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Bioanalytical method validation design for the simultaneous quantitation of analytes that may undergo interconversion
1Bioanalytical Research, Metabolism and Pharmacokinetics, Bristol-Myers Squibb Pharmaceutical Research Institute, New Brunswick, NJ 08903-0191, USA. jemalm@bms.com
Journal of Pharmaceutical and Biomedical Analysis
|May 18, 2000
Summary
Accurate bioanalytical methods require careful selection of calibration standards and quality control (QC) samples for interconverting analytes. Matching the analyte ratio in QC samples to calibration standards ensures method accuracy, even with unavoidable interconversion.
Area of Science:
- Bioanalytical Chemistry
- Pharmacokinetics
- Analytical Method Development
Background:
- Quantitative determination of interconverting analytes in biological samples presents challenges.
- Minimizing analyte interconversion during bioanalytical method steps is crucial but may be unavoidable.
Purpose of the Study:
- To investigate the impact of calibration standard and quality control (QC) sample composition on the accuracy of bioanalytical methods for interconverting analytes.
- To determine optimal strategies for developing accurate methods for simultaneous determination of interconverting analytes, using a delta-hydroxy acid and its delta-lactone as models.
Main Methods:
- Evaluated method performance (accuracy, precision) under varying conditions of analyte interconversion (minimized, enhanced conversion of lactone to acid, enhanced interconversion).
- Assessed the effect of differing analyte concentration ratios between QC samples and calibration standards on method accuracy.
- Utilized model compounds: a delta-hydroxy acid (analyte 1) and its delta-lactone (analyte 2).
Main Results:
- Method performance was unacceptable when the analyte ratio in QC samples differed from calibration standards under conditions facilitating interconversion.
- Acceptable method performance was achieved when the analyte ratio in QC samples matched that of calibration standards, regardless of interconversion conditions.
- Identical interconversion in QC samples and calibration standards compensated for analyte transformation, ensuring accuracy.
Conclusions:
- Judicious selection of calibration standards and QC samples, matching analyte concentration ratios, is essential for accurate bioanalytical determination of interconverting analytes.
- Bioanalytical methods should include QC samples with varying analyte ratios, including single-analyte samples, to accommodate unknown test sample compositions.
- Optimizing conditions to minimize interconversion of the expected major analyte is recommended, alongside robust QC strategies.