Validation and Application of an Rapid HPLC–MS Method for the Determination of Salvianic Acid A in Human Plasma

A rapid liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI–MS-MS) method was developed and validated for the determination of salvianic acid A in plasma of Chinese healthy subjects after oral administration of Qishenyiqi dropping pills. After liquid–liquid extraction with ethyl acetate, salvianic acid A was chromatographed on a Agilent Zorbax XDB-C18 column using a gradient mobile phase consisting of water (0.1% formic acid) – acetonitrile (0.1% formic acid) at a flow rate of 0.45 mL/min. The detection was performed in multiple reaction monitoring mode, using the transitions of m/ z1 96.9!134.8 and m/ z3 20.9!151.9 for salvianic acid A and chloroamphenicol, respectively. The method was linear over the range of 0.50–500 ng/mL using only 100 mL of plasma and the lower limit of quantification was 0.50 ng/mL. Intra-day and inter-day precisions (in terms of % RSD) were all <15% and the accuracies (in terms of % RE) were within the range of+15%, and recoveries were between 85.0 and 115%. The validated method was successfully applied to pharmacokinetic study of Qishenyiqi dropping pills in Chinese healthy subjects. After oral administration, Tmax and Cmax values were 1.33+ 0.52 h and 21.1+ 3.92 ng/mL, respectively. Plasma concentrations declined with t1/2Z of 1.76+ + 0.33 h.