A validated UHPLC-MS/MS method for measurement of pharmacokinetics and tissue distribution of trolline in rat

As a novel alkaloid, trolline is a potential methionine aminopeptidase Ⅱ inhibitor. However, up to now, no informations about the quantification of trolline were available in biosamples. In this study, a simple, specific and sensitive analytical method based on UHPLC-MS/MS method has been established and validated for determination of trolline in rat plasma and tissues after intravenous administration. Sample preparation was carried out by a simple liquid-liquid extraction and carbamazepine was used as internal standard (I.S.). Chromatographic separation was achieved by using a Waters BEH C₁₈ column and involving the optimized mobile phase of 0.1% formic acid aqueous solution and acetonitrile with gradient elution flow of 0.20 ml/min. Trolline and I.S. were detected by multiple reaction monitoring (MRM) modes with positive electrospray ionization and transitions at m/z 220.0→136.8 for trolline and m/z 237.0→193.9 for carbamazepine (I.S.). Good linearity was ranged from 10.0 ng/ml to 4000 ng/ml for trolline both in plasma and various tissues. The lower limit of quantification (LLOQ) was 10 ng/ml in all samples. The intra- and inter-day precision (RSD%) were below 11.3% and the accuracy (RE%) was ranged from -10.2% to 12.3%. The validated method was successfully applied to the pharmacokinetics and tissue distribution study of trolline after intravenous administration.