A validated high-performance liquid chromatographic method with diode-array detection for the estimation of xyloketal B in rat plasma

A sensitive and specific HPLC–UV method was developed and validated for the determination of xyloketal B in rat plasma. Following liquid–liquid extraction, the separation was performed using an isocratic mobile phase of methanol–acetonitrile–water (30/30/40, v/v/v) on a Phenomenex C18 column (4.6mm×250mm, 5μm). The eluent was monitored at 220nm and at a flow rate of 0.8mlmin−1. A linear curve over the concentration range of 1–128μg/ml (r>0.999) was established. The LLOQ of the method was 1μg/ml. Good precision and accuracy at concentrations of 2.5, 25 and 100μg/ml were obtained. The recovery of xyloketal B in plasma was >87.91%. The validated method was found to be specific, precise and accurate in the study. The analytic method was satisfactorily applied to perform preclinical pharmacokinetic study of xyloketal B in rat plasma.