Supplementary Training Modules on

Good Manufacturing Practice

Validation Part 4:
QC-related validation

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

 Validation

Introduction
 Why is analytical monitoring necessary?
 What is the purpose of analytical validation?

Module 1,, Part 4: QC-related validation Slide 2 of 28 © WHO – EDM – 1/2002

 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
Module 1,, Part 4: QC-related validation Slide 3 of 28 © WHO – EDM – 1/2002
 Validation

Validation of analytical procedures

requires:
 Qualified and calibrated instruments
 Documented methods
 Reliable reference standards
 Qualified analysts
 Sample integrity

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

 Validation
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

Module 1,, Part 4: QC-related validation Slide 5 of 28 © WHO – EDM – 1/2002

 Validation

Qualification of the instrument

 Make, model and maker’s manual
 Modifications
 Installation and operational qualification
 Calibration programs
 Maintenance schedules

Module 1,, Part 4: QC-related validation Slide 6 of 28 © WHO – EDM – 1/2002

 Validation

Characteristics of analytical
procedures (1)
 Accuracy
 Precision
 Repeatability
 Reproducibility

Module 1,, Part 4: QC-related validation Slide 7 of 28 © WHO – EDM – 1/2002

 Validation
Relationship between accuracy and
precision

Inaccurate &
imprecise

Inaccurate but Accurate but Accurate AND Precise

precise imprecise
Module 1,, Part 4: QC-related validation Slide 8 of 28 © WHO – EDM – 1/2002
 Validation
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

Module 1,, Part 4: QC-related validation Slide 9 of 28 © WHO – EDM – 1/2002

 Validation
Characteristics of analytical procedures (3)
 Linearity and range
 Specificity
 Sensitivity
 Limit of detection
 Limit of quantitation

Module 1,, Part 4: QC-related validation Slide 10 of 28 © WHO – EDM – 1/2002

 Validation

Linearity Table of values

Calculated analyte in mg/mL

0.040

of an analyte in a material (x,y)

0.035

0.030 x y
 
Reference Calculated
0.025 # material mg/ml mg/ml
0.020 1 0.0100 0.0101
0.015 Reference material mg/ml 2 0.0150 0.0145
0.010 3 0.0200 0.0210
0.01 0.015 0.02 0.025 0.03 0.035 0.04
4 0.0250 0.0260
5 0.0300 0.0294
6 0.0400 0.0410
Module 1,, Part 4: QC-related validation Slide 11 of 28 © WHO – EDM – 1/2002
 Validation
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%
Module 1,, Part 4: QC-related validation Slide 12 of 28 © WHO – EDM – 1/2002
 Validation

LOQ, LOD and SNR

 Limit of Quantitation
Peak B
 Limit of Detection LOQ
 Signal to Noise Ratio
Peak A
LOD

Baseline noise

Module 1,, Part 4: QC-related validation Slide 13 of 28 © WHO – EDM – 1/2002

 Validation

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

Module 1,, Part 4: QC-related validation Slide 14 of 28 © WHO – EDM – 1/2002

 Validation
Characteristic A B B C D
quant. Limit
test
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      
* A degree of bias may be allowed
Module 1,, Part 4: QC-related validation Slide 15 of 28 © WHO – EDM – 1/2002
 Validation
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

Module 1,, Part 4: QC-related validation Slide 16 of 28 © WHO – EDM – 1/2002

 Validation

Analytical method transfer

 Method transfer protocol and procedure
 precision
 accuracy
 ruggedness
 Written and approved specific test method
 Proficiency check
 Formal acceptance by new laboratory

Module 1,, Part 4: QC-related validation Slide 17 of 28 © WHO – EDM – 1/2002

 Validation
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

Module 1,, Part 4: QC-related validation Slide 18 of 28 © WHO – EDM – 1/2002

 Validation
Chemical laboratory validation requirements (2)
 pH meter
 Polarimeter - optical rotation
 Refractometer
 Spectrophotometer UV/Vis, IR, FTIR, Raman, AA
 Timers
 Viscometer
 Volumetric equipment

Module 1,, Part 4: QC-related validation Slide 19 of 28 © WHO – EDM – 1/2002

 Validation
Typical validation of HPCL assay (1)
 System suitability (performance check)
 system precision
 column efficiency
 symmetry factor
 capacity factor

Module 1,, Part 4: QC-related validation Slide 20 of 28 © WHO – EDM – 1/2002

 Validation
Typical validation of HPLC assay (2)
 Method validation
 specificity
 accuracy
 precision
 linearity
 robustness

Module 1,, Part 4: QC-related validation Slide 21 of 28 © WHO – EDM – 1/2002

 Validation
Biological assays
 Can be difficult to "validate"
 "Validity" on a case by case basis
 Strictly adhere to the Biological Testing
monographs in pharmacopoeias

Module 1,, Part 4: QC-related validation Slide 22 of 28 © WHO – EDM – 1/2002

 Validation
Microbiological testing requiring validation
 Microbial limit testing
 Microbial count
 Sterility testing
 Preservative effectiveness testing
 Environmental monitoring program
 Biological testing

Module 1,, Part 4: QC-related validation Slide 23 of 28 © WHO – EDM – 1/2002

 Validation
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

Module 1,, Part 4: QC-related validation Slide 24 of 28 © WHO – EDM – 1/2002

 Validation
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

Module 1,, Part 4: QC-related validation Slide 25 of 28 © WHO – EDM – 1/2002

 Validation
Microbiological viable count method
validation (1)
 Methods
 pour plate / spread plate
 membrane filtration
 Most Probable Number
 Sample size
 Test dilution
 Inoculation size

Module 1,, Part 4: QC-related validation Slide 26 of 28 © WHO – EDM – 1/2002

 Validation
Microbiological viable count method
validation (2)
 Membrane filtration conditions
 Incubation conditions
 Acceptance criteria

Module 1,, Part 4: QC-related validation Slide 27 of 28 © WHO – EDM – 1/2002

 Validation

Sterility testing validation requirements

 Media growth promotion, sterility, pH
 Product validation
 Stasis testing
 Environmental monitoring
 Negative controls
 Challenge organisms

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