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