Analytical method validation is a process of documenting/ proving that an analytical method provides analytical data acceptable for the intended use.After the development of an analytical procedure, it is must important to assure that the procedure will consistently produce the intended a precise result with high degree of accuracy. The method should give a specific result that may not be affected by external matters. This creates a requirement to validate the analytical procedures. The validation procedures consists of some characteristics parameters that makes the method acceptable with addition of statistical tools.

1. Analytical method validation as per ICH and USP
SHREYAS B.R
1ST MPHARM QA

2. CONTENTS
 Introduction
 Objective
 Types of analytical procedures to be validated
 Validation parameters as per ICH and USP
 Conclusion
 Reference

3. INTRODUCTION
 Validation is the documented act of proving that any procedure,
process, equipment, material, activity or system actually leads to the
expected result.
 Analytical method validation is a process of documenting/proving
that an analytical method provides analytical data acceptable for the
intended use.
 A pharmaceutical drug product must meet all its specifications
through out its shelf-life.
 The method of analysis used must be validated. This is required to
ensure the product’s safety and efficacy through out all phases of its
shelf-life.

4. Objective
 The main objective of analytical validation is to ensure that a selected
analytical procedure will give reproducible and reliable results that are
adequate for the intended purpose.
 This is applicable to all the procedure either pharmacopoeial or non
pharmacopoeial.

5. Types of analytical procedures to be validated
 The required validation parameters also termed “ analytical
performance characteristics”, depends upon the type of analytical
method.
 Pharmaceutical analytical methods are characterized into 5 general
types
• Identification tests
• Potency assays
• Limit tests for the control of impurities
• Impurity tests- quantitative
• Specific tests

6. Validation parameters as per ICH/USP
 USP
• Specificity
• Linearity or range
• Accuracy
• Precision
• Limit of detection
• Limit of quantitation
• Ruggedness
• Robustness
 ICH
• Specificity
• Linearity
• Range
• Accuracy
• Precision
• Limit of detection
• Limit of quantitation
• Robustness

7. 1. Accuracy
Definition: “The accuracy of an analytical procedure is the
closeness of agreement between the values that are accepted either
as conventional true values or an accepted reference value and the
value found”.
Determination
 Assay
i. Drug substance
ii. Drug product
 Impurities (quantitation)

8.  Recommended data: Assessed by 9 determinations over a
minimum of 3 concentration levels covering a specified range.
 Limit
i. Typical accuracy of the recovery of the drug substance is
expected to be about 99 – 101%.
ii. Typical accuracy of the recovery of the drug product is
expected to be about 98 – 102%.

9. 2. Precision
Definition: “The closeness of agreement (degree of scatter)
between a series of measurements obtained from multiple
samplings of the same homogeneous sample”.
Precision includes:
 Repeatability
 Intermediate Precision
 Reproducibility

10.  Repeatability
• Repeatability expresses the precision under the same operating
conditions over a short interval of time.
• Repeatability should be assessed using a minimum of 9
determinations covering the specified range.
 Intermediate precision
• Intermediate precision expresses variations within laboratories,
such as different days, different analysts, different equipment etc.
 Reproducibility
• Reproducibility expresses the precision between laboratories.

11.  Following parameters should be reported
a. Standard deviation.
b. Relative standard deviation.
Concentration
µg/ml
Absorbance SD and %RSD
8 0.337 0.00041,
1.223%0.348
0.341
12 0.575 0.0106,
1.815%0.583
0.596
20 0.967 0.0091,
0.933%0.985
0.978
0
0.2
0.4
0.6
0.8
1
1.2
4 8 12 16 20 24
8
8
8
12
12
12
20
20
20

12. 3. Limit of detection
It is the lowest amount of analyte in a sample which can be detected but
not necessarily quantitated.
4. Limit of quantitation
It is the lowest amount of analyte in a sample which can be
quantitatively determined with suitable precision and accuracy.

13. Determination of LOD and LOQ
 Limit of detection
o Method
 Based on visual examination.
 Based on standard deviation
of response and slope.
 Signal to noise ratio 2:1 or 3:1
 Limit of quantitation
o Method
 Based on visual examination.
 Based on standard deviation
of response and slope.
 Signal to noise ratio 10:1

14. 5. Specificity
Definition: Specificity is the ability to assess unequivocally the analyte
in presence of components which may be expected to be present.
Determination
• Identification tests
• Assay and impurity test(s)
Impurities are available
Impurities are not available

15. 6. Linearity
Definition: The Ability of the method to obtain test results that are
directly proportional to concentration within a given range.
Method: dilution of stock solution/separate weightings
 Minimum 5 concentrations are used.

16. 7. Range
Definition: The interval between the upper and lower concentrations
of analyte in the sample that have been demonstrate to have a
suitable level of precision, accuracy, and linearity.
 Established by confirming that the method provides acceptable
degree of linearity, accuracy, and precision.
 Specific range dependent upon intended application of the
procedure.

17.  Assay: 80 to 120% of test concentration.
 Content uniformity: 70 to 130% of test concentration.
 Dissolution: 20% to 120%
 Impurities reporting level: 120% of specification limit (with
respect to test concentration of API)

18. 8. Ruggedness
Definition: The ruggedness of an analytical method is the degree of
reproducibility of test results obtained by the analysis of the same
samples under a variety of conditions, such as different laboratories,
different analysts, different instruments, different days, etc.
 Certain may include-
i. Source
ii. Concentration and stability of solution
iii. Heating rate
iv. Column temperature
v. Humidity

19. 9. Robustness
Definition: “The robustness of an analytical procedure is a measure of
its capacity to remain unaffected by small, but deliberate variations in
method parameters and provides an indication of its reliability during
normal usage”.
Determination:
The evaluation of robustness should be considered during the
development phase and depends on the type of procedure under study.

20. • Variations may include:
 stability of analytical solution
 variation of pH in a mobile phase
 different column (lot/supplier)
 temperature
 flow rate

21. 10. System suitability
 System suitability testing is an integral part of many analytical
procedures.
 The tests are based on the concept that the equipment, electronics,
analytical operations and samples to be analyzed constitute an
integral system that can be evaluated as such.
 System suitability testing has been recommended by USP in
HPLC procedures.

22. Conclusion
 When the method is properly validated consistent, reliable and
accurate results are obtained.
 Analytical method validation is an important analytical tool to ensure
the accuracy and specificity of the analytical procedures with a
precise agreement.
 Validation of analytical methods is also required by regulations.
 Hence it is very important to validate any analytical method that has
been developed.

23. References
1. Gupta P. C method validation of analytical procedures. pharmatutor. 2015;3(1):32-39.
2. Lopez P, Buffoni E, Pereira F, Vilchez Quero J. Analytical Method Validation. Wide Spectra
of Quality Control. 2011;:1-19.
3. [Internet]. Who.int. 2016 [cited 2 March 2019].
https://www.who.int/medicines/areas/quality_safety/quality_assurance/Guideline_Validation
_AnalyticalMethodValidationQAS16-671.pdf
4. Daksh S. VALIDATION OF ANALYTICAL METHODS – STRATEGIES &
SINGFICANCE. International Journal of Research and Development in Pharmacy and Life
Sciences. 2015;4(3):1489-1497.
5. [Internet]. Ich.org. [cited 2 March 2019].
https://www.ich.org/fileadmin/PublicWebSite/ICH_Products/Guidelines/Quality/Q2
_R1/Step4/Q2_R1Guideline.pdf
6. Sharma P. validation in pharmaceutical industry. 2nd ed. Delhi: Vandana
publications; 2007. P: 425-448

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