PHARMACEUTICS

1. ICH GUIDELINES FOR
METHOD VALIDATION

2. BY
SUMEL ASHIQUE
M.PHARM ( PHARMACEUTICS )
ISF COLLEGE OF PHARMACY
.MOGA (PB)

3. CONTENTS
 METHOD VALIDATION
 PURPOSE OF METHOD VALIDATION
 ICH AND USP GUIDELINES

4.  METHOD VALIDATION :
Method validation is the process of demonstrating that analytical
procedures are suitable for their intended use and that they support the
identity, strength, quality, purity and potency of the substances
in products.
 Method validation is primarily concerned with:
Identification of the sources of potential errors
Quantification of the potential errors in the method
An method validation describes in mathematical and quantifiable t e r
ms t h e p e rf o rma n c e c h a ra c t e ri s t i cs of an a s sa y
4

5. Examples of Methods That Require
Validation Documentation
 Chromatographic Methods - HPLC, GC, TLC, GC/MS, etc.
Pharmaceutical Analysis - In support of CMC.
Bioanalytical Analysis - In support of PK/PD/Clinical Studies.
 Spectrophotometric Methods – UV/VIS, IR, AAS, XRD, ICP-MS,
AAS, XRF, etc
 Particle Size Analysis Methods - Laser, Microscopic, Sieving, SEC, etc.
 Automated Analytical Methods - Robots, AutomatedAnalysis.

6. Considerations Prior to
Method Validation
 Suitability of Instrument
 Status of Qualification and Calibration
 Suitability of Materials
 Status of Reference Standards, Reagents, Placebo
Lots
 Suitability of Analyst
 Status of Training and Qualification Records
 Suitability of Documentation
 Written analytical procedure and proper approved
protocol
 with pre-established acceptance criteria

7. Purpose of Method
Validation
 Identification of Sources and Quantitation of Potential errors
 Determination if Method is Acceptable for IntendedUse
 Establish Proof that a Method Can be Used for DecisionMaking
 Satisfy Requirements

8. Published Validation Guidelines
 1978 Current Good Manufacturing Practices (cGMPs)
 1987 FDA Validation Guideline
 1989 Supplement 9 to USP XXI
 1994 CDER Reviewer Guidance: Validation
of Chromatographic Method
 1995 ICH Validation Definitions:
Q2A, Text on Validation of Analytical procedures
 1997 ICH Validation Methodology:
Q2B, Validation of Analytical Procedures: Methodology
 1999 Supplement 10 to USP 23 <1225>: Validation of Compendial Methods
 1999 CDER “Bioanalytical Method Validation for Human Studies”
 2000 CDER Draft “Analytical Procedures and Method Validation”

9. Today’s Validation
Requirements
ICH/USP
GMPs
(legal) FDA

10. ICH/USP Validation
Requirements &
Parameters
ICH
 Specificity
 Linearity
 Range
 Accuracy
 Precision
 Repeatability
 Intermediate Precision
 Reproducibility
 Limit of Detection
 Limit of Quantitation
USP
 Specificity
 Linearity and Range
 Accuracy
 Precision
 Limit of Detection
 Limit of Quantitation
 Ruggedness
 Robustness

11. Specificity/Selectivity
 Ability of an analytical method to measure the analyte freefrom
interference due to other components.
Specificity is the ability to assess unequivocally the analyte in the presence of components
which may be expected to be present. Typically these might include impurities, degradants,
matrix, etc.
Purity Tests: to ensure that all the analytical procedures performed allow an accurate
statement of the content of impurities of an analyte, i.e. related substances test, heavy
metals, residual solvents content, etc.
Assay (content or potency): to provide an exact result which allows an accurate statement
on the content or potency of the analyte in a sample.
 Selectivity describes the ability of an analytical method todifferentiate
various substances in a sample

12. Linearity
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 Ability of an assay to elicit a direct and
proportional response to changes in
analyte concentration.

13. Linearity Should be Evaluated
 By Visual Inspection of plot of signals vs. analyte
concentration
 By Appropriate statistical methods
 Linear Regression (y = mx + b)
 Correlation Coefficient, y-intercept (b), slope(m)
 Acceptance criteria: Linear regression r2 >0.95
Requires a minimum of 5 concentration levels

14. Range
 The specified range is normally derived from linearity studies and depends on the intended application of
the procedure. It is established by confirming that the analytical procedure provides an acceptable degree of
linearity, accuracy and precision when applied to samples containing amounts of analyte within or at the
extremes of the specified range of the analytical procedure.
 Acceptable range having linearity, accuracy, precision.
 For Drug Substance & Drug product Assay
 80 to 120% of test Concentration
 For Content Uniformity Assay
 70 to 130% of test Concentration
 For Dissolution Test Method
 +/- 20% over entire Specification Range
 For Impurity
 From MDL to 100% of Impurity Specification Limit

15. Accuracy
 Closeness of the test results obtained by the method to the true value.
 Should be established across specified range of analytical procedure.
 Should be assessed using a minimum of 3 concentration levels, each in triplicate
(total of 9 determinations)
 Should be reported as:
 Percent recovery of known amount added or
 The difference between the mean assay result and the accepted value.

16. Precision
 The closeness of agreement (degree of scatter) between a series of
measurements obtained from multiple samplings of the same homogeneous
sample.
 Should be investigated using homogeneous, authentic samples.
Precision Considered at 3 Levels :
 Repeatability
 Intermediate Precision
 Reproducibility

17. Repeatability
 Express the precision
under the same
operating conditions
over a short interval of
time.
 Also referred to as
Intra-assay precision
Should be assessed
using minimum of 9
determinations
(3 concentrations/ 3
replicates) or
Minimum of 6
determinations at the
100% level.

18. Reproducibility
 Definition: Ability reproduce data
within the predefined precision
 Determination: SD, RSD and confidence interval
 Repeatability test at two different labs.

19. Detection Limit (LOD)/
Quantitation Limit (LOQ)
 LOD
Lowest amount of analyte in a
sample that can be detected
but not necessarily
quantitated.
Estimated by Signal to Noise
Ratio of 3:1.
LOQ
Lowest amount of analyte
in a sample that canbe
quantified with suitable
accuracy and precision.
Estimated by Signal to
Noise Ratio of 10:1.

20.  Definition: Capacity to remain unaffected by small butdeliberate
variations in method parameters
 Determination: Comparison results under differingconditions
with precision under normal conditions
 Examples of typical variations in LC
 Influence of variations of pH in a mobile phase
 Influence of variations in mobile phase composition
 Different columns (different lots and/or suppliers)
 Temperature
 Flow rate
Robustness