Simultaneous determination of six hydrophilic components in rat plasma after oral administration of Jitai tablet by liquid chromatography–electrospray ionization–tandem mass spectrometry: Application to a pharmacokinetic study

A liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) method was developed and validated for the simultaneous determination of amygdalin (ADL), danshensu (DSS), ferulic acid (FA), hydroxysafflor yellow A (HSYA), salvianolic acid A (SAA) and salvianolic acid B (SAB) in rat plasma. Plasma samples were pretreated by protein precipitation with acetonitrile. LC separation was performed on a Zorbax Eclipse Plus C18 column (3.0 mm × 100 mm I.D, 1.8 μm) with gradient elution using a mobile phase consisting of acetonitrile-0.1% formic acid in water at a flow rate of 0.3 mL/min. ESI-MS spectra was acquired in negative ion multiple reaction monitoring mode. The mass transition ion-pair was followed as m/z 456.0 → 323.1, m/z 197.3 → 178.8, m/z 193.0 → 133.9, m/z 611.1 → 325.2, m/z 493.0 → 295.0, and m/z 717.0 → 519.0 for ADL, DSS, FA, HSYA, SAA and SAB, respectively. All analytes showed good linearity over a wide concentration range (r > 0.99). The lower limit of quantification was 7 ng/mL, 2 ng/mL, 4 ng/mL, 1 ng/mL, 2 ng/mL, and 4 ng/mL for ADL, DSS, FA, HSYA, SAA and SAB, respectively. The mean recovery of the analytes ranged from 86.29% to 93.16%. The intra- and inter-day precisions were in the range of 1.50–9.98% and the accuracies were between 91.17% and 99.46%. The validated method was successfully applied to a pharmacokinetic study of the six hydrophilic components in rat plasma after oral administration of Jitai tablet.

► LC–ESI-MS/MS utilized for PK study of six hydrophilic components in Jitai tablet. ► LC–ESI-MS/MS accomplished high–resolution separation and provided lower LLOQs. ► The PK properties of the six components were characterized as rapid oral absorption. ► The method is simple, fast, specific and sensitive.