This document discusses analytical method validation as per ICH and USP guidelines. It defines validation as establishing documentary evidence that a procedure maintains compliance. Method validation involves demonstrating that an analytical procedure is suitable for its intended purpose by testing parameters such as accuracy, precision, specificity, detection limit, quantitation limit, linearity, range, ruggedness and robustness. It also discusses the different types of analytical procedures that require validation including identification tests, quantitative impurity tests, limit tests and assays.

1. Analytical Methods Validation
as per ICH & USP Guidelines
Mr. Ganesh B. Nigade,
Assistant Professor,
Dept. of Pharmaceutical Chemistry
PDEA’s S G R S College of Pharmacy, Saswad

2. Contents
• Introduction
• QSEM
• Validation
• Analytical Method validation
• Types of Analytical Procedures to be Validated
• Validation Parameters
• References

3. Introduction
International Council for Harmonisation (ICH):-
 The International Council for Harmonisation of Technical
Requirements for Pharmaceuticals for Human Use.
 (ICH) is unique in bringing together the regulatory authorities and
pharmaceutical industry to discuss scientific and technical aspects
of pharmaceuticals and develop ICH guidelines.
 Since its inception in 1990, ICH has gradually evolved, to respond to
increasingly global developments in the pharmaceutical sector and
these ICH guidelines are applied by a growing number of regulatory
authorities.
 ICH's mission is to achieve greater harmonisation worldwide to
ensure that safe, effective and high quality medicines are
developed, and registered and maintained in the most resource
efficient manner whilst meeting high standards.
• ICH Secretariat office is at Geneva, Switzerland

4. The ICH topics are divided into the four categories- QSEM
Quality Guidelines
in the Quality area including stability
studies, impurities testing and a
more flexible approach to
pharmaceutical quality based on
GMP risk management.
Safety Guidelines
safety Guidelines to uncover potential
risks like carcinogenicity, genotoxicity
and reprotoxicity
non-clinical testing strategy for
assessing the QT interval prolongation
liability.
Efficacy Guidelines
the Efficacy heading is concerned
with the design, conduct, safety and
reporting of clinical trials.
It also covers novel types of
medicines derived from
biotechnological processes and the
use of pharmacogenetics/genomics
techniques to produce better
targeted medicines.
Multidisciplinary Guidelines
Cross-cutting topics which do not fit
uniquely into one of the Quality,
Safety and Efficacy categories.
It includes the ICH medical
terminology (MedDRA), the Common
Technical Document (CTD) and the
development of Electronic Standards
for the Transfer of Regulatory
Information (ESTRI).

5. Validation
• Validation is the process of establishing
documentary evidence demonstrating that a
procedure, process, or activity carried out in
testing and then production maintains the
desired level of compliance at all stages.
• In the pharmaceutical industry, it is very important that in
addition to final testing and compliance of products, it is also
assured that the process will consistently produce the
expected results.

6. Analytical Method Validation
• is the process of demonstrating that an
analytical procedure is suitable for its
intended purpose.

7. Types of Analytical Procedures to be
Validated
 Identification tests
 Quantitative tests for impurities content
Limit tests for the control of impurities
Quantitative tests of the active moiety in
samples of drug substance or drug product or
other selected component(s) in the drug
product (Assay)

8. Validation Parameters as per ICH & USP
ICH
 Accuracy
 Precision
Repeatability
Intermediate Precision
Reproducibility
 Specificity
 Detection Limit
 Quantitation Limit
 Linearity
 Range
 Robustness
 System Suitability
USP
 Accuracy
 Precision
 Specificity
 Detection Limit
 Quantitation Limit
 Linearity & Range
 Ruggedness
 Robustness

9. Accuracy
 The accuracy of an analytical procedure expresses the
closeness of agreement between the value which is accepted
either as a conventional true value or an accepted reference
value and the value found.
 This is sometimes termed trueness.
Determination:-
1. Assay
Drug Substance
Drug Product
2. Impurities
Accuracy is calculated as % Recovery

10. Precision
 The precision of an analytical procedure expresses the
closeness of agreement (degree of scatter) between a series
of measurements obtained from multiple sampling of the
same homogeneous sample under the prescribed conditions.
 Precision may be considered at three levels:
1. Repeatability
2. Intermediate precision
3. Reproducibility
Determination:-by assaying of sufficient number of aliquots of
homogeneous samples

11. Repeatability:- Repeatability expresses the precision under the
same operating conditions over a short interval of time.
Repeatability is also termed intra-assay precision .
Intermediate precision:- Intermediate precision expresses
within-laboratories variations: different days, different
analysts, different equipment, etc.
Reproducibility:- Reproducibility expresses the precision
between laboratories (collaborative studies, usually applied to
standardization of methodology).

12. Specificity
 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.
 Lack of specificity of an individual analytical procedure may be
compensated by other supporting analytical procedure(s).
 This definition has the following implications:
– Identification: to ensure the identity of an analyte.
– 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.

13. Detection Limit
 The detection limit of an individual analytical procedure is
the lowest amount of analyte in a sample which can be
detected but not necessarily quantitated as an exact value.
 Formula:-
DL = 3.3 σ
S
where , σ = the standard deviation of the response
S = the slope of the calibration curve

14. Quantitation Limit
 The quantitation limit of an individual analytical procedure is
the lowest amount of analyte in a sample which can be
quantitatively determined with suitable precision and
accuracy.
 The quantitation limit is a parameter of quantitative assays for
low levels of compounds in sample matrices, and is used
particularly for the determination of impurities and/or
degradation products.
 Formula:-
QL = 10 σ
S
Where, σ = the standard deviation of the response
S = the slope of the calibration curve

15. Linearity
 The linearity of an analytical procedure is its ability (within a
given range) to obtain test results which are directly
proportional to the concentration (amount) of analyte in the
sample.
 For establishment of Linearity minimum of Five
concentrations normally used.
y = 0.0749x - 0.001
R² = 0.9998
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0 5 10 15
Abs
Conc
Abs
Linear (Abs)

16. Range
 The range of an analytical procedure is the interval between
the upper and lower concentration (amounts) of analyte in
the sample (including these concentrations) for which it has
been demonstrated that the analytical procedure has a
suitable level of precision, accuracy and linearity.
 Assay :- 80 - 120 %
 Impurity :- 50 - 120 %
 Content uniformity:- 70 – 130 %
 Dissolution :- 20 – 110 %

17. Ruggedness
• The ruggedness of the 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, analyst, instruments, lots of reagents, elapsed
assay time, assay temperatures or day.

18. Robustness
• 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.

19. System Suitability
• System suitability testing is an integral part of many analytical
procedures.
• The tests are based upon the concept that the equipment,
electronics, analytical operations and samples to be analysed
constitute an integral system that can be evaluated as such.
• System suitability test parameters to be established for a
particular procedure depend upon the type of procedure
being validated

20. Type of
analytical
procedure
characteristics
IDENTIFICATION
TESTING FOR
IMPURITIES
ASSAY
- dissolution
(measurement
only)
- content/potency
Performance
(Dissolution, Drug
Release )
Quantitative Limit
Accuracy - + - + -
Precision - + - + +
Repeatability - + - + -
Internal
Precision
- + (1) - + (1) -
Specificity (2) + + + + -
Detection Limit - - (3) + - -
Quantitation
Limit
- + - - -
Linearity - + - + -
Range - + - + -
- signifies that this characteristic is not normally evaluated
+ signifies that this characteristic is normally evaluated
(1) in cases where reproducibility (see glossary) has been performed, intermediate precision is not needed
(2) lack of specificity of one analytical procedure could be compensated by other supporting analytical procedure(s)

21. References
 Validation of Analytical Procedures: Text And Methodology Q2(r1);
International Conference On Harmonisation of Technical Requirements For
Registration Of Pharmaceuticals For Human Use Ich Harmonised Tripartite
; Current Step 4 Version Parent Guideline Dated 27 October 1994
(Complementary Guideline On Methodology Dated 6 November 1996
Incorporated In November 2005).
 General Chapter<1225>, Validation of Compendial Procedures, USP.