Abstract
Paeoniae Radix Alba (PRA), an herbal drug produced from the root of Paeonia lactiflora Pall., is widely used in many herbal medicine prescriptions/preparations. Since the pharmacological effects of PRA come from multiple chemical components, it is important to establish a method for the determination of those components in PRA extracts with simple operation and low cost, which is more suitable to evaluate the quality of PRA extracts and optimize the extraction process. This work introduced the quantitative analysis of multicomponents with a single-marker (QAMS) method for the simultaneous determination of eleven bioactive components in PRA extracts, including gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, ethyl gallate, galloylpaeoniflorin, pentagalloylglucose, benzoic acid, benzoylpaeoniflorin, and paeonol. In the QAMS method established based on high performance liquid chromatography with diode array detection, only the reference substance of paeoniflorin was needed, and the other ten components were determined based on their relative correction factors (RCFs) to paeoniflorin. Moreover, the repeatability and robustness of the RCFs were studied with different column temperatures, detection wavelengths, flow rates, column types, and instruments. In method validation, good linearity (r > 0.999), stability, repeatability (RSD < 1.9%), and accuracy (recoveries within 96.1%–105.5%) were shown. Sample analyses showed that the QAMS method was consistent with the conventional external standard method. The established method provided a comprehensive, efficient, and low-cost tool for the routine quality evaluation of PRA extracts.
Highlights
Paeoniae Radix Alba (PRA) has a long history of medicinal use in China, which is produced from the root of Paeonia lactiflora Pall. (RPLP) by removing its root bark, boiling in water, and drying in the sunlight
Chinese Medical University). e reference substance of gallic acid was purchased from Push Bio-Technology Co., Ltd. (Chengdu, Sichuan, China). e reference substances of oxypaeoniflorin, albiflorin, paeoniflorin, and benzoylpaeoniflorin were purchased from Victory Biological Technology Co., Ltd. (Chengdu, Sichuan, China). e reference substance of catechin was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). e reference substance of pentagalloylglucose was purchased from Xunchen Biological Technology Co., Ltd. (Chengdu, Sichuan, China). e reference substances of ethyl gallate and benzoic acid were purchased from Rhawn Science and Technology Co., Ltd. (Shanghai, China). e reference substance of paeonol was purchased from Jianglai BioTechnology Co., Ltd. (Shanghai, China)
Optimization of Analytical Conditions. e effect of acid added to the mobile phase on the separation was compared first, since the retention behavior of phenols and pentagalloylglucose in RP-HPLC columns can be significantly affected by the pH of the mobile phase [1, 21]
Summary
Paeoniae Radix Alba (PRA) has a long history of medicinal use in China, which is produced from the root of Paeonia lactiflora Pall. (RPLP) by removing its root bark, boiling in water, and drying in the sunlight. Since some of the standard substances like oxypaeoniflorin, galloylpaeoniflorin, and benzoylpaeoniflorin are expensive as well as difficult to be prepared, the cost of the analysis will be relatively high, if multiple components are quantified using the conventional external standard method (ESM). Erefore, it is necessary to find a more low-cost and convenient method to evaluate the quality of PRA extracts. For a better quality evaluation of PRA extracts with simple operation and low cost, this work developed an HPLC method to determine as many as eleven components, including gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, ethyl gallate, galloylpaeoniflorin, pentagalloylglucose, benzoic acid, benzoylpaeoniflorin, and paeonol (Figure 1), using only one reference substance of paeoniflorin, and investigated the feasibility and applicability of the QAMS method. Due to its low requirements on instruments and standard references, the proposed method is deemed to be convenient to use in future researches related to PRA and RPLP
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