Determination of Danshensu, a major active compound of Radix Salvia miltiorrhiza in dog plasma by HPLC with fluorescence detection

A simple and sensitive high performance liquid chromatography method has been developed for the determination of Danshensu (3, 4-dihydroxyphenyllactic acid) in dog plasma. Plasma samples were extracted with ethyl acetate. Analysis of the extracts was performed on a reversed-phase column with an aqueous phosphate buffer-acetonitrile (93:7, v/v) mobile phase, and 4-hydroxybenzoic acid was used as the internal standard. Fluorescence detection at 285 nm (excitation) and 320 nm (emission) was employed. Standard curves were linear in the range from 0.125 to 11.3 microg/mL (regression coefficient r > 0.993) on three different days. Mean recovery was determined as 96.4% by analysis of plasma standard containing 0.63, 5.65 and 11.3 microg/mL of Danshensu. The inter-day precision (RSD) ranged from 3.4 to 8.6% at concentrations of 0.125, 1.88, 6.28 and 11.3 microg/mL, and the intra-day precision was better than 7.2%. The detection and quantitation limits were 0.063 and 0.125 microg/mL, respectively. This validated assay was applied to the determination of Danshensu concentration in dog plasma after oral administration of Radix Salvia miltiorrhiza extracts.