VALIDATING CHROMATOGRAPHIC METHODS - idqc-hcm.gov.vn...VALIDATING CHROMATOGRAPHIC METHODS A Practical Guide DAVID M. BLIESNER JWUS_VC-Blies_FM.qxd 7/17/2006 4:02 PM Page iii

VALIDATING CHROMATOGRAPHIC METHODS

JWUS_VC-Blies_FM.qxd 7/17/2006 4:02 PM Page i

VALIDATINGCHROMATOGRAPHICMETHODSA Practical Guide

DAVID M. BLIESNER

JWUS_VC-Blies_FM.qxd 7/17/2006 4:02 PM Page iii

Copyright © 2006 by John Wiley & Sons, Inc. All rights reserved

Published by John Wiley & Sons, Inc., Hoboken, New Jersey

Published simultaneously in Canada

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Library of Congress Cataloging-in-Publication Data:

Bliesner, David M.

Validating chromatographic methods : a practical guide/by David

M. Bliesner.

p. cm.

Includes bibliographical reference and index.

ISBN-13: 978-0-471-74147-3

ISBN-10: 0-471-74147-7 (acid-free paper)

1. Chromatographic analysis ––Validity. 2. Science –– Methodology.

I. Title.

QD79. C4B57 2006

543� 8 –– dc22 2005036658

Printed in the United States of America

10 9 8 7 6 5 4 3 2 1

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v

CONTENTS

PREFACE vii

1. OVERVIEW OF METHODS VALIDATION 1

2. COMPONENTS OF METHODS VALIDATION 8

3. STEP 1: METHOD EVALUATION AND FURTHER DEVELOPMENT 15

4. STEP 2: FINAL METHOD DEVELOPMENT AND TRIAL METHODS VALIDATION 28

5. STEP 3: FORMAL METHODS VALIDATION AND REPORT GENERATION 41

6. STEP 4: FORMAL DATA REVIEW AND REPORT ISSUANCE 48

7. SUMMARY 55

APPENDICES

I. GLOSSARY OF METHODS VALIDATION TERMS 57

II. TEMPLATE FOR AN EXAMPLE METHODSVALIDATION STANDARD OPERATING PROCEDURE (SOP) 72

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III. TEMPLATE FOR AN EXAMPLE END-USERREQUIREMENTS QUESTIONNAIRE 154

IV. TEMPLATE FOR AN EXAMPLE METHOD REVIEW CHECKLIST 156

V. TEMPLATE FOR AN EXAMPLE STANDARD TEST METHOD 159

VI. TEMPLATE FOR AN EXAMPLE METHODS VALIDATION PROTOCOL 169

VII. TEMPLATE FOR AN EXAMPLE METHODS VALIDATION REPORT 218

REFERENCES 283

INDEX 285

vi CONTENTS

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vii

PREFACE

Delphi Analytical Services, Inc. has spent the past several years helping com-panies in the pharmaceutical industry improve their level of compliance withcurrent good manufacturing practices (CGMPs). This involvement hasincluded large and small companies who have already been subject to regu-latory action from the Food and Drug Administration (FDA) as well as com-panies who are taking preventative measures to avoid regulatory action. Aspart of this effort, a significant amount of time has been spent reviewing ana-lytical and bioanalytical methods and methods validation documentation.

Unfortunately, our experience leads us to conclude that despite improvedguidance from the FDA, leadership by the International Conference onHarmonization (ICH), and the plethora of validation experts and courses, asubstantial need still exists for education, training, and periodic retraining inthe field of methods validation.

Make no mistake, analytical methods validation is not a trivial undertak-ing. And like snowflakes, no two are exactly alike. However, our experiencehas brought us in contact with what we consider to be the best practices in theindustry. In addition, we have seen some of the mistakes that often degradethe overall quality of the finished product: A validated, transferable, analyti-cal method that will serve its end users for an extended period of time withminimal complications.

In our experience, very few labs have it “all together” and execute all thecomponents of a methods validation well. This guide was written with theintent to bring order to the potentially chaotic process of methods validation.

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If you are new to methods validation, we hope to provide you with enoughpractical information and tools to keep you from having to reinvent the wheelby having to develop your own systems and to attack methods validationfrom scratch. If you are experienced with methods validation, we hope youwill use this guide to upgrade and improve your existing systems.

This guide focuses on chromatographic methods validation, specificallyhigh performance liquid chromatographic (HPLC) methods validation. Thisapproach was chosen in that HPLC is by far the most common analyticaltechnique used in modern pharmaceutical analytical R&D/QC laboratories.However, the concepts are in many cases directly applicable to validation ofother analytical techniques as well.

In that CGMPs are always changing (hence the “C” meaning “current”)and the industry is always improving and upgrading its best practices, weencourage your feedback and comments so that we can keep this guide in linewith the best practices of the industry. We look forward to your input andhope this guide assists you in your continuing quest for quality.

DAVID M. BLIESNER

viii PREFACE

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1.1 WHAT IS METHODS VALIDATION?

The FDA in its most recent publication, Guidance for Industry on AnalyticalProcedures and Methods Validation, states:

Methods validation is the process of demonstrating that analytical proce-dures are suitable for their intended use. The methods validation processfor analytical procedures begins with the planned and systematic collectionby the applicant of the validation data to support analytical procedures. [1]

What the FDA does not say is that the actual validation component of themethods validation process should be the culmination of a well-organized,well-planned, and systematically executed process that includes methoddevelopment, prevalidation studies, and finally, methods validation itself.Gone are the days where one did methods development/validation concur-rently. Validation is the end game where few surprises and deviations areexpected. Validation is executed with a formal, approved and signed methodsvalidation protocol in place which has been reviewed by the quality assur-ance (QA) unit. Validation is complete when you:

(1) Demonstrate that you have met all the acceptance criteria.(2) Clearly document the results in a CGMP compliant fashion.

1

Validating Chromatographic Methods. By David M. BliesnerCopyright © 2006 John Wiley & Sons, Inc.

CHAPTER 1

OVERVIEW OF METHODS VALIDATION

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(3) Show how you met the acceptance criteria in a final methods valida-tion report, including references to raw data, all of which have beenreviewed and approved by the appropriate personnel including peers,management, and QA.

Some would even argue that the validation process is not complete untilthe methods are successfully transferred to their end-user laboratories.

This sounds like a daunting task. And to be completely honest—it is. Thereis nothing trivial or easy about methods validation. It takes time, resources,and rarely goes as easy as you think it’s going to go. Methods validation is partscience, part art, and a lot of bookkeeping and accounting. To be brutallyhonest, too few laboratories do a very good job executing all the components.

Due to the magnitude of the task, the time, and the perceived costs, manylaboratories try to cut corners. At a minimum, this results in deviationsfrom the protocol which no longer can be “arm waved” away. The FDAexpects you to scientifically address failures as you would any other labo-ratory investigation. This takes more time and effort and often results indelays in the validation timeline. In the worst case, you end up validating amethod that is transferred to quality control (QC) labs worldwide, and endsup being the root cause of untold laboratory investigations. It is hypothe-sized that many of the problems discovered during root-cause analysis ofout-of-specification results (OOS) are a direct result of poorly or partiallyvalidated methods.

The sections that follow provide a road map and the tools to guide andassist you to properly and efficiently validate your chromatographic methods,ensuring your validated methods do not become the root cause of your futurelaboratory investigations.

1.2 STEPS IN THE CHROMATOGRAPHICMETHODS VALIDATION PROCESS

The process of validating chromatographic methods can be broken down intofour steps. These steps include:

● 1. Method evaluation and further method development,● 2. Final method development and trial methods validation,● 3. Formal methods validation, and ● 4. Formal data review and report issuance.

Figure 1.1 graphically represents the process.

2 OVERVIEW OF METHODS VALIDATION


The estimates given in Figure 1.1 are only that—estimates. So manyvariables exist during methods validation that it makes it very difficult, if notimpossible, to give an accurate prediction of the length of the validation proj-ect. There is an enduring myth in the pharmaceutical industry that it shouldonly take six weeks to validate a method. The formal validation portion itselfshould only take about six weeks, but the preparation and documentation takesignificantly more time. Although it can be done, a complete validation,including proper data review and documentation, even for an established prod-uct with known properties, has not been satisfactorily completed in such ashort time. Industry professionals believe that this is why there are so manybad methods in use today.

Details of the design and implementation for each step are described in theremaining sections of this guide. In addition, many of the tools, templates,and examples needed to complete methods validation are included in the

STEPS IN THE CHROMATOGRAPHIC METHODS VALIDATION PROCESS 3

STEP 1Method Evaluation and Further Method Development

(~1 to 2 Months)

STEP 2Final Method Development and Trial Methods Validation

(~3 to 4 Months)

STEP 3Formal Methods Validation and Report Generation

(~1 to 2 Months)

STEP 4Formal Data Review and Report Issuance

(~1 to 2 Months)

FIGURE 1.1 Steps in the chromatographic methods validation process. Total time formethod evaluation, validation, data/documentation review, and reporting is approximately 6 to10 months.


appendices. As many examples, based on real-world scenarios, as practicalhave been provided to give you a framework to validate your own methods.In particular, the following key elements which constitute a methods valida-tion program have been provided:

● A template for a methods validation standard operation procedure (SOP)example

● A template for a standard test method example● A template for a methods validation protocol example● A template for a methods validation report example

Each template for these examples represents a significant body of knowl-edge and experience. It would require a substantial amount of your time tocreate these templates independently. Modify them and use them to best fitthe practices within your organization.

1.3 HOW TO USE THIS GUIDE

Before proceeding, here are some suggestions on how to use this guide.Although this document is a practical guide, it is by no means a technicalcookbook on how to validate an analytical method. This would be impossi-ble since every method has its own unique idiosyncrasies. Therefore, it isrecommended that you take the following approach to best apply thisguide:

● Familiarize yourself with the standard methods validation terms listed inthe glossary.

● Read and understand the guide section titled “Components of a methodsvalidation.”

● Review the template for the methods validation SOP example.● Review the template for the test method example.● Review the template for the methods validation protocol example.● Read the template for the methods validation report example.● Read and understand the flowcharts and checklist related to methods val-

idation in steps 1– 4.● Develop your own systems and templates by adapting the systems and

templates presented in this guide to your laboratory as appropriate.● Train your chemists on the systems.● Implement your systems.



1.4 ADDITIONAL POINTS TO CONSIDER WHEN VALIDATINGCHROMATOGRAPHIC METHODS

In addition to the nuts and bolts of methods validation, many soft skills existthat will improve your chances of success during validation. Therefore, asyou apply this guide to methods validation in your own laboratory, pleasekeep the following points in mind.

Do Not Underestimate the Value of Planning and Organization

Much of methods validation is bookkeeping—both figuratively and literally.Therefore, much of the success of methods validation is dependent upon theamount of effort and attention to detail made in steps 1 and 2. The frameworkof what constitutes a methods validation is predetermined by FDA, ICH, andcurrent industry practice. Think systematically and work with the end in mind.Therefore, by prioritizing and planning your work carefully, allocating yourresources efficiently, coupled with good supervision and communications,you will significantly enhance your chances of a positive and timely outcome.

Make It Simple, Keep It Simple and Remember Your End Users

Chromatographic methods are often developed and validated by analyticalresearch-and-development scientists who are not the end users of the method.Because of this, end user requirements are often not taken into consideration,which may lead to an overly complex and scientifically elegant method thatwill be “thrown over the wall” to the quality control (QC) chemists duringtechnology transfer. Despite the myth, QC chemists and technicians are usu-ally technically sound and well educated. However, they often work in a pres-sure cooker environment where the complexity and nuances of a method willonly make their life more difficult. They need to get product out the door,with minimal complications and effort. Therefore, methods should be madeas simple and robust as possible, with the end user’s needs in mind.

QA Is Your Friend

It was once said the best quality assurance (QA) person is the one who eats hisor her lunch alone because none of the chemists wants to talk with him or her.Unfortunately there is often an unhealthy tension that develops between QAand the lab. You should make every effort to reduce such tensions during themethods validation process. Remember, in the future your work will bereviewed cold, without coaching from you, by an FDA reviewer. Therefore,you need to develop and present a complete and accurate account of your work

ADDITIONAL POINTS TO CONSIDER WHEN VALIDATING CHROMATOGRAPHIC METHODS 5


which raises few questions. So look at QA as your first line of defense. If youcan’t make them understand your work, how will the FDA understand it?

Don’t Underestimate the Value of Experience

As stated, no two methods validations are the same, but each is very similar.This means that someone somewhere has probably encountered the sameproblems you are encountering. As part of your step 1 planning or trou-bleshooting, consider who might have experience with your work or workthat is similar to what you are doing. Look within your own organization atprevious validations. Look to vendors outside your organization such as theAPI manufacturer, your reagent supplier, and your chromatographic equip-ment and material supplier. Go to the library, search the literature, and tapinto your network within the industry. Chances are someone has an answer oreven the answer to your question. Avoid the “not invented here syndrome.”Don’t fall in love with your own work and skills to the exclusion of othergood ideas. Ph.D. level R&D scientists are particularly bad about this.Remember the goal and resist the temptation of creating another dissertationresearch project.

Common Sense Is an Uncommon Virtue

During the course of the validation process perform what we call periodicsanity checks. Stop and ask yourself: Do these results make sense? Does thissolution to the problem make sense for my end users? Am I headed in theright direction? Don’t be shy about talking to your end users as well.

Mistakes Are Made Under Pressure

The validation of bad methods invariably comes by having to perform thevalidation under pressure. Again, this is why it’s so important to expend a sig-nificant effort on steps 1 and 2 of the validation process. When you get tomonth six and still don’t have a functional (let alone validated) method, pres-sure will make you get the methods out to your end users when they nevershould have made it out of your lab. People make mistakes, but people makemore significant mistakes under pressure.

Realize the Impact of Your Successes and Your Failures

The lifetime of a validated chromatographic method can span decades. Themethod may be used in laboratories all over the world. The financial andresource usage impact of the method can be substantial. Keep this in mind as



you validate your methods. The rule is once the method is validated andtransferred, the chances of changing anything significantly with the methodare very limited. From a regulatory standpoint, this situation raises a signifi-cant number of questions. From a practical standpoint, it costs too much andtakes away resources from the next project. Be mindful of these points and doit right the first time. Remember:

Validating an analytical method may be the most important task youwill perform during your tenure with your company.

ADDITIONAL POINTS TO CONSIDER WHEN VALIDATING CHROMATOGRAPHIC METHODS 7


2.1 BACKGROUND

Chromatographic methods validation is subdivided into four categories,generally recognized via the United States Pharmacopoeia (USP). Thesecategories include:

Category I. Validation of analytical methods for assayCategory II. Validation of analytical methods for impurities and

degradantsCategory III. Validation of analytical methods for dissolutionCategory IV. Validation of analytical methods for identification

Although clearly separated, validation can encompass more than onecategory simultaneously, depending on the workplan or resources available.For example, a method may be (and frequently is) validated concurrently forassay and for impurities and degradants. This is true because many of thevalidation characteristics tested during assay validations are the same as forimpurities and degradants. A description of the validation characteristics arelisted in the following sections.

CHAPTER 2

COMPONENTS OF METHODSVALIDATION

8


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2.2 APPROACH

The following are typical analytical performance characteristics which maybe tested during methods validation:

● Accuracy● Precision

� Repeatability� Intermediate precision

● Specificity● Detection limit● Quantitation limit● Linearity● Range● Robustness● System suitability determination● Forced degradation studies● Solution stability studies● Filter retention studies● Extraction efficiency studies● Additional methods validation information

� Representative instrument output� Representative calculations� Listing and characterization of known impurities� Degradation pathways (if known)

A more detailed definition of each characteristic is given in the followingsubsections.

2.2.1 Accuracy

Accuracy is the nearness of a measured value to the true or accepted value. Itprovides an indication of any systematic error or bias in the method. For anunbiased method, a theoretical plot of measured value versus true value canbe described by a mathematical function. In the pharmaceutical industry, thisis typically a straight line with a given slope and zero intercept. It follows thatthe accuracy of a biased method varies with the analyte concentration accord-ing to the types of systematic errors.

During the validation, accuracy is determined by measuring the recoveryof the active component from a drug product matrix or by directly measuring

APPROACH 9


the active pharmaceutical ingredient (API). Typically studies involve spikingthe drug product placebo matrix with API in amounts equal to the nominalfinished dosage strength. This spiking is either by adding of standardsolutions or dry spiking API into the matrix followed by complete mixing.

2.2.2 Precision

Precision consists of two components: repeatability and intermediate preci-sion. Repeatability is the variation experienced by a single analyst on a singleinstrument. Repeatability does not distinguish between variation from theinstrument or system alone and from the sample preparation process. Duringthe validation, repeatability is performed by analyzing multiple replicates ofan assay composite sample by using the analytical method. The recoveryvalue is calculated and reported for each value.

Intermediate precision refers to variations within a laboratory such as dif-ferent days, with different instruments, and by different analysts. This wasformerly known as ruggedness. During the validation, a second analystrepeats the repeatability analysis on a different day using different conditionsand different instruments. Recovery values are calculated and reported. A sta-tistical comparison is made to the first analyst’s results.

2.2.3 Specificity

Specificity is the ability to assess unequivocally the analyte in the presenceof components that may be expected to be present such as impurities,degradation products, and excipients. There must be inarguable data for amethod to be specific. Specificity � measures only the desired componentwithout interference from other species that might be present; separation isnot necessarily required.

To determine specificity during the validation blanks, sample matrix(placebo), and known related impurities are analyzed to determine whetherinterferences occur. Specificity is also demonstrated during forced degrada-tion studies.

The term “selectivity” is sometimes used interchangeably with specificity.Technically, however, there is a difference. Selectivity is defined as the abilityof the method to separate the analyte from other components that may bepresent in the sample, including impurities. Selectivity is separate and showsevery component in the sample. Therefore, one could have a method that isspecific, yet it may not be selective. For instance, an ion selective electrodemay be specific (e.g., is used to measure a single species in sample matrix),yet not be selective (e.g., doesn’t separate and identify all componentspresent).

10 COMPONENTS OF METHODS VALIDATION


2.2.4 Detection Limit

The detection limit (DL) or limit of detection (LOD) of an individualprocedure is the lowest amount of analyte in a sample that can be detected butnot necessarily quantitated as an exact value. The LOD is a parameter of limittests (i.e., tests that only determine if the analyte concentration is above orbelow a specification limit).

In analytical procedures such as HPLC that exhibit baseline noise, the LODcan be based on a signal-to-noise (S/N) ratio (3:1), which is usually expressed asthe concentration (e.g., percentage, parts per billion) of analyte in the sample.There are several ways in which it can be determined, but it usually involvesinjecting samples, which generate an S/N of 3:1, and estimating the DL.

2.2.5 Quantitation Limit

The quantitation limit (QL) or limit of quantitation (LOQ) of an individualanalytical procedure is the lowest amount of analyte in a sample that can bequantitatively determined with suitable precision and accuracy. The quantita-tion limit is a parameter of quantitative assays for low concentrations ofcompounds in sample matrices and is used particularly for the determinationof impurities and/or degradation products. It is usually expressed as the con-centration (e.g., percentage, parts per million) of analyte in the sample.

For analytical procedures such as HPLC that exhibit baseline noise, theLOQ is generally estimated from a determination of S/N ratio (10:1) and isusually confirmed by injecting standards which give this S/N ratio and havean acceptable percent relative standard deviations (%RSDs) as well.

2.2.6 Linearity

Linearity evaluates the analytical procedure’s ability (within a give range) toobtain a response that is directly proportional to the concentration (amount) ofanalyte in the sample. If the method is linear, the test results are directly or bywell-defined mathematical transformation proportional to the concentration ofanalyte in samples within a given range. Linearity is usually expressed as theconfidence limit around the slope of the regression line. The line is calculatedaccording to an established mathematical relationship from the test responseobtained by the analysis of samples with varying concentrations of analyte.Note that this is different from range (sometimes referred to as linearity ofmethod), which is evaluated using samples and must encompass the specifica-tion range of the component assayed in the drug product.

During validation, linearity may be established for all active substances,preservatives, and expected impurities. Evaluation is usually performed onstandards.

APPROACH 11


2.2.7 Range

Range is defined as the interval between the upper and lower concentrations(amounts) of analyte in the sample (including these concentrations) for whichit has been demonstrated that the analytical procedure has a suitable level ofprecision, accuracy, and linearity.

Range is normally expressed in the same units as test results (e.g., percent,parts per million) obtained by the analytical method.

During validation, range (sometimes referred to as linearity of method) isevaluated using samples (usually spiked placebos) and must encompass thespecification range of the component assayed in the drug product.

2.2.8 Robustness

Robustness is defined as the measure of the ability of an analytical method toremain unaffected by small but deliberate variations in method parameters (e.g.,pH, mobile-phase composition, temperature, and instrument settings) and pro-vides an indication of its reliability during normal usage. This is an importantparameter with respect to the transferability of the method following validation.

Determining robustness is a systematic process of varying a parameter andmeasuring the effect on the method by monitoring system suitability and/orthe analysis of samples. It is part of the formal methods validation process.

2.2.9 System Suitability Determination

System suitability is the evaluation of the components of an analytical systemto show that the performance of a system meets the standards required by amethod. A system suitability evaluation usually contains its own set ofparameters. For chromatographic assays, these may include tailing factors,resolution, and precision of standard peak areas, and comparison to a confir-mation standard, capacity factors, retention times, and theoretical plates.

During validation, where applicable, system suitability parameters arecalculated, recorded, and trended throughout the course of the validation.Final values are then determined from this history.

2.2.10 Forced Degradation Studies

Forced degradation or stress studies are undertaken to deliberately degradethe sample (e.g., drug product, excipients, or API). These studies are used toevaluate an analytical method’s ability to measure an active ingredient andits degradation products, without interference, by generating potentialdegradation products.



During validation, samples of drug product (spiked placebos) and drugsubstance are exposed to heat, light, acid, base, and oxidizing agent to pro-duce approximately 10% to 30% degradation of the active substance. Thedegraded samples are then analyzed using the method to determine if thereare interferences with the active or related compound peaks. Forced degrada-tion studies can be time consuming and difficult because it is often difficultto generate the proper level of degradation. Also, a certain amount of logicneeds to be applied to extrapolate the results of these studies to what might beseen during actual stability studies.

2.2.11 Solution Stability Studies

During validation the stability of standards and samples is established undernormal benchtop conditions, normal storage conditions, and sometimes inthe instrument (e.g., an HPLC autosampler) to determine if special storageconditions are necessary, for instance, refrigeration or protection from light.Stability is determined by comparing the response and impurity profile fromaged standards or samples to the response and impurity profile of freshly pre-pared standards.

2.2.12 Filter Retention Studies

Filter retention studies are a comparison of filtered to unfiltered solutionsduring a methods validation to determine whether the filter being used retainsany active compounds or contributes unknown compounds to the analysis.Blank, sample, and standard solutions are analyzed with and without filtra-tion. Comparisons are made in recovery and appearance of chromatograms.

2.2.13 Extraction Efficiency Studies

Extraction efficiency is the measure of the effectiveness of extraction of thedrug substance from the sample matrix. Studies are conducted during meth-ods validation to determine that the sample preparation scheme is sufficientto ensure complete extraction without being unnecessarily excessive.Extraction efficiency is normally investigated by varying the shaking or son-ication times (and/or temperature) as appropriate during sample preparationon manufactured (actual) drug product dosage forms.

2.2.14 Additional Methods Validation Information Often Required

In addition to these analytical performance characteristics, the followinginformation is usually obtained or presented in the final validation report:

APPROACH 13


● Representative instrument output● Representative calculations● Listing and characterization of known impurities● Degradation pathways (if known)● Determination of relative response factor

Analytical methods performance characteristics are only part of the vocabu-lary that constitutes the language of methods validation. If you are new to themethods validation process, please review Appendix I, “Glossary of MethodsValidation Terms,” before proceeding.

2.3 THE BEST PLACE TO START

You must consider the use of the method and what validation characteristicsare crucial for you to determine the proper use of the method. For example, ifyou are validating a stability indicating method for related compounds, thenthe following order is probably best:

1. Selectivity 2. LOD/LOQ3. Forced degradation studies

If you have problems with selectivity, then you are dead before you start. Youneed to know this at the beginning. If you cannot detect a related compoundsat the lowest level needed, then you won’t be able to ensure you are meetingyour specifications.

Perform a sanity check at the beginning so you don’t have an “oops”halfway through your validation.



3.1 BACKGROUND

Although it has been stated that the FDA expects methods validation to be anindependent and separate process from method development, it is sometimesvery difficult for a laboratory to distinguish between the two functions. Thepurpose of step 1: Method evaluation and further method development is toassist in organizing your thoughts and guiding you in making the transitionfrom method development to methods validation.

In some circumstances this transition will be straightforward and simple.For example, if a significant amount of research and development has beenperformed and a development report generated, step 1 will be a sanity checkto make sure all the proper effort has been completed prior to prevalidationand formal validation studies (e.g., limited evaluation and no additionaldevelopment are needed). However, if you have just been handed an olderexisting method that has not been validated to existing standards, step 1 needsto be executed with greater care and attention to detail (e.g., significant eval-uation and perhaps substantially additional development are needed).

Regardless, we assume that by the time you are ready to begin step 1 youhave performed a certain amount of method development yourself or have

15


CHAPTER 3

STEP 1: METHOD EVALUATION ANDFURTHER DEVELOPMENT

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found a separation in the literature or from another source which fits yourneeds.

Step 1 does have a clear deliverable objective: To identify a method suit-able for the end user’s intended needs and stands a reasonable chance ofbeing validated without complications. These criteria must be supported bydata. Remember, the more initial effort, the better the entire validation willproceed and the higher the quality of the finished product.

It should be noted that in this guide, we chose not to spend significant timeon the concept (and art) of method development. This is an entire topic untoitself and more of a scientific endeavor than an act of regulatory compliance.We simply offer a means to gather and organize information which can beused to develop or evaluate your method and make the transition from devel-opment to validation.

3.2 APPROACH

The basic elements of step 1 are:

● Methods validation commissioned in writing● Methods validation team leader assigned● Background information and end user requirements sourced, collected,

and compiled ● Validation team leader selects validation team members● Background information distributed for review and evaluation by the

validation team● Planning meetings held; work breakdown structure and project plan

created● Team member training conducted● Project plan executed● Development review report/summary report generated

Regardless of how you proceed, however, much of your validation effortsshould be guided by your company’s methods validation standard operatingprocedure (SOP). A template of a methods validation SOP example ispresented in Appendix II and should be reviewed at this point if you are newto the methods validation process. Steps in this process are shown in Figure3.1 and described in Table 3.1.

16 STEP 1: METHOD EVALUATION AND FURTHER DEVELOPMENT


APPROACH 17

Components of Step 1:Method Evaluation and Further Method Development

~1 to 2 Months

Methods ValidationCommissioned in Writing

Step 1.1 0.25 Days

Initiate Methods ValidationProcess (Step 1: MethodEvaluation and Further

Development)

Step 1.2 0.25 Days

Methods Validation TeamLeader Assigned

Step 1.3 0.25 Days

Team Leader Obtains allAppropriate Existing

Documentation Related toMethods Validation andSpecific Method to be

Validated

Step 1.4 5.0 Days

Team Leader ContactsPersonnel Associated with

Previous and/or SimilarValidations/Development to

Solicit Information andAdvice

Step 1.5 2.0 Days

Team Leader ReviewsDocumentation and Gleans

Information of Value/Necessity for Current

Validation

Step 1.6 2.0 Days

FIGURE 3.1 Diagram of workflow for step 1: method evaluation and further methoddevelopment.


18 STEP 1: METHODS EVALUATION AND FURTHER DEVELOPMENT

Team Leader CompletesEnd User Requirements

Analysis IncludingContacting End User Labsand Obtaining Additional

Clarification onCommissioning Document

Step 1.7 2.0 Days

Team Leader SelectsValidation Team Members

Step 1.8 0.50 Days

Team Leader Compiles andDistributes DocumentationPackage for Team Member

Review

Step 1.9 0.50 Days

Team Leader SchedulesPlanning Meeting with

Team Members

Step 1.10 0.50 Days

Team Members ReviewDocumentation and Prepare

for Planning Meeting

Step 1.11 2.0 Days

Team Leader CreatesAgenda and Presentation

for Planning Meeting

Step 1.12 0.25 Days

FIGURE 3.1 (Continued)


APPROACH 19

Planning Meeting Held withValidation Team Members

Step 1.13 0.50 Days

Team Leader Creates "Step 1: Method Evaluation and

Further MethodDevelopment Project Plan"

From Work BreakdownStructure

Step 1.14 1.0 Days

Team Leader PromulgatesProject Plan for Step1 andSolicits Team Feedback

Step 1.15 2.0 Days

Team Members ApproveProject Plan

Step 1.17 2.0 Days

Team Leader CreatesTraining Plan

Step 1.18 2.0 Days

No

Changes toProject Plan?

Step 1.16

Team Leader MakesChanges

Step 1.16.1 0.50 Days

Yes




Execute and DocumentTraining

Step 1.19 2.00 Days

Review Step 1 Project Planand Timeline With Team

Members

Step 1.20 0.25 Days

Execute Step1 Project Plan

Step 1.21 10.0 Days

Team Leader Collects,Compiles and Reviews

Results

Step 1.22 2.0 Days

Summarize Results in aDevelopment Review

Report/Summary

Step 1.23 2.0 Days

Team Leader CirculatesDevelopment ReviewReport/Summary to

Validation Team for Reviewand Input

Step 1.24 2.0 Days

Proceed to Step 2:Final Method Development and Trial Methods Validation



TABLE 3.1 Explanation of Workflow Diagram Steps for Step 1:Method Evaluation and Further Method Development

EstimatedStep Description Duration Explanation

1.1 Methods validation 0.25 day The success of any project, including methodcommissioned in validation, depends on managementwriting commitment and involvement. Therefore,

it is important that management, within theorganization responsible for methodsvalidation, formally commission the validationin writing. As part of this commissioning,management should: (1) assign a team leader,(2) define the product type which will requirethe analytical methodology, (3) define theUSP method category under which themethod will be validated, (4) provide requiredstart and end dates, (5) define method transfergoals and dates of transfer, and (6) determinewhether the validation will be full, partial, orcompendial.

1.2 Initiate methods 0.25 day This should correspond to the start datevalidation process determined by management and cited in the (step 1: method evaluation commissioning document. and further method development)

1.3 Methods validation 0.25 day As described in step 1.1, assignment of theteam leader assigned team leader should be part of the methods

validation commissioning procedure. Theselection of a methods validation team leaderis important. The team leader will be heldaccountable for the successful completion ofthe validation within the expected timeframe.An individual with the appropriate technicalskills as well as project management andorganizational skills is required. A goodunderstanding of laboratory CGMPs andCGMP documentation requirements is alsoimportant.

1.4 Team leader obtains 5.0 days The following documentation should be all appropriate existing sourced if available and/or appropriate: (1) documentation related method development SOP, (2) method to methods validation validation SOP, (3) method development and specific method reports, (4) API manufacturer technicalto be validated reports and documents, (5) existing written

methods, (6) USP, BP, and EP methods,

APPROACH 21

(Continued)


(7) existing methods validation packages forthis product or chromatographic separation,(8) existing methods validation packages forsimilar products or molecules, (9) ICHguidelines and FDA guidance documents,(10) selected text references such asAnalytical Profiles of Drug Substances andExcipients, [3], (11) scientific literaturesearch (paper or electronic) results for theproduct, drug substance, or relatedcompounds, (12) queries of HPLC columnvendors, HPLC supply vendors, etc., (13)other sources of information which may havephysical-chemical properties of active andexcipients including solubility, pKa, spectralproperties, etc. (Note: See Appendix II for an exampletemplate of a methods validation SOP.)

1.5 Team leader contacts 2.0 days As indicated in step 1.4, it is important to personnel associated investigate if methods have been developed with previous and/or and/or validated previously for your similar validations/ molecules and products. Often work has development to solicit already taken place within your company or information and advice another company. Do not pass up the

opportunity to talk with the people who havealready performed the work. In many cases,even if they are not from your company, theywill be willing to share their experiences. Inturn, they may be able to save you a lot of time.

1.6 Team leader reviews 2.0 days Once the information and feedback is information of obtained, the team leader gleans the pertinent value/necessity for information and compiles a concise current validation documentation package for later distribution

to the validation team.

1.7 Team leader completes 2.0 days As part of the preparation process for end-user requirements methods validation, it is imperative to obtain analysis including feedback from the individuals who will be contacting end-user the end users of the method. Some of the labs and obtaining questions which should be asked include: (1)additional clarification Who will use the method (e.g., QC lab for on commissioning routine testing, in-process lab)? (2) Whatdocument types of equipment are available and are

there any equipment limitations? (3) Where


TABLE 3.1 (Continued)



will the method be used (e.g., geographiclocations and actual facility location)? (4)What supply restrictions may exist? (5) Whatexpertise (e.g., the education, training, andexperience levels of the end users)restrictions may exist? (6) What languagebarriers may exist? (7) Under whichUSP/ICH methods category does the methodfall? (8) What validation characteristics arerequired? (Note: See Appendix III for anexample of a template of an end-userquestionnaire.)

1.8 Team leader selects 0.5 day Some selection criteria for team membersvalidation team should include the following: (1) Is there a members skill match to similar product/molecules and

techniques? (2) Is there an experience matchto similar products/molecules andtechniques? (3) Are the personnel currentlyavailable to work on the project and can theyhandle the additional workload? (3) What isthe employment status and person situationof the potential team members (e.g., theymay be in line for a promotion which wouldtake them away from the project)? (4) Dothey have the ability to work closely with theother team members?The team member selection should bereviewed, approved, and then communicatedthrough management. This will ensureconflicts no with personnel, scheduling, orresearch allocation.

1.9 Team leader compiles 0.5 day This documentation package should include:and distributes (1) the commissioning document, (2) the documentation package current revision of the methods validation for team member review SOP, (3) existing development reports and

documentation, (4) existing methods, and (5)the end-user requirements assessment.

1.10 Team leader schedules 0.5 day The team leader should make an attempt to planning meeting dovetail this meeting with the team members' with team members existing schedules.

APPROACH 23



(Continued)


1.11 Team members review 2.0 days Team members should perform a thorough documentation and review of the package prior to attending the prepare for planning meeting. They should then generate a list of meeting questions to present at the first team meeting.

1.12 Team leader creates 0.25 day The agenda should include: (1) team member agenda and presentation introductions, (2) team member feedback on for planning meeting the end-user requirement responses, (3)

timeline analysis, (4) quality of existing dataand documentation, (5) team memberexperience with the product, molecules, orproject, (6) team member trainingrequirements, (7) team member workpreferences (e.g., preferences for executingcertain portions of the validation or use ofspecific analytical techniques), (8) a groupbrainstorming session to identify anypotential difficulties prior to creating aproject plan.

1.13 Planning meeting held 0.5 day The purpose of the meeting is threefold: (1) with validation team to formally establish the team, (2) to inform members: Work team members on the details of the project,breakdown structure and (3) to have the team leader obtain is created feedback from the team members so the team

leader can develop an accurate and realisticproject plan and make appropriate workassignments.

1.14 Team leader creates 1.0 day The project plan should be a straightforward step 1: method document, such as a calendar, which evaluation and further delineates the tasks required to evaluate the method development method and/or the tasks required to perform project plan from work additional methods validation. Assignment of breakdown structure team member responsibilities and start and

stop dates are also included in the projectplan. Although each project plan will beunique, some example tasks which may beincluded in the project plan include:

An example “best case” scenario (e.g.,method previously developed, completemethod development report and packageexists):

(1) Evaluate development anddocumentation for API standard, relatedcompound standards, placebo, and spiked





placebo with existing method to confirmthat separation is valid.

(2) Evaluate chromatography presented indevelopment documentation for upperand low ends of desired linear range.

(3) Perform a sanity check on the separationwith respect to compound solubility,stability of solutions, pKa versus pH ofthe buffer, toxicity of materials,appropriateness of lambda max, etc.

(4) Review end-user’s requirements andmatch to method characteristics to ensuremethod will work for intended use.

(5) Perform confirmatory separation withmethod and calculate fundamentalchromatographic figures of merit,including LOD/LOQ if appropriate forAPI standard, related compoundsstandards, placebo, and spiked placebo asappropriate.

An example “worst case” scenario (e.g.,literature references for methods only):

(1) Perform a sanity check of literatureseparation with respect to compoundsolubility, stability of solutions, pKaversus buffer pH, toxicity of materials,appropriateness of lambda max, etc.

Note: See Appendix IV for an example ofa template of a method review checklistfor summary sanity check question to beconsidered.

(2) Perform a limited robustness study toinclude mobile phase adjustments, pHadjustments, temperature changes,change of column manufacturer, etc.

(3) Perform LOD and LOQ tests asappropriate.

(4) Review end-user requirements and matchto method characteristics to ensuremethod will work for intended use.

(5) Determine “go” or “no go” on themethod as written.

APPROACH 25



(Continued)


Note: At this stage, the team may have asubstantial amount of information aboutthe separation, or perhaps very littleinformation. It depends entirely uponhow much development work has beenperformed prior to this point.

Note: This project plan is specificallydesigned for step 1 only.

1.15 Team leader promulgates 2.0 days This gives the team members the opportunity project plan for step 1and to make corrections or modifications to the solicits team feedback project plan for this phase of the validation.

1.16 Changes to project 0.5 day If team members have made suggestions for plan? If yes, team changes, the team leader will make the leader makes changes changes as appropriate and recirculate the and forwards back plan for additional feedback. Management to team members should also be given the opportunity to for feedback review the draft project plan.

1.17 Team members 2.0 days Once all the changes have been made, the approve project plan team should come to an agreement on the

plan's content and move forward towardimplementation. This acceptance should beshared with all team members andmanagement.

1.18 Team leader creates 2.0 days Once the project plan is accepted, training of training plan the team members needs to be executed.

Some areas which may require traininginclude: (1) SOP training/retraining, (2)analytical technique training/retraining, (3)training on methods validation protocolexecution, (4) data capture and reviewprocedures, (5) workflow procedure review,(6) data reporting procedure review, and (7)laboratory investigation reporting (LIR)procedures required during validation.

Although these topics may not be applicableto the method evaluation and furtherdevelopment phase, they will be applicableduring formal methods validation. Additionaltopics may be included during this training.





1.19 Execute and document 2.0 days Self explanatory training

1.20 Review step 1 0.25 day Following training, and prior to executing theproject plan and timeline project plan, the plan and timeline should be with team members reviewed with all team members one last time.

1.21 Execute step 1 10.0 days Start date should be formally communicated project plan to management and all team members.

1.22 Team leader collects, 2.0 days As data are obtained or an analysis of compiles, and existing data is conducted by the teamreviews results members during methods evaluation, the

team leader begins compiling and organizingthe findings to be included into adevelopment review/summary report.

1.23 Summarize results 2.0 days At this point, the team leader will generate ain a development development review and similar summary review report/summary report.

1.24 Team leader circulates 2.0 days The development review or summary report development review is circulated among the team members for report/summary to review and input and in preparation for a validation team for group review session. review and input

Proceed to step 2: final method developmentand trial methods validation.

APPROACH 27




28

4.1 BACKGROUND

At the conclusion of the tasks delineated in step 1, many organizations launchimmediately into formal methods validation. In many cases this is a riskyapproach. Although the data may support the fact that the method will workfor its intended use, the components of methods validation make it a veryrigorous process, fully testing the ability for the method to be used consis-tently in many different laboratory environments.

Recall that the FDA expects validation to be the end game, where few sur-prises and deviations are encountered. Moreover, methods validation must beinitiated and executed by creation and implementation of a methods valida-tion protocol. If deviations or modifications from the protocol occur duringvalidation, documentation is required to explain the deviations and supportthem with data. You cannot simply explain away failures or modify the pro-tocol in order to make the experiments successful.

In many cases, failure to meet protocol acceptance criteria often leads toadditional methods development. This in turn requires modification of the exist-ing protocol or even creation of a new protocol with subsequent revalidation.Obviously this do-over cycle can take a tremendous amount of additional time


CHAPTER 4

STEP 2: FINAL METHODDEVELOPMENT AND TRIAL METHODS VALIDATION


and effort. Therefore, it is in the laboratory’s best interest to make sure themethod works as intended before executing the methods validation protocol.This leads us to step 2: final method development and trial methods validation.

The purpose of step 2 is to use the data obtained in step 1, to make a logi-cal determination that the method can be validated, and then to create a trialor practice methods validation protocol and implement a trial methodsvalidation via the protocol.

Since the trial protocol is not an official document and the data collected willnot be included in any type of regulatory submission, the laboratory is free tomodify the experiments and alter acceptance criteria as necessary. Although thismay seem like overkill, experience shows that invariably difficulties are encoun-tered during the first pass of a validation, and because the data are now part of aregulatory submission, the time and effort it takes to implement the do-overcycle easily surpass the time and effort required to perform a trial validation.

The clear deliverables for step 2: final method development and trialmethods validation include:

1. A final methods validation protocol with established, reasonable, andobtainable acceptance criteria

2. A method which you know you can validate

If you have some previous experience conducting method developmentand/or performing methods validation, this approach may seem like overkill.However, experience shows that difficulties are invariably encounteredduring the first pass of a validation. Because of this, the value of a trial meth-ods validation cannot be overemphasized. Therefore, the best guidance wecan give to you is:

Do not gamble! Perform a trial methods validation first.

Regardless of your decision, templates of an example methods validationprotocol and a standard test method are included for your review inAppendices VI and V, respectively.

4.2 APPROACH

The basic elements of step 2 are

● Validation team discusses development reports or summaries to assesswhether further development work is needed, and to initiate work asappropriate

● Create trial methods validation protocol

APPROACH 29


● Create methods validation project plan● Implement trial methods validation via protocol and project plan● Collect and review data ● Create trial methods validation report ● Compare results obtained during trial validation versus the trial protocol● Determine if method is validatable; modify experiments and protocol as

necessary● Execute additional experiments as necessary● Proceed to formal methods validation

Steps in this process are shown in Figure 4.1 and described in Table 4.1.

30 STEP 2: FINAL METHOD DEVELOPMENT AND TRIAL METHODS VALIDATION


APPROACH 31

FIGURE 4.1 Work flow diagram for step 2: final method development and trial methodsvalidation.

Validation TeamDiscusses

DevelopmentReview Report/Summary as aGroup in Open

ForumStep 2.1 1.0 Days

Is Method Ready for TrialMethods Validation?Step 2.2 0.25 Days

No

Team LeaderCreates/

Completes DraftMethod

Step 2.3 1.00Days

Validation TeamIdentifies

Weaknesses inExisting MethodStep 2.2.1 0.25

Days

Team LeaderCreates Draft Trial

MethodsValidation Protocol

Step 2.5 2.00Days

Team LeaderCirculates Draft

Method toAppropriate Team

Members forReview and Input

Step 2.4 2.00Days

Validation TeamOutlines

[Additional]Development PlanStep 2.2.2 0.50

Days

Team LeaderCreates

Development PlanStep 2.2.3 1.00

Days

Development PlanCirculated to Team

for AgreementStep 2.2.4 2.00

Days

Development PlanIssued andExecuted

Step 2.2.5 20.0Days

Team LeaderCompiles Results

in a ReviewReport/SummaryStep 2.2.6 2.00

Days

Components of Step 2:Final Method Development and Trial Methods Validation

~3 to 4 Months

Yes



Team MembersConcur on DraftMethod ContentStep 2.6 1.00

Days

Team MembersConcur on DraftTrial Methods

Validation ProtocolStep 2.7 1.00

Days

Team LeaderCreates WorkBreakdownStructure

Step 2.8. 1.00Days

Team LeaderPromulgates

Project Plan toTeam MembersStep 2.10 1.00

Days

Changes toProject Plan?

Step 2.11 0.00Days

Team MembersApprove Project

PlanStep 2.12 0.50

Days

Team LeaderMakes ChangesStep 2.11.10.50

Days

Team LeaderCreates Project

PlanStep 2.9. 1.00

Days

Team LeaderSolicits FeedbackFrom QA on Draft

Trial MethodsValidation Protocol

Step 2.13 1.00Days


is FinalizedStep 2.16 1.00

Days


is Issued andExecuted

Step 2.18 0.25Days

Draft MethodFinalized as

Working MethodStep 2.14 1.00

Days

Draft MethodIncluded in Trial


Days

Yes

No

Team LeaderPerforms Reviewof Trial Methods

Validation Protocoland ValidationSOP to Insure

AlignmentStep 2.17 0.25

Days



APPROACH 33

Trial MethodsValidation ProjectPlan is ExecutedStep 2.19 0.25

Days

Trial MethodsValidation DataCollected andReviewed asAppropriate

Step 2.20 15.00Days

Team LeaderCompiles Trial

MethodsValidation Data in

Draft ReportFormat

Step 2.21 2.00Days

Compare ResultsObtained to Trial

Validation ProtocolAcceptance

Criteria RangesStep 2.22 0.50

Days

Team LeaderPromulgates Draft

Trial MethodsValidation Reportto Team Members

for Review andComment

Step 2.23 2.00Days

Team LeaderSolicits FeedbackFrom QA on Trial

MethodsValidation Report

Format andContent

Step 2.24 2.00Days

Trial ValidationReport Review

Meeting Held withValidation Team

Members and QAStep 2.26 0.50

Days

Team LeaderSchedules ReportReview Meeting

with TeamMembers and QAStep 2.25 0.50

Days

Is the Method “Validatable” viaExisting Method and Protocol?

Step 2.27 0.25 Days

Will modificationsRequire FurtherExperiments?

Step 2.28 0.25Days

Yes: Proceed to Step 3

Team LeaderMake Appropriate

WrittenModifications in

Method andProtocol

Step 2.28.1 1.00Days

No

No




Proceed to Step3: Formal Methods

Validation andReport Generation

StepStep 2.36 0.00

Days

Team IdentifiesModifications Needed to

Protocol and/or DraftMethod Which Require

AdditionalExperimentation

Step 2.29 0.50 Days

Team MembersConduct Additional

NecessaryExperiments

Step 2.32 5.00Days

Team LeaderCreates Summary

Report ofExperiments

Step 2.33 2.00Days

Team LeaderAssigns AdditionalLaboratory Workto TeamMembersStep 2.31 0.25

Days

AbbreviatedProtocol/Work

Plan GeneratedbyTeam LeaderStep 2.30 0.50

Days

Team Meets andReviews Additional

ExperimentalResults

Step 2.34 0.50Days

Is the Method Now“Validatable” via Method

and Protocol Modification?Step 2.35 0.25 Days

Yes

No

Total Estimated Time For Step 2 =75.0 Days3.8 Working Months



APPROACH 35

TABLE 4.1 Workflow Diagram for Step 2:Final Method Development and Trial Methods Validation

Estimated Step Description Duration Explanation

2.1 Validation team discusses 1.0 daydevelopment review report/summary as a group in open forum

2.2 Is method ready for 0.25 day Some criteria for a no-go decision may trial methods validation? include:

• Development work indicates that themethod does not satisfy significantcomponents of end-user requirements

• Insufficient development data tosupport the decisions to validate themethod

• A significantly better method wasdiscovered during step 1 based onliterature search, vendor contacts,previous experience of teammembers, etc.

If no, then

2.2.1 Validation team identifies 0.25 dayweaknesses in existing method

2.2.2 Validation team outlines 0.5 day[additional] development plan

2.2.3 Team leader creates 1.0 daydevelopment plan

2.2.4 Development plan circulated to 2.0 day See Appendix V for a template of an team for agreement example standard test method.

2.2.5 Development plan issued 20.0 dayand executed

2.2.6 Team leader compiles results 2.0 dayin a review report/summary

2.3 Team leader 1.0 daycreates/completes draft method

2.4 Team leader circulates draft 2.0 daymethod to appropriate team members for review and input

(Continued)



2.5 Team leader creates draft trial 2.0 days The trial methods validation protocolmethods validation protocol should:

• Match the appropriate USP methodcategory for use of method

• Mirror format and general content offinal methods validation protocol

• Have acceptance ranges versusacceptance criteria

• ID method performancecharacteristics which are appropriateto the validation

The trial or practice validation isdesigned to mimic most of thecomponents of the final validation. Itspurpose is to ensure the final validationis executed without deviation, if at allpossible. The trial validation protocoland acceptance criteria may be tweakedor modified to create the formalmethods validation protocol.

Note: See Appendix VI for a templateof an example methods validationprotocol.

2.6 Team members concur 1.0 dayon draft method content

2.7 Team members concur on 1.0 day draft trial methods validationprotocol

2.8 Team leader creates work 1.0 daybreakdown structure

2.9 Team leader creates project 1.0 day Project plan should be a simple plan document designed to give the trial

methods validation process some structure. It may include:

• Steps to be executed• Personnel assignments• Start and finish dates• Required output for each step• Current status and notes




APPROACH 37

Note: It should match/align with thevalidation SOP and/or protocol.

2.10 Team leader promulgates 1.0 dayproject plan to team members

2.11 Changes to project plan? 0.0 day If yes, team leader makes changes, andpromulgates to team members forreview.

2.12 Team members approve 0.5 dayproject plan

2.13 Team leader solicits feedback 1.0 dayfrom QA on draft trial methods validation protocol

2.14 Draft method finalized 1.0 dayas working method

2.15 Draft method included in trial 0.25 dayvalidation protocol

2.16 Trial methods validation 1.0 day If practical, the trial validation protocol is finalized protocol should give as much detail as

possible. This will ensure proper execution of the experiments. Some examples include:

• Detailed standard preparationinstructions

• Detailed sample preparationinstructions

• Detailed placebo preparationinstructions

• Detailed spiked placebo preparationinstructions

• Other clarifying instructions asappropriate

2.17 Team leader performs review 0.25 dayof trial methods validation protocol and validation SOP to ensure alignment

2.18 Trial methods validation 0.25 dayprotocol is issued and executed



(Continued)


2.19 Trial methods validation 0.25 dayproject plan is executed

2.20 Trial methods validation data 15.0 daycollected and reviewed as appropriate

2.21 Team leader compiles trial 2.0 day The purpose of this report is to methods validation data organize the trial validation data into in draft report format a final document whose format mimics

the desired final formal methodsvalidation report. This is done so thatduring step 3: formal methods validationand report generation, the final reportwill require minimum effort tocomplete.

The sections of the report should mirrorthe trial validation protocol and/orvalidation SOP.

Note: Remember, trial methodsvalidation ensures that no surprisesoccur during formal methods validation.Formal methods validation should bethe least complex and time-consumingportion of the validation.

2.22 Compare results obtained to 0.5 day In addition to being part of the body trial validation protocol of the report. A table should be acceptance criteria ranges generated which shows the desired

acceptance criteria, the desired ranges,and the experimentally derived results.

2.23 Team leader promulgates draft 2.0 daytrial methods validation report to team members for review and comment

2.24 Team leader solicits feedback 2.0 dayfrom QA on trial methods validation report format and content

2.25 Team leader schedules report 0.5 dayreview meeting with team members and QA





2.26 Trial validation report review 0.5 day In the meeting the following items meeting held with validation should be addressed:team members and QA • Were acceptance criteria within the

desired ranges?• Did the draft method perform as

expected?• Was the protocol lucid and did it

capture all instructions needed toexecute the trial validation?

2.27 Is the method validatable via 0.25 day If yes, proceed to step 3: Formal existing method and protocol? methods validation; otherwise continue

to step 2.28

2.28 Will modifications require 0.25 day If no, team leader makes appropriate further experiments? written modifications in method and

protocol and proceeds to step 3: formalmethods validation; otherwise proceedto step 2.29

2.29 Team identifies modifications 0.5 day An example of this includes forced needed to protocol and/or draft degradation studies. These are often method which require difficult to control and frequently additional experimentation require additional experimentation to

achieve the ~10% to ~30% degradationnecessary.

2.30 Abbreviated protocol/work 0.5 dayplan generated by team leader

2.31 Team leader assigns 0.25 dayadditional laboratory work to team members

2.32 Team members conduct 5.0 dayadditional necessary experiments

2.33 Team leader creates summary 2.0 dayreport of experiments

2.34 Team meets and reviews 0.5 dayadditional experimental results

APPROACH 39

(Continued)




2.35 Is the method now 0.25 day If no, return to step 2.29; otherwise validatable via method and continue to step 2.36protocol modification?

2.36 Proceed to step 3: formal 0.0 daymethods validation and report generation

Total estimated time for step 2 = 75.0days or 3.8 working months





5.1 BACKGROUND

At the conclusion of the tasks delineated in step 2: final method developmentand trial methods validation, any ambiguities associated with the method, themethods validation protocol, and the resources and approach taken to per-form the validation should no longer exist. The method is now ready to bevalidated via execution of the methods validation protocol and associatedproject plan.

It is now possible to meet the FDA’s expectation that methods validation isthe process of demonstrating that analytical procedure is suitable for itsintended use. The methods validation process will now truly be the plannedand systematic collection by the applicant of the validation data to supportanalytical procedures. Validation is also now truly the end game where fewsurprises and deviations will be encountered. The validation is executed witha formal, approved, and signed methods validation protocol in place whichhas been reviewed by the quality assurance unit.

At the conclusion of the experimental portion of formal methods validation,a methods validation report, which is supported by the experimental data, is

41


CHAPTER 5

STEP 3: FORMAL METHODSVALIDATION AND REPORTGENERATION


created, circulated, reviewed, and approved by all appropriate personnel. Thispackage is then forwarded to quality assurance for review, approval, archiving,and distribution. Any deviations from the protocol, including failures tomeeting acceptance criteria, must be fully investigated and appropriatelydocumented as would any out-of-specification (OOS) result.

Any decision to disregard data, initiate a retest or resample, or modifyacceptance criteria must be based on sound scientific data and scientificreasoning. Remember, deviations from the protocol no longer can be arm-waved away. The FDA expects you to scientifically address your failuresas you would any other laboratory investigation.

5.2 APPROACH

The basic elements of step 3: formal methods validation and report gene-ration are:

● Draft method is modified as necessary● Final methods validation protocol is created, reviewed, and approved by

all appropriate personnel including quality assurance ● Final project plan is created, reviewed, and approved by all appropriate

personnel● Formal methods validation is executed via implementation of methods

validation protocol and project plan● Data are collected, recorded, and reviewed as appropriate● Draft methods validation report is created● Draft methods validation report is reviewed by appropriate personnel● Draft report and supporting data package are forwarded to quality assur-

ance

Steps in this process are shown in Figure 5.1 and described in Table 5.1.

42 STEP 3: FORMAL METHODS VALIDATION AND REPORT GENERATION


APPROACH 43

Team LeaderMakes Final

Modifications toDraft Method

Step 3.1 1.00Days

Team LeaderMakes Final

Modifications toTrial Methods


Days

Team LeaderPromulgates FinalDraft Method andModified MethodsValidation Protocolfor Team Review

Step 3.3 2.00Days

Team Meets andAgrees on Final

Method andProtocol Format

and ContentStep 3.4 0.50

Days

Team LeaderCreates Final

MethodsValidation Protocol

with EmbeddedMethod

Step 3.5 0.25Days

Team LeaderForwards Protocolto QA for Approval

Step 3.6 2.00Days

Is QA satisfiedwith Protocol?Step 3.7 0.00

Days

Protocol Modifiedas Needed

Step 3.7.1 0.50Days

Team LeaderCreates Project

PlanStep 3.9. 0.50

Days

Signed ProtocolIssued forExecution

Step 3.8 0.25 Days

No

Yes

Team LeaderViews Protocol

and Project Planwith TeamMembers

Step 3.10 0.25Days

Team InventoriesReagents, Standards,Samples, and OtherSupplies to InsureAvailability DuringFormal Validation

Step 3.11 0.50 Days

Components of Step 3: Final Methods Validation and Report Generation

~1 to 2 Months

FIGURE 5.1 Workflow diagram for step 3: final methods validation and report generation.



MethodsValidation

Formally ExecutedVia Protocol andAdministered by

Project PlanStep 3.12 0.25

Days

Formal MethodsValidation Data

Collected,Recorded andReviewed asAppropriate

Step 3.13 15.00Days

Team LeaderCompiles Formal

MethodsValidation Data in

Draft ReportFormat

Step 3.14 5.00Days

Compare ResultsObtained to

Formal ValidationProtocol

AcceptanceCriteria RangesStep 3.15 0.50

Days

Team LeaderPromulgates DraftFormal MethodsValidation Reportto Team Members

for Review andComment

Step 3.16 2.00Days

Formal ValidationReport Review

Meeting Held withValidation TeamStep 3.18 0.50

Days

Team LeaderSchedules ReportReview Meeting

with TeamMembers 3.17

0.25 Days

Is Report ReadyFor Formal QA

Review?Step 3.19 0.25

Days

Identify Report as“Final Draft”

Step 3.20 0.25Days

Assemble orProvide Access to

All Notebooks,Chromatogramsand Other Raw

DataStep 3.21 0.25

Days

Forward FinalDraft Report andSupporting RawData to QA for

ReviewStep 3.22 0.25

Days

Team Leader andTeam Members

Makes AllAppropriate

ChangesStep 3.19.1 1.00

Days

No

Yse

Proceed to Step 4: Formal DataReview and

Report Issuance

Total Estimated Time For Step 3 =35.25Days

1.8 Working Months



APPROACH 45

TABLE 5.1 Workflow Diagram for Steps to Step 3:Final Methods Validation and Report Generation


3.1 Team leader makes final 1.0 daymodifications to draft method

3.2 Team leader makes final 2.0 daysmodifications to trial methodsvalidation protocol

3.3 Team leader promulgates 2.0 daysfinal draft method andmodified methods validationprotocol for team review

3.4 Team meets and agrees on 0.5 day Acceptance criteria must be set at thisfinal method and protocol point. Criteria should be fully attainable format and content based on results and experience gained

from trial validation work.

3.5 Team leader creates final 0.25 daymethods validation protocolwith embedded method

3.6 Team leader forwards 2.0 daysprotocol to QA for approval

3.7 Is QA satisfied with protocol? 0.0 day Protocol is modified as needed, and goback to step 3.6.

3.8 Signed protocol issued for 0.25 dayexecution

3.9 Team leader creates project 0.5 day Project plan should be a simple plan document designed to give the trial

methods validation process somestructure. It may include:

● Steps to be executed● Personnel assignments● Start and finish dates● Required output for each step● Current status and notes

Note: It should match/align with thevalidation SOP and/or protocol.

3.10 Team leader views protocol 0.25 dayand project plan with teammembers

(Continued)



3.11 Team inventories reagents, 0.5 daystandards, samples, and othersupplies to ensure availabilityduring formal validation

3.12 Methods validation formally 0.25 dayexecuted via protocol andadministered by project plan

3.13 Formal methods validation 15.0 days All deviations from protocol must be data collected, recorded, and approved and documented appropriately.reviewed as appropriate Deviations should be minimal at this

stage.

3.14 Team leader compiles formal 5.0 days The trial methods validation report shouldmethods validation data in draft be used as a template. Data are enteredreport format into the report as they are collected and

reviewed.

3.15 Compare results obtained to 0.5 day In addition to being part of the body offormal validation protocol the report, a table should be generatedacceptance criteria ranges showing the desired acceptance criteria

were met. As stated here, all deviationsfrom protocol must be approved anddocumented appropriately. Deviationsshould be minimal at this stage.

3.16 Team leader promulgates draft 2.0 daysformal methods validationreport to team members forreview and comment

3.17 Team leader schedules report 0.25 dayreview meeting with team

3.18 Formal validation report 0.5 day This should be a critical groupreview meeting held with discussion. End result should be a reportvalidation team that will stand on its own and be

understood by a reviewer who isunfamiliar with the development work.All work presented in the report musthave notebook or similar references toraw data. All data tables presented in thereport should be reproducible by manualcalculation check.

3.19 Is report ready for formal 0.25 dayQA review?




3.20 Identify report as final draft 0.25 day

3.21 Assemble or provide access 0.25 dayto all notebooks,chromatograms,and other raw data

3.22 Forward final draft report 0.25 dayand supporting raw data toQA for review

Proceed to step 4: formal datareview and report issuance

Total estimated time for step 3 � 35.25days or 1.8 working months.

APPROACH 47




48

6.1 BACKGROUND

The process of methods validation is complete when you have:

(1) Demonstrated that you have met all the acceptance criteria (2) Have clearly documented the results in a CGMP-compliant fashion(3) Have shown how you met the acceptance criteria in a final methods

validation report, including references to raw data, all of which hasbeen reviewed and approved by the appropriate personnel includingpeers, management, and QA.

Step 4: formal data review and report issuance, is the culmination of themethods validation process. It is a critical component to the process in that:

● It ensures that the reported results are supported by valid scientific data● The supporting data themselves are corrected as verified by peer and

quality assurance personnel review● It ensures the report is organized in a coherent fashion and can be under-

stood by any reviewer who is reasonably trained in the field of methodsvalidation


CHAPTER 6

STEP 4: FORMAL DATA REVIEW ANDREPORT ISSUANCE


● There are links to the results reported in the methods validation report sothat an outside reviewer can trace the reported results back to raw data

● It ensures the report has received the proper level of review and pos-sesses the corresponding management and QA approval signatures

● The report has been officially issued and controlled copies circulated tothe appropriate personnel

● Original copies of the report and all written and electronic data havebeen properly archived for easy retrieval in the future

Despite its criticality, step 4 is often not considered as a formal part of meth-ods validation by many organizations. Because of this, sufficient resourcesand emphasis are often not placed on this final component of the methodsvalidation process. This leads to difficulties during review by outside organi-zations and if difficulties are ever encountered by the end users followingtechnology transfer. Although tedious, proper data review and reportissuances procedures are the glue that binds the entire validation processtogether.

6.2 APPROACH

The basic elements for step 4 are:

● The quality assurance unit (QA) receives all supporting data and thedraft methods validation report

● QA performs formal report and data review; corrections made asnecessary

● Final methods validation report is generated with appropriate signaturecover sheet

● Report is circulated for approval by validation team members, QA, andmanagement

● Controlled copies of final signed methods validation report are made ● QA distributes controlled copies of report as appropriate● Methods validation report, notebooks, and all raw data are archived as

appropriate

Steps in this process are shown in Figure 6.1 and described in Table 6.1.Some details for each step are summarized in Table 6.1.

APPROACH 49


50 STEP 4: FORMAL DATA REVIEW AND REPORT ISSUANCE

QA Receives AllNotebooks,

Chromatograms andOther Raw Data Linkedto and Referenced inMethods Validation

ReportStep 4.1 0.50 Days

QA Receives FinalDraft Report

Step 4.2 0.25Days

QA Obtains FinalCopy of Methods


Days

QA ObtainsCurrent MethodsValidation SOP

RevisionStep 4.4 0.25

Days

QA ReviewsMethods

Validation Reportand Flags Any

ApparentDiscrepanciesStep 4.5 5.00

Days

Were DiscrepanciesIdentified?

Step 4.6 0.00 Days

No

FlagDiscrepancies andReview with Team

Lead andAppropriate Team

MembersStep 4.6.1 0.25

Days

Corrections andModification to theReport Discussedwith QA and Made

as AppropriateStep 4.6.2 2.00

Days

Formal Data Review and Report Issuance~1 to 2 Months

Yes

FIGURE 6.1 Workflow diagram of step 4: formal data review and report issuance.


APPROACH 51

Cover andSignature PagesAre GeneratedStep 4.10 0.25

Days

Final Electronicand Hard Copies

of Data andReports are

Placed in QA'sCustody

Step 4.7 0.50Days

Reports and DataEntered into

Formal DocumentControl SystemStep 4.9 1.00

Days

QA Takes FormalControl of Reports

and Raw DataStep 4.8 0.25

Days

Report isCirculated for AppropriateManagerial

ReviewStep 4.11 0.25

Days

Managers ReviewMethods

Validation ReportFor Any Apparent

DiscrepanciesStep 4.12 10.00

Days

Were DiscrepanciesIdentified?

step 4.13 0.00 Days

FlagDiscrepancies andReview with Team

Lead andAppropriate Team

MembersStep 4.13.1 2.00

Days

Corrections andModifications to

the ReportDiscussed with

QA/Managementand Made asAppropriate

Step 4.13.2 2.00Days

Yes

No

Manager's Sign Signature Page

Approving ReportStep 4.14 2.00

Documents

VALIDATING CHROMATOGRAPHIC METHODS - idqc-hcm.gov.vn...VALIDATING CHROMATOGRAPHIC METHODS A Practical Guide DAVID M. BLIESNER JWUS_VC-Blies_FM.qxd 7/17/2006 4:02 PM Page iii