Objective:To establish an HPLC method to determine simultaneously the content of four water-soluble chemical constituents (salvianolic acid D

rosmarinic acid

lithospermic acid and salvianolic acid B) in salvianolic acid extracts

and to compare their content discrepancy of salvianolic acid extracts from 6 batches. Method: Chromatographic column was Venusil XBP C18 (4.6 mm×250 mm

5 μm) at the gradient elution mode with the flow rate of 1.0 mL·min-1

mobile phase was phosphoric acid-water (0.02∶100) and phosphoric acid-acetonitrile-water (0.02∶80∶20). Column temperature was kept at 30 ℃ and the detector was set at 280 nm. Result: The regress equation for salvianolic acid D was A=13 979C-1 739.2

R2=0.999 6

and the linear range was 2.11-21.10 mg·L-1. Rosmarinic acid was A=42 007C-448.58

R2=0.999 7

and the linear range was 3.94-39.40 mg·L-1. Lithospermic acid was A=23 061C-6 319.2

R2=0.999 6

and the linear range was 4.14-41.40 mg·L-1. Salvianolic acid B was A=22 186C-21 435

R2=0.999 8

and the linear range was 53.90-539.00 mg·L-1. The average recoveries of salvianolic acid D

rosmarinic acid

lithospermic acid and salvianolic acid B were 100.12%

100.60%

102.47% and 104.53%

RSD 1.52%

1.83%

1.53% and 2.00%

respectively. Conclusion: The method is simple and accurate for determination of salvianolic acid D

rosmarinic acid

lithospermic acid and salvianolic acid B in the salvianolic acid extract.