Holistic analysis of Liuwei Dihuang pills using ultrasonic cell grinder extraction and ultra-performance liquid chromatography

Abstract

An efficient holistic analysis strategy was developed and validated for quality control of the traditional Chinese medicine (TCM) preparation, Liuwei Dihuang pills (LWDHPs). In this strategy, eight bioactive components, i. e. allantoin, morroniside, loganin, peoniflorin, acteoside, paeonol, alisol B 23-acetate and pachymic acid from the six medicinal herbs composing LWDHPs were selected as the evaluation markers, and then they were simultaneously extracted by an ultrasonic cell grinder extraction (UCGE) method and separated in a single run by an ultra-performance liquid chromatographic method coupled with photodiode array detection (UPLC-PDA). Response surface methodology (RSM), a multivariate experimental design technique, was used to optimize the UCGE parameters based on Box-Behnken design (BBD) model. The optimum extraction conditions were obtained at a ratio of solvent to solid 45 mL/g, extraction time of 40 min and extraction temperature of 20℃. Under such conditions, the higher yields of the eight analytes were acquired compared with the traditional extraction methods, indicating that UCGE was a fast, easy and efficient method for extracting bioactive constituents from LWDHPs. Chromatographic separation was achieved on an HSS T3 column (50 mm×2.1 mm, 1.8 μm) using a linear gradient elution of acetonitrile and water. Because of the different UV characteristics of these components, five detection wavelengths were used for quantitative analysis. The proposed method was validated through linearity, limits of detection, limits of quantification, precision, stability, repeatability, and accuracy. The validated method was applied to analyze LWDHPs, which provided a reference for the quality evaluation of LWDHPs.