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

Table of values (x,y)

Linearity
Calculated analyte in mg/mL

0.040

0.035 of an analyte in a material x y

0.030 Reference Calculated
# material mg/ml mg/ml
0.025
1 0.0100 0.0101
0.020
2 0.0150 0.0145
0.015
Reference material mg/ml 3 0.0200 0.0210
0.010
4 0.0250 0.0260
0.01 0.015 0.02 0.025 0.03 0.035 0.04
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

Limit of Detection Peak B

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