Validation and Control of Bioanalytical Methods

  • H. Thomas Karnes
  • Kumar A. Shah
Part of the Cancer Drug Discovery and Development book series (CDD&D)


The results of toxicokinetic, pharmacokinetic, and bioequivalence studies are used to make critical decisions regarding the safety and efficacy of anticancer drug substances. Therefore, measurement of anticancer drug concentrations in biological matrices is an important aspect in the development of these products. Such data are required by regulating agencies to support new drug applications as well as for line extensions and generic products of these drugs. It is therefore most essential to adequately characterize and fully validate the applied bioanalytical methods used in the determination of this class of compounds to ensure that they function in the manner in which they are intended. Since the release of the FDA prescribed Guidance for Industry in Bioanalytical Method Validation in May 2001, it is much clearer what is required for method validation. There are however a number of areas that are still not well developed in the FDA guidance, and the recently proposed draft European Medical Agency guidance addresses some of these. Apart from discussing acceptance criteria on the primary matrices required to determine bioanalytical assay suitability such as accuracy, precision, and selectivity, the draft guidance proposes additional criteria for other important aspects such stability tests, matrix effects, cross validation, and incurred sample reanalysis. The current chapter provides an overview of the current scientific approaches based on the literature while considering them in the context of these guidances in this highly regulated area.


Bioanalysis Method validation Acceptance criteria Quality control 


  1. 1.
    Shah VP (1987) Analytical methods used in bioavailability studies: a regulatory viewpoint. Clin Res Regul Aff 5:51–60CrossRefGoogle Scholar
  2. 2.
    Karnes HT, Shiu G, Shah VP (1991) Validation of bioanalytical methods. Pharm Res 8(4):421–426PubMedCrossRefGoogle Scholar
  3. 3.
    U.S. Department of Health & Human Services. Guidance for industry: bioanalytical method validation. FDA, CDER, CVM, May 2001 BPGoogle Scholar
  4. 4.
    Shah VP, Midha KK, Dighe S, McGilveray IJ, Skelly JP, Yacobi A, Layloff T, Viswanathan CT, Cook CE, McDowall RD, Pittman KA, Spector S (1992) Conference report. Analytical methods validation: bioavailability, bioequivalence and pharmacokinetic studies. Pharm Res 9(4):588–592CrossRefGoogle Scholar
  5. 5.
    U.S. Department of Health & Human Services. Guidance for industry. Bioanalytical methods validation for human studies. FDA, CDER, Dec. 1998 BPGoogle Scholar
  6. 6.
    Shah VP, Midha KK, Findlay JWA, Hill HM, Hulse JD, McGilveray J, McKay G, Miller KJ, Patnaik RN, Powell ML, Tonelli A, Viswanathan CT, Yacobi A (2000) Bioanalytical methods validation – a revisit with a decade of progress. Pharm Res 17:1551–1557PubMedCrossRefGoogle Scholar
  7. 7.
    Miller KJ, Bowsher RR, Celniker A, Gibbons J, Gupta S, Lee JW, Swanson SJ, Smith WC, Weiner RS (2001) Workshop on bioanalytical methods validation for macromolecules: summary report. Pharm Res 18(9):1373–1383PubMedCrossRefGoogle Scholar
  8. 8.
    European medicines agency: committee for medicinal products for human use – draft guideline on the validation of bioanalytical methods.
  9. 9.
    Curry SH, Whelpton R (1978) Statistics of drug analysis, and the role of internal standards. In: Reid E (ed) Blood drugs and other analytical challenges. Ellis Horwood, Chichester, pp 29–41Google Scholar
  10. 10.
    Haefelfinger P (1981) Limits of the internal standard technique in chromatography. J Chromatogr 218:73–81CrossRefGoogle Scholar
  11. 11.
    Figg WD, McLeod HL (2004) Handbook of anticancer pharmacokinetics and pharmacodynamics, 1st edn. Humana Press, Totowa, NJCrossRefGoogle Scholar
  12. 12.
    Massart DL, Andeginste BGM, Deming SN, Michotte Y, Kaufman L (1988) Chemometrics a textbook. Elsevier, New York, NYGoogle Scholar
  13. 13.
    Karnes HT, March C (1991) Calibration and Validation in Chromatographic Biopharmaceutical Analysis. J Pharm Biomed Anal 9:911PubMedCrossRefGoogle Scholar
  14. 14.
    Cassidy R, Janoski M (1992) Is your calibration curve linear? LC GC 10(9):692–696Google Scholar
  15. 15.
    Constanzer ML, Chavez CM, Matuszewski BK (1994) Picogram determination of finasteride in human plasma and semen by high-performance liquid chromatography with atmospheric-pressure chemical-ionization tandem mass spectrometry. J Chromatogr B 658:281–287CrossRefGoogle Scholar
  16. 16.
    Nelson MD, Dolan JW (2002) Ion suppression in LC-MS-MS – a case study. LC GC N Am 20(1):24–32Google Scholar
  17. 17.
    Hubaux A, Vos G (1970) Decision and detection limits for linear calibration curves. Anal Chem 42(8):849–855CrossRefGoogle Scholar
  18. 18.
    Long GL, Winefordner JD (1983) Limit of detection-A closer look at the IUPAC definition. Anal Chem 55:712A–722ACrossRefGoogle Scholar
  19. 19.
    Anderson RL (1987) Practical statistics for analytical chemists. Van Norstrand, New York, NYGoogle Scholar
  20. 20.
    Trissel LA (1983) Avoiding common flaws in stability and compatibility studies of injectable drugs. American Society of Hospital Pharmacists, Inc. 0002–9289.83/0701-1159900.50Google Scholar
  21. 21.
    Levey S, Jennings ER (1950) The use of control charts in the clinical laboratory. Am J Clin Pathol 20:1059–1066PubMedGoogle Scholar
  22. 22.
    Westgard JO, Barry PL (1986) Cost-effective quality control: managing the quality and productivity of analytical processes. AACC, Washington, DCGoogle Scholar
  23. 23.
    Causey AG, Hill HM, Phillips LJ (1990) Evaluation of criteria for the acceptance of bioanalytical data. J Pharm Biomed Anal 8:625–628PubMedCrossRefGoogle Scholar
  24. 24.
    Karnes HT, March C (1993) Precision accuracy and data acceptance criteria in biopharmaceutical analysis. Pharm Res 10:1420PubMedCrossRefGoogle Scholar
  25. 25.
    Kringle R, Khan-Malek R, Snikeris F, Munden P, Agut C, Bauer M (2001) A unified approach for design and analysis of transfer studies for analytical methods. Drug Inf J 35(4):1271–1288CrossRefGoogle Scholar
  26. 26.
    Dewé W, Govaerts B, Boulanger B, Rozet E, Chiap P, Hubert P (2007) Using total error as decision criterion in analytical method transfer. Chemometr Intell Lab Syst 85(2):262–268CrossRefGoogle Scholar
  27. 27.
    Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, Shah VP, Skelly JP, Swann PG, Weiner R (2007) Workshop/conference report—quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. AAPS J 9(1):E30–E42CrossRefGoogle Scholar
  28. 28.
    Fast DM, Kelley M, Viswanathan CT, O’Shaughnessy J, King SP, Chaudhary A, Weiner R, DeStefano AJ, Tang D (2009) Workshop report and follow-up—AAPS workshop on current topics in GLP bioanalysis: assay reproducibility for incurred samples—implications of crystal city recommendations. AAPS J 11(2):238–241PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of PharmaceuticsVirginia Commonwealth University School of PharmacyRichmondUSA
  2. 2.Research Scientist R&D, Chromatographic Sciences Department, PPDRichmondUSA

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