Aim/Background:

To simultaneously measure bempedoic acid and its primary metabolite, bempedoic acid metabolite-ESP15228, in human plasma via UPLC-MS/MS, the QuEChERS extraction tactic is not suitable. Instead, employ a tailored bioanalytical approach.

Materials and Methods:

The QuEChERS technique was employed to extract human plasma, followed by analysis using LC-MS/MS. Separation of compounds was attained using a Zorbax C18 analytical column (50 mm×2.1 mm, 1.7 μm), with retention times of 4.35 and 5.58 min, respectively. The mobile phase consisted of acetonitrile (80%) and 10 mm ammonium acetate (20%), flowing at a rate of 0.80 mL/min. Ionization of the analyte occurred in the TQD’s electrospray ionization ion source, with detection of its active metabolite achieved using Multiple Reaction Monitoring (MRM) in adverse mode.

Results:

Linear calibration curves for bempedoic acid and its metabolite, ESP15228, in human plasma were established over ranges of 201.639-36115.241 ng/mL and 30.027-4039.748 ng/mL, respectively, with correlation coefficients (r²) exceeding 0.9970, indicating uniform and repeatable linearity. Interassay Coefficients of Variation (CV) for bempedoic acid ranged from 0.1% to 1.2%, while those for ESP15228 ranged from 0.7% to 4.6%. Intra-assay CVs for bempedoic acid and ESP15228 were 0.6% to 0.2% and 1.2% to 6.7%, respectively. Recovery rates for bempedoic acid and ESP15228 were 67.69% and 66.33%, respectively, demonstrating efficient extraction.

Conclusion:

The study demonstrates that the employed approach is sensitive, reliable and validated to fulfill legal requirements, rendering it suitable for Bioavailability-Bioequivalence (BA-BE) research. This underscores its potential as a robust method for assessing the pharmacokinetics and bioequivalence of bempedoic acid and its metabolite, ESP15228, facilitating efficient and accurate evaluation of their therapeutic effects and formulation comparisons.