HPLC simultaneous determination of five constituents in Salvia miltiorrhiza phenolic acids extract by wavelength switching

AIM To establish an HPLC method for simultaneously determining five constituents(sodium danshensu,protocatechuic aldehyde,rosmarinic acid,lithospermic acid,salvianolic acid B) in Salvia miltiorrhiza phenolic acids extract.METHODS The separation was performed on a Phenomsil-C18 ODS(250 mm×4.6 mm,5 μm) column with the gradient elution of acetonitrile-water of 0.5% phosphoric acid at the flow rate of 1.0 mL/min.The column temperature was set at 30 ℃.Detection wavelength was 280 nm in 0-50 min,330 nm in 50-68 min,286 nm in 68-90 min.RESULTS Sodium danshensu,protocatechuic aldehyde,rosmarinic acid,lithospermic acid and salvianolic acid B had good linearity in the ranges of 0.033 96-0.339 6 μg,0.016 60-0.166 0 μg,0.092 4-0.924 μg,0.084 7-0.847 μg,1.300-13.00 μg,repectively.The average recoveries of constituents were 99.2%-100.2%,with RSDs less than 2.3%.The contents of five constituents in Salvia miltiorrhiza phenolic acids extract from seven sources showed differences.CONCLUSION The developed method is rapid,accurate with high roproducibility and stability,and helpful to control the quality of Salvia miltiorrhiza phenolic acids extract.