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Analysis of Imprecision in Incurred Sample Reanalysis for Small Molecules

The AAPS Journal volume 17, pages 206–215 (2015)Cite this article

Abstract

Over the years, incurred sample (IS) reanalysis (ISR) has become a tool to confirm the reliability of bioanalytical measurements. The recommendation for ISR acceptance criterion for small molecules is at least 67% of ISR samples that have reanalyzed concentrations within 20% of their original concentrations when normalized to their means. To understand the relevance of the ISR acceptance criterion and sample size requirements, simulated ISR studies evaluated the probability of ISR studies passing the acceptance criterion (ISR pass rate) as a function of IS imprecision and sample size. When IS imprecision (percent coefficient of variation: %CV) is low (≤10 or 1–10% CV), high ISR pass rate (≥99%) is attained with <50 samples. At intermediate IS imprecision (e.g., 12% CV or 7–12% CV range), 80–160 samples are required for a high ISR pass rate. When IS imprecision is at the higher end of the acceptance limit, ISR pass rate decreases significantly, and increasing sample size fails to achieve high ISR pass rate. The effect of systematic bias (e.g., instability, interconversion) on ISR pass rate is strongly dependent on sample size at intermediate IS imprecision. The results provide an understanding of the effect of IS imprecision on ISR pass rates and a framework for selection of ISR sample sizes.

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REFERENCES

  1. Brockman AH, Hatsis P, Paton M, Wu J-T. Impact of differential recovery in bioanalysis: the example of bortezomib in whole blood. AAPS J. 2007;79:1599–603.

    CAS  Google Scholar 

  2. Silvestero L, Gheorghe MC, Tarcomnicu I, Savu S, Savu SR, Iordachescu A, et al. Development and validation of an HPLC-MS/MS method to determine clopidogrel in human plasma. Use of incurred samples to test back-conversion. J Chromatogr B. 2010;878:3134–42.

    Article  Google Scholar 

  3. Côté C, Lahaie M, Latour S, Bergeron M, Dicaire C, Savoie N, et al. Impact of methylation of acyl glucuronide metabolites on incurred sample reanalysis evaluation; ramiprilat case study. Bioanalysis. 2011;3(9):951–65.

    Article  PubMed  Google Scholar 

  4. Yadav M, Shrivatsav PS. Incurred sample reanalysis (ISR): a decisive tool in bioanalytical research. Bioanalysis. 2011;3(9):1007–24.

    Article  CAS  PubMed  Google Scholar 

  5. Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, et al. Workshop/conference report—quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. AAPS J. 2007;9(1):E30–42.

    Article  PubMed Central  Google Scholar 

  6. Fast DM, Kelley M, Viswanathan CT, O’Shaughnessy J, King SP, Chaudhary A, et al. 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. 2009;11(2):238–41.

    Article  PubMed Central  PubMed  Google Scholar 

  7. Timmerman P, Luedtke S, van Amsterdam P, Brundy-Kloeppel M, Lausecker B, Fischmann S, et al. Incurred sample reproducibility; views and recommendations by the European Bioanalysis Forum. Bioanalysis. 2009;1(6):1049–56.

    Article  CAS  PubMed  Google Scholar 

  8. Savoie N, Booth BP, Bradley T, Garofolo F, Hughes NC, King SP, et al. The 2nd CVG workshop on recent issues in good laboratory practice bioanalysis. Bioanalysis. 2009;1(1):19–30.

    Article  CAS  PubMed  Google Scholar 

  9. Guideline on bioanalytical method validation. Committee for Medicinal Products for Human Use. European Medicine Agency. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf (2011). Accessed September 2013

  10. Notice to guidance document: conduct and analysis of comparative bioavailability studies. Health Canada. http://www.hc-sc.gc.ca/dhp-mps/alt_formats/pdf/prodpharma/applic-demande/guide-ld/bio/gd_cbs_ebc_ld-eng.pdf (2012). Accessed September 2013.

  11. Draft guidance for industry: bioanalytical method validation. US Department of Health and Human Services, Food and Drug Administration. September 2013 http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf (2013). Accessed November 2013.

  12. Rocci Jr ML, Devanarayan V, Haughey DB, Jardieu P. Confirmatory reanalysis of incurred bioanalytical samples. AAPS J. 2007;9(3):E336–43.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Hoffman D. Statistical considerations for assessment of bioanalytical incurred sample reproducibility. AAPS J. 2009;11(3):570–80.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Thway TM, Eschenberg M, Calamba D, Macaraeg C, Ma M, DeSilva B. Assessment of incurred sample reanalysis for macromolecules to evaluate bioanalytical method robustness: effects from imprecision. AAPS J. 2011;13(2):291–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Guidance for industry: bioanalytical method validation. US Department of Health and Human Services, Food and Drug Administration. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM070107.pdf (2001). Accessed September 2013.

  16. Lytle FE, Julian RK, Tabert AM. Incurred sample reanalysis: enhancing the Bland-Altman approach with tolerance intervals. Bioanalysis. 2009;1(4):705–14.

    Article  CAS  PubMed  Google Scholar 

  17. Fu Y, Li W, Smith HT, Tse FLS. An investigation of incurred human urine sample reanalysis failure. Bioanalysis. 2011;3(9):967–72.

    Article  CAS  PubMed  Google Scholar 

  18. Voicu V, Gheorghe MC, Sora LD, Sârbu C, Medvedovici A. Incurred sample reanalysis: different evaluation approaches on data obtained for spironolactone and its active metabolite canrenone. Bioanalysis. 2011;3(12):1343–56.

    Article  CAS  PubMed  Google Scholar 

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ACKNOWLEDGMENTS

We thank Brian Booth, Ethan Stier, and Robert Lionberger for the critical review of the manuscript.

Disclaimer

The views expressed in this article are those of the authors and do not reflect official policy at the FDA. No official endorsement by the FDA is intended or should be inferred.

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Author notes

  1. Barbara M. Davit

    Present address: Merck Sharp & Dohme Corp, Whitehouse Station, New Jersey, USA

Affiliations

  1. Division of Bioequivalence III, Office of Bioequivalence, Office of Generic Drugs, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, Maryland, 20993, USA

    Sriram Subramaniam & Devvrat Patel

  2. Office of Bioequivalence, Office of Generic Drugs, Food and Drug Administration, Silver Spring, Maryland, USA

    Dale P. Conner

Authors
  1. Sriram Subramaniam
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  2. Devvrat Patel
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  3. Barbara M. Davit
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  4. Dale P. Conner
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Corresponding author

Correspondence to Sriram Subramaniam.

Additional information

This work was performed while Barbara Davit was with the FDA.

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Subramaniam, S., Patel, D., Davit, B.M. et al. Analysis of Imprecision in Incurred Sample Reanalysis for Small Molecules. AAPS J 17, 206–215 (2015). https://doi.org/10.1208/s12248-014-9689-1

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  • DOI: https://doi.org/10.1208/s12248-014-9689-1

KEY WORDS

  • bias
  • imprecision
  • incurred sample reanalysis
  • sample size
  • small molecules