Module 1,, Part 4: QC-related validation Slide 1 of 28 © WHO – EDM – 1/2002

Validation Part 4:

QC-related validation

Supplementary Training Modules on Good Manufacturing Practice

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Validation

Introduction

Why is analytical monitoring necessary?

What is the purpose of analytical validation?

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Validation

Objectives

To introduce the concepts of : Protocol development

Instrument qualification

Analytical procedure

Extent of validation

Method transfer

Chemical and physical, biological, and microbiological test validation

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Validation of analytical procedures requires: Qualified and calibrated instruments

Documented methods

Reliable reference standards

Qualified analysts

Sample integrity

Validation

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Validation protocol for analytical method Statement of purpose and scope Responsibilities Documented test method List of materials and equipment Procedure for the experiments for each parameter Statistical analysis Acceptance criteria for each performance parameter

Validation

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Qualification of the instrument

Make, model and maker’s manual

Modifications

Installation and operational qualification

Calibration programs

Maintenance schedules

Validation

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Characteristics of analytical procedures (1) Accuracy

Precision

Repeatability

Reproducibility

Validation

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Inaccurate &imprecise

Inaccurate butprecise

Accurate butimprecise

Validation

Relationship between accuracy and precision

Accurate AND Precise

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Characteristics of analytical procedures (2)

Ruggedness Robustness Variability caused by:

Day-to-day variations Analyst-to-analyst Laboratory-to-laboratory Instrument-to-instrument Chromatographic column-to-column Reagent kit-to-kit Instability of analytical reagents

Validation

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Characteristics of analytical procedures (3)

Linearity and range

Specificity

Sensitivity

Limit of detection

Limit of quantitation

Validation

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Linearity of an analyte in a material

0.010

0.015

0.020

0.025

0.030

0.035

0.040

0.01 0.015 0.02 0.025 0.03 0.035 0.04

Reference material mg/mlCalculated analyte in mg/mL

Table of values (x,y)

x y # Reference

material mg/mlCalculated

mg/ml

1 0.0100 0.0101

2 0.0150 0.0145

3 0.0200 0.0210

4 0.0250 0.0260

5 0.0300 0.0294

6 0.0400 0.0410

Validation

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Linearity Statistics Intercept -0.0002

Limit of Linearity and Range 0.005 – 0.040 mg/mL

Slope 1.0237

Correlation coefficient Pearson 0.9978 Olkin and Pratt 0.9985

Relative procedure standard deviation 3.4%

Validation

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LOQ, LOD and SNR

Limit of Quantitation

Limit of Detection

Signal to Noise Ratio

noise

Peak ALOD

Peak BLOQ

Baseline

Validation

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Different classes of analytical tests

Class A: To establish identity

Class B: To detect and quantitate impurities

Class C: To determine quantitatively the concentration

Class D: To assess the characteristics

Validation

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* A degree of bias may be allowed

Characteristic A B quant.

B Limit test

C D

Accuracy 

  X   X X*

Precision   X   X X

Robustness X X X X X

Linearity and range

  X   X X

Specificity X X X X X

Limit of detection   X    

Limit of quantitation

  X      

Validation

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Extent of validation

New methods require complete validation

Pharmacopoeial methods require partial validation (or verification)

Significant changes mean partial revalidation equipment changes formula changed changed suppliers of critical reagents

Validation

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Analytical method transfer Method transfer protocol and procedure

precision accuracy ruggedness

Written and approved specific test method

Proficiency check

Formal acceptance by new laboratory

Validation

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Chemical laboratory validation requirements (1)

Balances Chromatography

HPLC, HPTLC, GC, TLC Dissolution or disintegration apparatus Karl Fischer moisture determination Melting, softening or freezing point apparatus Ovens, refrigerators, incubators

Validation

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Chemical laboratory validation requirements (2) pH meter

Polarimeter - optical rotation

Refractometer

Spectrophotometer UV/Vis, IR, FTIR, Raman, AA

Timers

Viscometer

Volumetric equipment

Validation

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Validation

Typical validation of HPCL assay (1)

System suitability (performance check) system precision column efficiency symmetry factor capacity factor

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Validation

Typical validation of HPLC assay (2) Method validation

specificity accuracy precision linearity robustness

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Biological assays Can be difficult to "validate"

"Validity" on a case by case basis

Strictly adhere to the Biological Testing monographs in pharmacopoeias

Validation

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Microbiological testing requiring validation

Microbial limit testing

Microbial count

Sterility testing

Preservative effectiveness testing

Environmental monitoring program

Biological testing

Validation

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Validation of microbial test procedures (1)

Virtually impossible to completely validate test procedures for every microorganism

Neutralize /inactivate inhibitory substances, or dilute

Periodic media challenge

Media QC

Reliable methods

Validation

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Validation of microbial test procedures (2)

Incubation temperature and time

Media may not grow all microorganisms

Variations in media may affect recovery

Inhibitory disinfectants or preservatives

Sample

procedures

handling, storage, transport

Validation

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Microbiological viable count method validation (1) Methods

pour plate / spread plate membrane filtration Most Probable Number

Sample size

Test dilution

Inoculation size

Validation

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Microbiological viable count method validation (2)

Membrane filtration conditions

Incubation conditions

Acceptance criteria

Validation

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Sterility testing validation

requirements

Media growth promotion, sterility, pH

Product validation

Stasis testing

Environmental monitoring

Negative controls

Challenge organisms

Validation