Application of in-sample calibration curve methodology for regulated bioanalysis: Critical considerations in method...

Area of Science:

• Analytical Chemistry
• Bioanalysis
• Mass Spectrometry

Background:

• Traditional LC-MS/MS bioanalysis relies on external calibration curves for accurate analyte quantification.
• A novel in-sample calibration curves (ISCC) methodology offers a potential alternative, utilizing naturally occurring isotopologues for quantification.
• This ISCC approach has broad applications in biomarker measurement, quantitative proteomics, and clinical diagnostics.

Purpose of the Study:

• To assess the feasibility of the ISCC-LC-MS/MS methodology in regulated bioanalysis.
• To validate the ISCC-LC-MS/MS approach using a drug candidate, BMS-984478, in human plasma.
• To propose method validation procedures for the ISCC strategy for industry consideration.

Main Methods:

• Development and validation of an LC-MS/MS method employing the ISCC strategy for BMS-984478 quantification.
• Quantitative analysis of BMS-984478 in human plasma over a concentration range of 1.33-993.42 ng/mL.
• Comparative performance assessment against a previously validated external calibration curve method.

Main Results:

• A robust ISCC-LC-MS/MS method was successfully developed and validated for BMS-984478 in human plasma.
• The validated ISCC-LC-MS/MS method exhibited performance equivalent to the conventional external calibration curve method.
• Critical considerations for method development, validation, and sample analysis using ISCC were discussed.

Conclusions:

• The ISCC-LC-MS/MS methodology is a viable and promising approach for regulated bioanalysis.
• This novel approach offers unique advantages and significant potential for widespread application in quantitative analysis.
• The ISCC-LC-MS/MS methodology may revolutionize quantitative bioanalytical strategies.