides tools to vizualise the impact of chromatographic

parameters

– Integrates robustness during development

FusionMD is a part of a System Solution

– Empower

– UPLC chemistry

– UPLC hardware


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Automated Method Development With UPLC and Fusion Software ... · Automated Method Development With UPLC and Fusion Software A Quality by Design-based Tool ©2012 Waters Corporation