This document discusses validation of analytical methods. It defines validation as establishing evidence that a process will consistently produce results meeting specifications. Validation guidelines include ICH Q2A, Q2B, FDA guidance, and pharmacopoeias. Key validation characteristics covered are specificity, linearity, range, accuracy, precision, detection/quantitation limits, robustness, and system suitability testing. Equipment validation involves design qualification, installation qualification, operational qualification, performance qualification, and process qualification in three phases: pre-validation, process validation, and validation maintenance.

1. VALIDATION
OF
ANALYTICAL
METHOD

2. DEFINITIONS
FDA-GUIDELINES
Validation is establishing documented evidence which provides a high degree of
assurance that a specific process will consistently produce a product meeting its
pre-determined specifications and quality attributes
EU-GUIDELINES
Action of proving, in accordance with GMP-principles that any
procedure, process, equipment, material, activity or system actually leads to the
expected results
More specific:
As per ICH guideline
(Topic Q2B, March 1995 )
“Methods validation is the process of demonstrating that analytical procedures
are suitable for their intended use”

3. VALIDATION GUIDELINES
• ICH Q2A:Text on validation of analytical procedures:
Definitions and terminology (March 1995)
• ICH Q2B : Validation of analytical procedures: Methodology (June 1997)
• FDA : Guidance for Industry: Analytical procedures and methods validation
• Pharmacopoeias :USP and European Pharmacopoeia

4. TYPES OF VALIDATION

5. WHAT ASPECTS TO COVER?
• Specificity
• Linearity
• Range
• Accuracy
• Precision
• Detection limit
• Quantitation limit
• Robustness
• System Suitability Test

6. SPECIFICITY
• Definition:
Ability to assess unequivocally the analyte in the
presence of of components which may be expected
to be present (impurities, degradants, matrix) .
Aspects:
- Identification
- Purity tests
- Assay (Content/potency)

7. LINEARITY
• Definition:
Ability (within a specified range) to obtain test results which are
directly proportional to the concentration of analyte in the sample
• Aspects:
- Test across the range (at least 5 concentrations) (80% to 120
%)
• - Evaluate linearity by visual inspection of the plot and by
statistical techniques
• - Calculate corr. coefficient, y-intercept, slope and res. sum of
squares (r > 0.99 )

8. RANGE
• Definition:
Interval between upper and lower concentration of the analyte in the
sample for which it has been demonstrated that the procedure has a
suitable level of precision, accuracy and linearity .
• Aspects:
- Defined from linearity study
• - Depends on the application of the method (assay, dissolution test,
content uniformity)

9. ACCURACY
• Definition:
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
Methods:
Drug substance
• use of reference standard with known purity
• comparison with independent, well-characterised procedure
• may be inferred once precision, linearity and specificity are established.
Drug product
• spiking of placebo mixture
• addition of analyte to ‘active’ material
• comparison of results obtained with independent, well-characterised procedure
• may be inferred once precision, linearity and specificity are established .
Recommended Data :
• Assessed by 9 determinations over a minimum of 3 concentration levels covering
the specified range
• To be reported as percent recovery.

10. ACCURACY

11. PRECISION
• Definition
Closeness of agreement (‘scatter’) between a series of
measurements obtained from multiple sampling of the same
homogeneous sample.
Aspects
• Repeatability
• Intermediate precision
• Reproducibility

12. Precision - Repeatability
Precision under the same operating conditions over a short interval of
time.
Precision - Intermediate precision
Expresses within laboratory variations.
Precision - Reproducibility
Precision between laboratories

13. DETECTION LIMIT
or
LIMIT OF DETECTION (LOD)
• Definition
Lowest amount of an analyte in a sample which can be detected but not
necessarily quantitated.
Method
- Based on visual evaluation
- Based on signal-to-noise ratio (3:1)
- Based on st.dev. (SD) of response and slope (DL=3.3xSD/S) - Report results
and method of choice

14. QUANTITATION LIMIT
or
LIMIT OF QUANTITATION (LOQ)
• Definition
• Lowest amount of an analyte in a sample which can be
quantitatively determined with a suitable precision and
accuracy
• Method
- Based on visual evaluation
- Based on signal-to-noise ratio (10:1)
- Based on st.dev. (SD) of response and slope (DL=10xSD/S) -
Report results and method of choice

15. ROBUSTNESS
• Definition
Measure of the capacity of a method to remain unaffected by small variations in
method parameters.
Aspects
- To be considered during development
- To be used for establishment of system suitability criteria
- Include testing of stability of solutions
- To be tested by introducing small variations in method parameters

16. SYSTEM SUITABILITY TEST
• Definition
Set of parameters and criteria there off to ensure the system is working properly.
Aspects
• For chromatographic methods: tailing factor, rel. retention times, resolution
factor, rel. st. deviation, number of theoretical plates
• - To be checked before start of run and to be verified afterwards - Described
in Pharmacopoeias

17. Recommended Validation characteristics of various Types of Tests

18. EQUIPMENT VALIDATION
• Definition:
1. Equipment validation is established documented set up
that proves any equipment works correctly and leads to
accepted and accurate results .
2. The process of equipment validation is based on the principle that
equipment must be designed, constructed, maintained, and adapted to
perform the operations which are to be carried out .

19. TYPES OF EQUIPMENT VALIDATION
The process of equipment validation is not a single step activity that it has different
phases which have further subsections or steps, these are as follow
1. Design qualification
2. Installation qualification
3. Operational qualification
4. Performance qualification
5. Process qualification
PHASES OF EQUIPMENT VALIDATION
The process of equipment validation is mainly divided into three phases
1. Phase – 1: Pre-validation phase.
2. Phase – 2: Process validation phase.
3. Phase – 3: Validation maintenance phase.

20. CLASSIFICATION

21. PRE-VALIDATION PHASE
Design Qualification (DQ) :
• It is a documented verification of design of the equipment and manufacturing
facilities
• The purpose of (DQ) is to make sure that all the requirements for the
systems should clearly defined at the start
• It defines the functional and operational specifications of the instrument
• The accomplishment of documented qualification must verify that the given
design will follow :
1. User requirement specification ( URS)
2. Functional specification (FS)
3. Tender specification and drawing
4. Purchase specification Vendor qualification

22. PRE-VALIDATION PHASE
• INSTALLATION QUALIFICATIONS (IQ):
• It confirms that the précised equipment has been received and installed as
per target and agreement in exact design or format in the undamaged form.
• It is documental verification of that the equipment has been installed and
calibrated appropriately
• As per the manufacture’s recommendations for installation, the working sites
working environmental conditions are documented and confirmed that they
are suitable for the operation of the instrument.
The documentation of installation includes:
1. Details of supplier and manufacture.
2. Equipment name, colour, model and serial number
3. Date of installation and calibration.

23. PROCESS VALIDATION PHASE
1.Operational Qualifications:
1. Operational qualification ensures that installed equipment/instrument will
function perfectly according to its operation specification in the mention
environmental conditions.
2. It also checks that the equipment function perfectly to meet pre-assigned
performance criteria and ensure how the testing results are recorded.
3. operational qualification gave high degree of assurance that the equipment
functionally verifies compliance of manufactures specifications.
Documentation for operational validation includes:
1. Finalized and approved operations (functions testing)
2. Certified calibrations
3. System stability test results
4. Applications of S.O.P.s

24. PROCESS VALIDATION PHASE
• Performance qualification :
• It Ensures that the equipment consistently performs functions according to
the mentioned specification
• It is a documented verification process which verifies that all aspects of
facility, utility, and performance of equipment meeting pre-assigned
acceptance criteria
Documentation for performance validation includes
1. Performance qualification report
2. Process stability testing reports (long-term productivity)
3. Acceptance of the product record (costumers reviews)
4. Actual product and process parameters documentations.
• Routinely performed test results documentation

25. RE-VALIDATION
• The performance of re-validation is done when the operating equipment and
system have been modified in some ways due to any reason
• Revalidation of the equipment is very helpful in maintaining the validation
status of the equipment and entire system which work as a unit
• Re-validation is further divided as follow:
• Periodic/scheduled re-validation
• Re-validation after change/modifications
Periodic re-validation :
The re-validation process which carried out in pharmaceutical industry at
periodic intervals and it is mandatory especially when the company made any
change in the formulas, procedures, manufacturing systems, packaging, and
support system such as electricity/ power supply, water supply, and steam

26. RE-VALIDATION
The following is the changes for that re-validation:
• Change in raw material.
• Change in manufacturing process.
• Change in equipment/system.
• Change in supporting systems.
• Change in packaging materials.

27. VALIDATION MAINTENANCE PHASE
• MAINTENANCE QUALIFICATION (MQ) :
• Maintenance qualification will review and verify the acceptability of the
maintenance controls to confirm the equipment/ system integrity
• It is a periodic process which ensures that the equipment should not affect
the safety, quality, and strength, identity of the manufactured product either
through its
contamination or structure.
• The process of maintenance qualification includes routine servicing and
necessary repairs.
• The documentation for maintenance qualification includes:
 outline services records
 Maintenance contracts details List of authorized services engineers

28. APPLICATION OF EQUIPMENT VALIDATION
• Validation of equipment reduces costs by reducing rejects, reworks, and
downtime.
• High rate of customer satisfaction.
• Analytical tests methods and calibrations are proceeded
• It also reduces testing in in-process and final product.
• Also improve employee’s awareness
• Make maintenance of equipment easier.
• Give more rapid and reliable start-up for new equipment’s.
• Help in development of validation master plan for the facility