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

ObjectivesTo 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 XRobustness X X X X XLinearity and range

  X   X XSpecificity X X X X XLimit 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