Abstract

To establish an high-performance liquid chromatography-diode array detection (HPLC-DAD) method for the first time to simultaneously determine five major bioactive depsides, salianic acid A , protocatechuic acid, protocatechuic aldehyde, salvianolic acid G and salvianolic acid D in freeze-dried Dan-Shen injection. Chromatographic separation was carried out on a Phenomenex Hydro-C18 reserved-phase column (4.6×250 mm, 4 mm) by stepwise gradient elution with acetonitrile and water (0.1% trifluoroacetic acid, v/v) as the mobile phase. The UV detection wavelength was set at 281 nm for salianic acid A and protocatechuic aldehyde, 260 nm for protocatechuic acid, 228 nm for salvianolic acid G and 322 nm for salvianolic acid D, respectively. Good linear relationships were observed between peak areas and concentrations with r² values above 0.9997 for all the analytes. Average recoveries for the five major actives ranged from 98.3 to 99.4%. This method was rapid, sensitive, accurate, and could be readily applied to the quality assessment of freeze-dried Dan-Shen injection.

Key words: Column liquid chromatography, depsides, freeze-dried Dan-Shen injection, quality control.