Simultaneous LC–MS–MS Analysis of Danshensu, Salvianolic Acid B, and Hydroxysafflor Yellow A in Beagle Dog Plasma, and Application of the Method to a Pharmacokinetic Study of Danhong Lyophilized Powder for Injection

Abstract

A reliable and sensitive liquid chromatographic–tandem mass spectrometric method, with rutin as internal standard, has been developed and validated for simultaneous determination of danshensu, salvianolic acid B (SAB), and hydroxysafflor yellow A (HSYA) in beagle dog plasma. Plasma samples spiked with the analytes were extracted by solid-phase extraction and the analytes were separated on a 250 × 4.6 mm i.d., 5-μm particle, C₁₈ column with methanol–acetonitrile–0.5% formic acid 20:25:55 (v/v) as mobile phase at a flow rate of 1 mL min⁻¹. LC–MS–MS analysis was performed with a Finnigan TSQ triple-quadrupole tandem mass spectrometer operated in negative-ion selected-reaction-monitoring mode, using electrospray ionization. The accuracy and precision of the method were acceptable and linearity was good over the range 20–4,000 ng mL⁻¹ for danshensu, 50–10,000 ng mL⁻¹ for SAB, and 10–2,000 ng mL⁻¹ for HSYA. The method was successfully applied to a pharmacokinetic study of a traditional Chinese medicinal preparation, Danhong lyophilized powder for injection.