1. Evolution & Changes in Monograph/Guidelines:
Issues in Analytical Development
Dr. Bhaswat S. Chakraborty
Senior Vice President, R&D, Cadila Pharmaceuticals
Waters Technology Seminar, Hyderabad, Dec. 10, 2008

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Hood et al. Nature Biotechnology 22, 1215 - 1217 (2004)

4. Main Changes in Guidelines and Monographs
 From an analytical perspective, the main changes have been
 More and more drugs are now covered by pharmacopeial monographs
 Assay and RSs better defined
 Compendial methods exist
 Better understanding the sources of impurities
 FDA guidances on method validation, impurity profiling are in place and
harmonized
 Q1, Q2, Q3, Q6A, FDA Analytical guidelines
 USP 31 Ch <1225>, <1226>
 Genotoxic impurities guidelines
 Association (Mfg., Medical, Scientific) guidelines
 Erudite papers
 Although, most of the changes have been an improvement, the
volume and complexity have also increased tremendously

5. Guidelines and Pharmacopeial Monographs
 Documents of excellence BUT….
 Scientific (public or private standards) but treated as
public “must do” regulations
 Regulatory guidelines
 USP, NF, IP, BP
 Often taken very seriously (rather rigidly) by reviewers
and expert audit inspectors
 Open to interpretations
 Often encourage inclusion of “nice to know” in the same
breadth as the “must know” information as long as it
makes scientific sense
 Can be idealistic rather than pragmatic

6. Today
 For a moment, understand the development of guidelines and
monographs and why they can be sometimes complex and
over-demanding
 Current validation
 The thrust on keeping the target error in control
 Special stress on impurrities
 Unknown, toxic
 When do we
 Method transfer
 Method verification
 Full (de novo) method validation
 Examples
 Write to FDA and consult Experts?
Discussion

7. Regulatory agencies appoint
Internal Guideline Committees
Guidelines Development
or Working Groups or Expert
Advisory Committees They deliberate
and come up with
draft guidelines
Acceptance by the
Ministry/Agency
& the Stakeholders
Draft guidelines are commented upon by (may
be 1-4 rounds)
Experts
Industry
Provincial Formularies, FDAs or
Regulatory governments
Impact Other stakeholders like professional
Analysis associations

8. Monographing Process
Source: R. Williams & Expert Project Team 4, J Pharm Biomed Anal (2006), 40, 3-15

9. ICH Method Validation Parameters
 LOD
 LOQ
 Precision
 Accuracy
 Specificity
 Linearity
 Assay range
 Robustness
 System suitability

10. Data Elements Required for Validation
Category II
Analytical
Performance Category I Quantitative Limit Tests Category III Category IV
Characteristics
Accuracy Yes Yes * * No
Precision Yes Yes No Yes No
Specificity Yes Yes Yes * Yes
Detection Limit No No Yes * No
Quantitation
No Yes No * No
Limit
Linearity Yes Yes No * No
Range Yes Yes * * No
*May be required, depending on the nature of the specific test.

11. Data Elements Required for Validation
 Category I – Analytical procedures for quantitation of major components
of bulk drug substances or active ingredients(including preservatives) in
finished pharmaceuticals products.
 Category II – Analytical procedures for determination of impurities in bulk
drug substances or degradation compounds in finished pharmaceuticals
products. These procedures include quantitative as says and limit tests.
 Category III – Analytical procedures for determination of performance
characteristics (e.g., dissolution, drug release).
 Category IV – Identification tests.
 For each category, different analytical information is needed. Listed in
Table 2 are data elements that are normally required for each of these
categories.

12. More & More Reflection of ICH in USP
 During the 2000–2005 cycle, USP created a Guideline for
Submitting Requests for Revision to USP–NF
 The Guideline provides instructions to Sponsors intending to
submit Requests for Revision and harmonizes many elements
of the USP monograph with the ICH Quality approaches
 USP expects to revise this document continuously
 For selected candidate reference materials, USP will add a
section that provides a protocol with study design and analysis
approaches
 This protocol will focus initially on small molecule ingredient
and impurity candidate materials and can be expanded
subsequently for other candidate materials as needed
Source: R. Williams & Expert Project Team 4, J Pharm Biomed Anal (2006), 40, 3-15

13. Analytical Procedures & Validations
 Regulatory or Compendial
 Analytical procedures in Pharmacopeia/Formulary recognized legally
 Alternative
 Equal to or better than the regulatory analytical procedure.
 Provide a rationale for its inclusion and identify its use
 e.g., release, stability testing
 validation data, and comparative data to the regulatory analytical procedure
 Stability-indicating assay
 A validated procedure that can analyse changes with time in the pertinent
properties of the drug substance and drug product
 Validations
 Full validation
 Revalidation
 Verification or Partial validation
 System suitability
 Method transfer

14. Full Validation and Revalidation
 ICH Method Validation Parameters
 LOD
 LOQ
 Precision
 Accuracy
 Specificity
 Linearity
 Assay range
 Robustness
 System suitability and
 Stability of samples over period of analysis
 Information from stress studies
 Impurities labeled with their names and location identifiers
 ….next slide

15. Full Validation
(Prednisolone)
Source: Gorog et al, J Pharm Biomed Anal (1998), 18, 511-525

16. System Suitability
 Tailing factor
 _ Relative retention
 _ Resolution
 _ Relative standard deviation (RSD)
 _ Capacity factor
 _ Number of theoretical plates

18. Impurities
 APIs
 [mfg. & storage] Process & drug related organic impurities
 Inorganic impurities
 Residual solvents
 Formulations
 Those forming during formulation
 Method related
 Environment related
 Dosage form factor related
 Those forming on aging
 Ingredient interactions
 Functional group related degradations
 Hydrolysis,
 Oxidative
 Photolytic
 Decarboxylation

19. Impurities Profiling
 Pharmacopeial impurities are controlled through the
specifications of
 Related substances
 Chromatographic purity tests
 System suitability
 Response factors
 Does not consider differences in route of synthesis
 ICH overcomes this
 Stability (Q1)
 Analytical validation (Q2)
 Impurities (Q3)
 Test Procedures and Acceptance Criteria for New Drug Substances and
New Drug Products (Q6A)

20. Ideal Control of Impurities
 Identifies all impurities >0.1%
 Even <0.1% (sometimes in low ppm) if unusually potent or toxic
 In any case, Qualification threshold must well defined
 Official reference standards for all impurities are available
 Route of synthesis is public or known to regulatory agencies
 Both process-related and degradation impurities are identified and
quantitated when required
 When changes in monograph impurities are published, clear instructions are
given whether full, partial or system suitability validations are required
 Commitment by manufacturers, regulators, Pharmacopeias to stop
counterfeits

21. ICH Thresholds for Impurity Identification &
Quantification (Finished Products)
Dose Identification (%) Quantification (%)
<1 mg 1.0 1.0
1-10 mg 0.5 1.0
10-100 mg 0.2 0.5
100 mg – 2 g 0.1 0.2
>2 g 0.1 0.1

22. Yet there are Many issues
 And the common ones are:
 If the method is compendial, do I get a method transfer only with
a DMF sourcing? or
 Validate partially?
 Validate fully?
 Should my method cover all known and unknown impurities?
 What should be range and LOQ?
 Should LOQ be a part of impurity specifications?

23. Process
Impurities
(Trimethoprim)
Source: Rao & Nagaraju, J Pharm Biomed Anal (2003), 33, 335-377

24. Degradation
Products
(Ramipril)
Source: Belal et al, J Pharm Biomed Anal (2003), 24, 335-342

25. Photodegradation
(Trifluperazine)
Source: Abdel-Moety, J Pharm Biomed Anal (1996), 14, 1639-1644

26. Source: J. Ermer, J Pharm Biomed Anal (1998), 18, 707-714

27. When do You do a Full Validation?

28. System Suitability Tests (Fenofibrate)
System Suitability. Six 5 ml aliquots of the system suitability solution were injected into the
system. The system was deemed to be suitable if the efficiency of the column, calculated using the
fenofibrate peak, was not less than 7000 plates, the resolution between compound V and fenofibrate was
not less than 20, the retention time of fenofibrate was about 7.3 min, the relative retention time of
compound V about 0.26, and the R.S.D. of the peak response from fenofibrate was not more than 5.0%.
Source: Lacroix et al, J Pharm Biomed Anal (1998), 18, 383-402

29. Source: Lacroix et al, J Pharm Biomed Anal (1998), 18, 383-402

30. Validation vs. Qualification vs. Method
Transfer
 Method Validation (Full Validation)
 Assess all appropriate validation characteristics
 Pre-defined acceptance criteria
 Follows formal validation protocol/sign off by the QU
 ICH Guideline:
 Q2 (R1): Validation of Analytical Procedures: Text and Methodology
 Method Qualification (Partial validation, suitable for
it’s intended purpose)
 Assesses a critical subset of validation characteristics
 No pre-defined acceptance criteria
 No official guidelines
 Suitable for early IND studies, characterization assays

31. Validation vs. Qualification vs. Method
Transfer
 Method Validation (Full Validation)
 Assess all appropriate validation characteristics
 Pre-defined acceptance criteria
 Follows formal validation protocol/sign off by the QU
 ICH Guideline:
 Q2 (R1): Validation of Analytical Procedures: Text and Methodology
 Method Qualification (Partial validation, suitable for
it’s intended purpose)
 Assesses a critical subset of validation characteristics
 No pre-defined acceptance criteria
 No official guidelines
 Suitable for early IND studies, characterization assays

32. Validation vs. Qualification vs. Method
Transfer
 Method Transfer
 Transfer of validated analytical procedures to a new laboratory
 Assesses a subset of validation characteristics
 Pre-defined acceptance criteria
 No official guidelines

33. Method Transfer
 Transfer of validated analytical methods from originating
laboratory to secondary laboratory
 To ensure comparability in the validation characteristics between laboratories
 To prevent and/or detect changes in data trend
 Assess a subset of validation parameters
 Using Equivalence Testing
 Precision and Accuracy
 Using Key attributes
 Precision
 LOD/LOQ
 Accuracy
 Identity
 Linearity
 It may be useful to analyze historical data from the originating
site to identify the greatest causes of variance in an assay to
improve transfer success

34. Method Transfer
 A recommended approach is to use equivalence
testing as the statistical approach
 Assumes not equivalent as the default hypothesis
 Concludes equivalent with enough evidence
 Does not penalize large sample size
 Need a predefined meaningful allowable difference
 Traditional hypothesis testing assumes equivalence as
default
 Rewards assays with high variability
 Penalize large sample sizes, tiny differences will be significant if sample size
is large enough
 Not a reasonable approach

35. Prior to Formal Method Transfer
 Receiving laboratory should perform the
method
 Helps to determine where there are differences
and gaps in documentation
 Lack of detailed test method instructions
 Assay Conditions
 Calculations
 System Suitability
 Differences with instrumentation or reagents

36. Prior to Formal Method Transfer
 Training of Personnel
 Review of relevant SOPs
 Observation of test procedure
 Performing test procedure
 Helpful to include development, qualification and
validation reports to recipient laboratory

37. Method Transfer
 Typical Transfer should include:
 More than one lot of material
 Reference Standards
 Samples at extremes of the established acceptable limits
 Stress samples
 One laboratory, typically the originating laboratory, will
prepare initial samples to be tested at both laboratories.
 One analyst at the originating laboratory and multiple
analysts in the receiving laboratory

38. Method Transfer Protocol
 Test Method
 Parameters being assessed
 Precision, specificity, etc.
 Sample Preparation/Reagent
 Performance Parameters
 Different analyst on different days
 Pre-defined Acceptance Criteria
 Statistical Analysis
 Sign-off by Quality Assurance Unit

39. Successful Method Transfers
 Pre-defined acceptance criteria using an appropriate
statistical approach.
 Prior to transfer, perform assay, train personnel and
determine if differences in equipment and reagents effect
the assay results.
 Test more than one lot of material.

40. Successful Method Transfers
 Include in the Regulatory Submission:
 Transfer Protocol
 Final Transfer Study Report
 Include representative and/or full data sets
 Deviations
 Statistical analysis
 Conclusions

41. Thanks