A Simple and Reliable Method for Distinguishing Danshen in Salvia: Simultaneous Quantification of Six Active Compositions by HPLC

A simple and reliable method for distinguishing Danshen is important to evaluate the quality and clinical efficiency of these species. An HPLC method was developed for the determination of protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone, tanshinone I and tanshinone IIA in 23 samples of Salvia. The analytes were separated on an Agilent XDB C18 reversed-phase column coupled with a Phenomenex C18 guard column using a gradient elution of acetonitile-0.1% aqueous phosphoric acid as the mobile phase at a flow rate 0.8 mL/min and UV detection at 280 nm. The method allowing the simultaneous quantification of six major active compositions was optimized and validated for linearity, precision, accuracy and limits of detection (LOD) and quantification. The LOD ranged from 0.019 to 0.850 µg/mL (R(2) ≥ 0.9998). Accuracy, precision and reproducibility were all within the required limits. The average recovery between 96.49 and 102.16% and the relative standard deviations were <3.01%. Based on the six compositions content and clustering result, this research results suggest that these six major active compositions could be distinguishing markers for Danshen and non-Danshen.