Determination of a novel anticancer AMPK activator hernandezine in rat plasma and tissues with a validated UHPLC–MS/MS method: Application to pharmacokinetics and tissue distribution study

Abstract

Hernandezine, a novel anticancer AMPK activator, is a major active constituent of Thalictrum Ranunculaceae. A simple, specific and sensitive liquid chromatography tandem mass spectrometry (LC–MS/MS) method has been developed and validated for the quantification of hernandezine in rat plasma and tissues after intravenous administration. Sample preparation was carried out through a protein-precipitation extraction with acetonitrile using tetrandrine as internal standard (IS). The chromatographic separation was achieved by using an Agilent ZORBAX Eclipse Plus C18 column with a mobile phase of acetonitrile and water (containing 10mM ammonium acetate) in an isocratic elution way. The mass spectrometry (MS) analysis was conducted in positive ionization mode with multiple reaction monitoring (MRM) transitions at m/z 653.4→411.2 for hernandezine and m/z 623.3→381.3 for tetrandrine (IS). Calibration curves were linear over the ranges of 20.0–4000ng/ml f or both plasma samples and tissue samples (r>0.991). The lower limit of quantification (LLOQ) was 20.0ng/ml. The intra-day and inter-day precision (RSD%) were less than 14.0%, while the accuracy was ranged from 85.2% to 114.9%. Finally, this developed method was successfully applied in the pharmacokinetics and tissue distribution study of hernandezine after intravenous administration.