Holistic analysis of seven constituents from five medicinal herbs composing Wuhu powders in a single run by ultra performance liquid chromatography: application to quality control study

Abstract

The newly proposed strategy, namely, holistic analysis of multiple constituents from every constituent herb in a single run, was applied for the quality control of traditional Chinese medicine preparation Wuhu powder. Seven bioactive components, i.e., ferulic acid, hydroxysafflor yellow A, prim-O-glucosylcimifugin, 4′-O-β-D-glucosyl-5-O-methylvisamminol, imperatorin, isoimperatorin, and linoleic acid, from 5 medicinal herbs composing Wuhu powder were selected as evaluation markers, and then their quantitative information was obtained in a single run by an UPLC method coupled with photodiode array detection. Optimum separation was achieved on a C₁₈ column (50 × 2.1 mm i.d., 1.7 μm particle) through a gradient delivery of a mixture of acetonitrile and 0.1% aqueous phosphoric acid at a flow rate of 0.3 mL min⁻¹ at 30 °C. The corresponding maximum absorption wavelengths were selected as the detection wavelengths for each analyte. For all of the analytes, a good linear relationship (r of >0.999) was obtained between peak area and concentration. The method was validated for repeatability, precision, stability, accuracy, and selectivity. The RSD values of retention time and peak area were 0–0.6% and 0.7–1.0% for intra-assay, and 0.1–1.0% and 1.6–1.8% for inter-assay precision. Analysis repeatability and stability (24 h) were all lower than 5.0%. The recovery of the method was 96.1–97.7% with a RSD of 2.4–4.0%. The validated method was applied to quality control of Wuhu powder from different production batches.