Analytical Validation Method for laboratory

Analytical validation

1. Module 1,, Part 4: QC-related validation Slide 1of28 © WHO –EDM –1/2002
Validation Part 4:
QC-related validation
Supplementary Training Modules on
Good Manufacturing Practice

2. Module 1,, Part 4: QC-related validation Slide 2of28 © WHO –EDM –1/2002
Validation
Introduction
Why is analytical monitoring necessary?
What is the purpose of analytical validation?

3. Module 1,, Part 4: QC-related validation Slide 3of28 © 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

4. Module 1,, Part 4: QC-related validation Slide 4of28 © WHO –EDM –1/2002
Validation of analytical procedures
requires:
 Qualified and calibrated instruments
 Documentedmethods
 Reliablereferencestandards
 Qualifiedanalysts
 Sampleintegrity
Validation

5. Module 1,, Part 4: QC-related validation Slide 5of28 © WHO –EDM –1/2002
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

6. Module 1,, Part 4: QC-related validation Slide 6of28 © WHO –EDM –1/2002
Qualification of the instrument
Make, model and maker’s manual
Modifications
Installation and operational qualification
Calibration programs
Maintenance schedules
Validation

7. Module 1,, Part 4: QC-related validation Slide 7of28 © WHO –EDM –1/2002
Characteristics of analytical
procedures (1)
Accuracy
Precision
Repeatability
Reproducibility
Validation

8. Module 1,, Part 4: QC-related validation Slide 8of28 © WHO –EDM –1/2002Inaccurate &
imprecise Inaccurate but
precise Accurate but
imprecise
Validation
Relationship between accuracy and
precision
Accurate AND Precise

9. Module 1,, Part 4: QC-related validation Slide 9of28 © WHO –EDM –1/2002
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

10. Module 1,, Part 4: QC-related validation Slide 10of28 © WHO –EDM –1/2002
Characteristics of analytical procedures (3)
Linearityandrange
Specificity
Sensitivity
Limitofdetection
Limitofquantitation
Validation

11. Module 1,, Part 4: QC-related validation Slide 11of28 © WHO –EDM –1/2002Linearity
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/ml
Calculated analyte in mg/mL
Table of values (x,y)
x y
#
Reference
material mg/ml
Calculated
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

12. Module 1,, Part 4: QC-related validation Slide 12of28 © WHO –EDM –1/2002
Linearity Statistics
Intercept -0.0002
Limit of Linearity and Range0.005 –0.040 mg/mL
Slope 1.0237
Correlation coefficient
Pearson 0.9978
Olkin and Pratt 0.9985
Relative procedure standard
deviation 3.4%
Validation

13. Module 1,, Part 4: QC-related validation Slide 13of28 © WHO –EDM –1/2002
LOQ, LOD and SNR
Limit of Quantitation
Limit of Detection
Signal to Noise Ratio
noise
Peak A
LOD
Peak B
LOQ
Baseline
Validation

14. Module 1,, Part 4: QC-related validation Slide 14of28 © WHO –EDM –1/2002
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

15. Module 1,, Part 4: QC-related validation Slide 15of28 © WHO –EDM –1/2002
* A degree of bias may be allowed
CharacteristicA 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

16. Module 1,, Part 4: QC-related validation Slide 16of28 © WHO –EDM –1/2002
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

17. Module 1,, Part 4: QC-related validation Slide 17of28 © WHO –EDM –1/2002
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

18. Module 1,, Part 4: QC-related validation Slide 18of28 © WHO –EDM –1/2002
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

19. Module 1,, Part 4: QC-related validation Slide 19of28 © WHO –EDM –1/2002
Chemical laboratory validation requirements (2)
pH meter
Polarimeter -optical rotation
Refractometer
Spectrophotometer UV/Vis, IR, FTIR, Raman, AA
Timers
Viscometer
Volumetric equipment
Validation

20. Module 1,, Part 4: QC-related validation Slide 20of28 © WHO –EDM –1/2002
Validation
Typical validation of HPCL assay (1)
System suitability (performance check)
system precision
column efficiency
symmetry factor
capacity factor

21. Module 1,, Part 4: QC-related validation Slide 21of28 © WHO –EDM –1/2002
Validation
Typical validation of HPLC assay (2)
Method validation
specificity
accuracy
precision
linearity
robustness

22. Module 1,, Part 4: QC-related validation Slide 22of28 © WHO –EDM –1/2002
Biological assays
Can be difficult to "validate"
"Validity" on a case by case basis
Strictly adhere to the Biological Testing
monographs in pharmacopoeias
Validation

23. Module 1,, Part 4: QC-related validation Slide 23of28 © WHO –EDM –1/2002
Microbiologicaltesting requiring validation
Microbiallimittesting
Microbialcount
Sterilitytesting
Preservativeeffectivenesstesting
Environmentalmonitoringprogram
Biologicaltesting
Validation

24. Module 1,, Part 4: QC-related validation Slide 24of28 © WHO –EDM –1/2002
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

25. Module 1,, Part 4: QC-related validation Slide 25of28 © WHO –EDM –1/2002
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

26. Module 1,, Part 4: QC-related validation Slide 26of28 © WHO –EDM –1/2002
Microbiological viable count method
validation (1)
 Methods
pour plate / spread plate
membrane filtration
Most Probable Number
 Sample size
 Test dilution
 Inoculation size
Validation

27. Module 1,, Part 4: QC-related validation Slide 27of28 © WHO –EDM –1/2002
Microbiological viable count method
validation (2)
Membrane filtration conditions
Incubation conditions
Acceptance criteria
Validation

28. Module 1,, Part 4: QC-related validation Slide 28of28 © WHO –EDM –1/2002
Sterility testing validation requirements
Media growth promotion, sterility, pH
Product validation
Stasis testing
Environmental monitoring
Negative controls
Challenge organisms
Validation