College of Pharmaceutical Sciences, Key Laboratory of Pharmaceutical Quality Control of Hebei Province, Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, Key Laboratory of Public Health Safety of Hebei Province, State Key Laboratory of New Pharmaceutical Preparations and Excipients, Hebei University, Baoding 071002, China.
College of Pharmaceutical Sciences, Key Laboratory of Pharmaceutical Quality Control of Hebei Province, Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, Key Laboratory of Public Health Safety of Hebei Province, State Key Laboratory of New Pharmaceutical Preparations and Excipients, Hebei University, Baoding 071002, China.
J Pharm Biomed Anal. 2024 Mar 15;240:115947. doi: 10.1016/j.jpba.2023.115947. Epub 2023 Dec 29.
Huangqi Liuyi Decoction, a famous classical Chinese prescription, shows significant curative effect on diabetes and its complications, in which calycosin-7-glucoside, liquiritin and glycyrrhizic acid are the main components that playing these mentioned pharmacological activity, under the synergistic action of various other ingredients in the decoction. However, there are significant differences in the content of active compounds in Chinese medicinal materials, which mainly due to origin, picking seasons, and processing methods. Hence, the accurate content of the glycosides is the prerequisite for ensuring the pharmacological efficacy. Aiming at establishing an efficient extraction and determination method for accurate quantitative analysis of calycosin-7-glucoside, liquiritin and glycyrrhizic acid in Huangqi Liuyi Decoction, an on line solid-phase extraction-high-performance liquid chromatography method was developed, using a homemade bio-based monolithic adsorbent. The bio-based adsorbent was prepared in a stainless steel tube, using bio-monomers of methyleugenol and S-allyl-L-cysteine, which effectively reduced the dependence of the polymer field on non-renewable fossil resources and reduced carbon emissions. Furthermore, the prepared adsorbent owned abundant chemical groups, which can produce interactions of hydrogen bond, dipole-dipole, π-π and hydrophobic force with the target glycosides, thus improving the specific recognition ability of the adsorbent. The experiments were carried out on an LC-3000 HPLC instrument with a six-way valve. Methodology validation indicates that the recovery is in the range of 97.0%-103.4% with the RSD in the range of 1.6%-4.0%, due to the specific selectivity of the bio-based monolithic adsorbent for these three glycosides, and good matrix-removal ability for Huangqi Liuyi decoction. The limit of detection is 0.17, 0.50 and 0.33 μg/mL for calycosin-7-glucoside, liquiritin and glycyrrhizic acid, respectively, and the limit of quantitation is 0.50, 1.50 and 1.00 μg/mL, respectively, with the linear range of 2-200 μg/mL for calycosin-7-glucoside, and 5-500 μg/mL for liquiritin and glycyrrhizic acid. The present work provided a simple and efficient method for the extraction and determination of glycosides in complex medicinal plants.
黄芪六君子汤是一种著名的中药方剂，对糖尿病及其并发症具有显著的疗效，其中芒柄花苷、甘草苷和甘草酸是发挥这些药理活性的主要成分，这些成分在方剂中的各种其他成分的协同作用下发挥作用。然而，中药材中活性化合物的含量存在显著差异，主要是由于来源、采摘季节和加工方法的不同。因此，准确的糖苷含量是保证药理功效的前提。本研究旨在建立一种高效的提取和测定方法，用于准确定量分析黄芪六君子汤中芒柄花苷、甘草苷和甘草酸的含量，开发了一种在线固相萃取-高效液相色谱法，使用自制的生物基整体式吸附剂。生物基吸附剂是在不锈钢管中制备的，使用的生物单体为丁香酚和 S-烯丙基-L-半胱氨酸，这有效地降低了聚合物领域对不可再生化石资源的依赖，并减少了碳排放。此外，所制备的吸附剂具有丰富的化学基团，可与目标糖苷产生氢键、偶极-偶极、π-π和疏水相互作用，从而提高吸附剂的特异性识别能力。实验在 LC-3000 HPLC 仪器上进行，采用六通阀。方法学验证表明，回收率在 97.0%-103.4%范围内，相对标准偏差在 1.6%-4.0%范围内，这是由于生物基整体式吸附剂对这三种糖苷具有特异性选择性，并且对黄芪六君子汤具有良好的基质去除能力。芒柄花苷、甘草苷和甘草酸的检测限分别为 0.17、0.50 和 0.33μg/mL，定量限分别为 0.50、1.50 和 1.00μg/mL，芒柄花苷的线性范围为 2-200μg/mL，甘草苷和甘草酸的线性范围为 5-500μg/mL。本工作为复杂药用植物中糖苷的提取和测定提供了一种简单高效的方法。
