Simultaneous determination of twelve bioactive constituents in Buyang Huanwu decoction by HPLC-DAD-ELSD and HPLC-TOF/MS.

Buyang Huanwu Decoction (BYHYD) is a classical traditional Chinese medicinal prescription that has been widely used for treating cerebrovascular illnesses for hundreds of years. In this study, a comprehensive analytical method has been developed for quantitative analysis of the major constituents in BYHWD. This method was based on high-performance liquid chromatography coupled to a diode array and evaporative light scattering detectors (HPLC-DAD-ELSD) on a common reverse-phase C(18) column. HPLC coupled with on-line time-of-flight mass spectrometry (HPLC-TOF/MS) was additionally adopted to provide further validation for the constituents. It was found that 0.3% aqueous formic acid and acetonitrile was the optimum mobile phase for gradient elution. This method, which showed excellent precision and accuracy, was successfully applied to quantify the bioactive constituents in six BYHWD products. The validated HPLC-DAD-ELSD method, together with the HPLC-TOF/MS analysis, provided a new basis for assessing the quality of traditional Chinese medicinal prescription consisting of many bioactive components.