Assessment of a maslinic acid derivative and its metabolite in rat blood by liquid chromatography coupled with mass spectrometry

Abstract

A new rapid high-throughput liquid chromatography coupled with tandem mass spectrometry method (LCMS/ MS) was developed for the blood assessment of a maslinic acid derivative (Benzyl (2α, 3β) 2,3-diacetoxyolean-12-en-28-amide) and its metabolite. Separation was performed using a Zorbax SB-C18 column with a mobile phase composition of methanol and 0.2% formic acid in water (78:22, v/v) in isocratic flow, with a flow rate of 0.6 mL/min. Detection was performed by multiple reaction monitoring (MRM) mode, monitoring the transitions 338.3; 526.6; 544.6; 527.5; 545.5 m/z derived from 646.6 m/z for the maslinic acid derivative and 409; 338.3; 203; 205; 526.5 m/z derived from 562.6 m/z for its metabolite, using atmospheric pressure chemical positive ionisation source. Single-step protein precipitation with methanol was used for preparation of rat blood samples. The method was validated with respect to selectivity, linearity (r > 0.9937), within-run and between-run precision (CV < 12.0%) and accuracy (bias < 7.9%) over the ranges of 20.5 - 656 ng/mL blood for maslinic acid derivative, and 20 - 640 ng/mL blood for maslinic acid derivative metabolite. The analytical method is simple, fast, selective and is suitable for bioanalysis of maslinic acid derivative and its metabolite from rat blood.