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.