This presentation gives a summary of the New Draft WHO Guidance on Analytical Method Validation.

1. This presentation is compiled by “ Drug Regulations”
a non profit organization which provides free online
resource to the Pharmaceutical Professional.
Visit http://www.drugregulations.org for latest
information from the world of Pharmaceuticals.

2. This presentation is compiled from freely available resource
like the website of WHO specifically WHO Draft Guidance
Titled
 GUIDELINES ON VALIDATION – APPENDIX 4 ANALYTICAL
METHOD VALIDATION
“Drug Regulations” is a non profit organization which
provides free online resource to the Pharmaceutical
Professional.
Visit Our Website GMP Training
for latest information from the world of Pharmaceuticals.
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3.  This presentation is based on WHO
Draft Guidance titled
 GUIDELINES ON VALIDATION –
APPENDIX 4 ANALYTICAL METHOD
VALIDATION
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4.  The need for revision of earlier
Supplementary guidelines on good
manufacturing practices: validation was
identified by the Prequalification of Medicines
Programme
 A draft document was circulated for comment
in early 2013.
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5.  The focus of the revision was the Appendix on non-
sterile process validation (Appendix 7), which had been
revised and was adopted by the Committee at its forty-
ninth meeting in October 2014.
◦ See our presentation here
 Modified Appendix 4 on Analytical Method Validation is
now published in working document QAS/16.671.
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6.  This guidance covers following
◦ Principle
◦ General
◦ Pharmacopoeial Methods
◦ Non Pharmacopoeial Methods
◦ Method Validation
◦ Method Verification
◦ Method Transfer
◦ Revalidation
◦ Characteristics of Analytical Methods
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7.  This guidance presents information on
the characteristics that should be
considered during Analytical method
Validation.
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8.  Approaches other than that specified in
this guidance can be adopted.
 Manufacturers should choose validation
protocols and and procedures most
suitable for testing their products.
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9.  It is essential to demonstrate that the
Analytical Procedure is suitable for
intended use.
 All analytical methods should be
validated.
◦ Whether they are stability indicating or not
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10.  Validation of Analytical method should be performed by
the R & D
 There after the method can be transferred to Quality
Control.
 Follow recommendations provided in Good Laboratory
practices guidance
 Also follow the recommendations given in Transfer of
Technology Guidance.
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11.  Have specifications for both materials
and products.
 The tests to be performed should be
described in the documentation on
Standard Test Methods.
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12.  Pharmacopeial or Non Pharmacopeial
Methods May be Used.
 Well Characterized reference materials
with documented purity should be
used in analysis.
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13.  Most common Analytical procedures Include
◦ Identification Tests
◦ Assay of Drug Substance
◦ Assay of Pharmaceutical Products
◦ Quantitative tests for content of Uniformities
◦ Limit Test for Impurities
◦ Dissolution Testing
◦ Determination of Particle Size.
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14.  The Results of Analytical Procedures should be
◦ Legible
◦ Contemporaneous
◦ Original
◦ Reliable
◦ Reproducible
 All Results should be archived for an appropriate period of
time.
 They should be in compliance with NRA requirements
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15.  The procedure should become part of
a continuous verification procedure
 Demonstrate that it meets predefined
criteria over the life of the procedure.
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16.  Perform following at intervals to
ensure that the method is appropriate
for its intended use.
◦ Trend Analysis
◦ Risk Assessment
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17.  Manage changes to methods as per COC procedure
 Consider following when methods are transferred from
one lab to another or when major pieces of analytical
equipment are changed:
◦ Variability of Reference materials.
◦ Changes in process of synthesis
◦ Changes in composition of the finished product
◦ Changes in Analytical Procedure
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18. The changes referred earlier should be
◦ Understood
◦ Controlled
◦ Where possible reduced.
 Consider Verification or Revalidation
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19.  Perform risk assessment for such
changes
 Scope of verification & degree of
revalidation depend on
◦ Nature of changes
◦ Out come of risk assessment
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20.  Analyst should be appropriately qualified.
 Qualification should be documented
 For certain tests conduct “Analyst Proficiency”
 Data for method validation and verification should be covered
by GxP
 Follow good data and record management.
 Associated metadata should also be retained and be
subjected to good data and record management practices.
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21.  Use of computerized systems should comply with
the principles given in Guidance for Computerized
Systems Validation.
 Pay Adequate attention to sample preparation.
 This step should be described in as much detail as
possible.
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22.  Attention should be paid to factors such as :
◦ Sonification Time
◦ Sonification Bath Temperature
◦ Mixing
◦ Demixing where such is known to happen
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23.  Document & report all failures during method validation.
 Also document how these failures were overcome
 It is not acceptable to give only passing results.
 This will give biased information about reliability of the
method.
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24.  When such method are used there should be evidence that such
methods are suitable for routine laboratory use.
 i.e. The methods should be verified.
 When such methods are used for determination of content or
impurities , the specificity of the methods should be demonstrated.
 i.e. Specificity with respect to the substance under consideration –
no placebo interference.
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25.  All Non Pharmacopoeial Methods should be
appropriately validated.
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26.  Perform validation as per a protocol.
 Protocol should include procedure and acceptance criteria for all
characteristics.
 Results should be documented in the validation report.
 Justification should be provided if Non Pharmacopoeial methods are
used when Pharmacopoeial are available.
 Justification should include data for comparison with Pharmacopoeil
and other methods.
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27.  Standard Test methods should be described in details.
 They should provide sufficient information to allow a properly
trained analyst to perform analysis in reliable manner.
 As a minimum, the description should include
chromatographic conditions, reagent needed, reference
standards, formulae for calculation of results and system
suitability tests.
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28.  Method verification consists of partial velidation.
 It should be performed for a validated method under following
conditions:
◦ When an already validated method is used on a product for the first time.
 Change of active ingredient supplier, change in method of synthesis,
reformulation of a drug product
◦ When an already validated method is used in a laboratory for the first
time.
 In some cases method transfer may be preferred.
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29.  May include only the validation characteristics of relevance to
the particular change.
◦ In case of change in API supplier the expected change would be
impurity profile or solubility of the API.
◦ Therefore for an Related substances method there should be an
appropriate verification that the method is able to detect and
quantitate all potential impurities including the late eluting ones.
◦ Specificity should be amongst the tests considered.
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30.  Method Verification is suitable
in lieu of method validation for
Pharmacopoeial methods.
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31.  Methods should be maintained in validated state over the
life of the method.
 Perform Revalidation when there are following changes:
◦ Changes to the Mobile Phase( Refer Pharmacopoeias for acceptance limits
beyond which revalidation must be performed.
◦ Changes to the column
◦ Changes to the temperature of the column
◦ Changes to the concentration/composition of the sample and standards
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32.  Perform Revalidation when
there are following changes:
◦ Changes to the detector
 Change in detector type
 Wavelength of detection
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33.  Perform Revalidation when there are
following changes:
◦ In cases of repeated system suitability failures
◦ In case of doubtful results
◦ In such cases perform an investigation to determine root
cause.
◦ Then make appropriate changes and revalidate the methods
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34.  Perform revalidation according to a
period that is scientifically justifiable.
 It is acceptable to include only the
validation characteristics of relevance to
the particular change and methods
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35.  Establish Documented evidence that
◦ a method has equivalence performance
when used in a laboratory different from
that where it was originally validated.
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36.  Generally it should be performed
◦ By comparing a set of results obtained by
an analyst in one laboratory
◦ to that obtained by another analyst at
the laboratory to which the method is
being transferred.
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37.  Two set of results should be
statistically compared
 The differences between the two
sets of test results should be
within an acceptable range.
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38.  Method transfer should be performed before
testing of samples for obtaining critical data.
◦ For Dossier
◦ Process Validation
◦ Stability Studies
◦ For Routine use
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39.  Follow a predetermined protocol which should include:
◦ A title
◦ Objective
◦ Scope
◦ Responsibilities of the sending unit & the receiving unit
◦ A specification of materials and methods
◦ Experimental design
◦ Acceptance criteria
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40.  Follow a predetermined protocol which should include:
◦ Documentation: Information to be supplied with results, forms
etc.
◦ Deviation handling procedure
◦ References
◦ Details of Reference samples, starting materials, intermediates
and finished products
 Protocol should be authorized and dated
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41.  Independent Testing by a separate entity: National Quality
Control Testing Laboratory.
 This may not be considered an obligation.
 This may be considered an optional step when
encountering difficulties in applying a particular method.
◦ Refer WHO Guidance on technology Transfer
 (See our Presentation)
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42.  Consider following Characteristics while validation:
◦ Specificity
◦ Linearity
◦ Range
◦ Accuracy
◦ Precision
◦ Detection Limit
◦ Quantitation Limit
◦ Robustness
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43.  Accuracy
◦ Degree of agreement of Test Results with the True Value
◦ Closeness of the results obtained by the procedure to the true
value.
◦ Generally established on samples of the material to be
examined that have been prepared to quantitative accuracy.
◦ Should be established across the range specified for the
analytical procedure.
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44. Accuracy
◦ Acceptable to use a spiked placebo
where a known quantity or
concentration of a reference material
is used.
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45.  Precision
◦ Degree of agreement among individual results.
◦ Apply complete procedure repeatedly to separate
identical samples drawn from the same homogenous
batch of materials.
◦ Measured by the scatter of individual results from the
mean.
◦ Expressed as the Relative Standard Deviation-RSD
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46.  Repeatability
◦ Determine using a minimum of nine
determinations covering the specified range for
the procedure.
◦ Three concentration/three replicates each or
◦ A minimum of six determinations at 100 % of the
test concentrations.
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47.  Intermediate Precision
◦ This expresses within laboratory variations
 Different days
 Different analysts
 Different equipment
◦ If reproducibility is assessed , a measure of
intermediate precision is not required.
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48. Reproducibility
◦ Expresses precision between
laboratories
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49.  Robustness( or Ruggedness)
◦ Ability of the procedure to provide
analytical results of acceptable accuracy
and precision under a variety of
conditions.
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50.  Robustness( or Ruggedness)
◦ Results from separate samples are influenced by
changes in the operational or environmental
conditions.
◦ Robustness should be considered during the
development phase
◦ Should show the reliability of an analysis when
deliberate variations are made in method parameters.
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51.  Robustness( or Ruggedness)
◦ Verification of stability of analytical solutions is of importance.
◦ Other characteristics of robustness should include extraction
time.
◦ In case of Liquid chromatography robustness testing may also
include
 Verification of the impact of changes in pH, temperature and
flow rate.
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52. ◦ Factors that can have an effect on robustness when performing
chromatographic analysis include:
 Stability of standard & test solutions
 Reagents
 Different columns-different lots/suppliers
 Extraction time
 Variations of pH of mobile phase
 Variations in mobile phase compositions
 Temperature
 Flow Rate
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53.  Linearity
◦ Indicates ability to produce results that are directly
proportional to the concentration of the solution.
◦ Use samples in which the analyte concentrations span
the claimed range of the procedure.
◦ In case of a linear relationship use appropriate
statistical test to show linearity.
◦ A minimum of 5 concentrations should be used.
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54.  Range
◦ Expression of the lowest and highest level
of analyte that have been demonstrated to
determinable for the product.
◦ Specified range is normally derived form
linearity study.
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55.  Specificity ( selectivity)
◦ Ability to measure unequivocally the desired analyte
in the presence of components such as excipients and
impurities that may also be expected to be present.
◦ An investigation of specificity should be performed
during the validation of identification tests, the
determination of impurities and assay.
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56.  Detection Limit ( Limit of Detection)
◦ Smallest quantity of analyte that can be detected and not necessarily
determined in a quantitative fashion.
◦ Approaches may include instrumental or non-instrumental procedures
◦ Could include those based on
 Visual evaluation
 Signal to noise
 Standard deviation of the response and the slope
 Standard deviation of the blank
 Calibration curve
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57.  Quantitation Limit ( Limit of Quantitation)
◦ Lowest concentration in a sample that may be determined with acceptable
accuracy and precision.
◦ Approaches may include instrumental or non instrumental procedures
◦ Could include those based on
 Visual evaluation
 Signal to noise
 Standard deviation of the response and the slope
 Standard deviation of the blank
 Calibration curve
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58. 04-09-2016 58Visit Our Website GMP Training

59.  Use appropriate statistical analysis to
evaluate validation characteristic against
predetermined acceptance criteria.
 Use appropriately validated software
 Consider appropriate number of samples to
provide adequate statistical power and range.
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60.  Integral part of analytical testing procedure.
 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.
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61.  SS parameters that need to be
established for a particular
procedure depend on the
◦ Type of procedure being evaluated
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62.  Suitability of the entire system
should be confirmed prior to and
during method validation tests.
 Similarly this should be done
during sample analysis.
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63.  SS runs should include only established
standards or reference material of known
concentrations.
 This provides appropriate comparator for
the potential variability of the
instrument.
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64.  When a sample is used for system
suitability written procedures should be
established.
 Results of all such trial runs should be
included in the results and data review
process.
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65.  A sample can be used only if it is well
characterized material.
 Characterization in such cases should
be performed prior to the use of this
sample as part of SS testing.
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66.  The sample material or product under
test should not be used for trial run
purposes or to evaluate the SS.
 Refer WHO guidelines on data and
record management.
◦ See our presentation here.
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67. This presentation is compiled from freely available resource
like the website of WHO specifically WHO Draft Guidance
Titled
 GUIDELINES ON VALIDATION – APPENDIX 4 ANALYTICAL
METHOD VALIDATION
“Drug Regulations” is a non profit organization which
provides free online resource to the Pharmaceutical
Professional.
Visit Our Website GMP Training
for latest information from the world of Pharmaceuticals.
04-09-2016 67
Visit Our WEBSITE GMP Training
http://fdagmp.blogspot.in/