School of Pharmacy, Shenyang Pharmaceutical University, No.26 Huatuo Rd, High & New Tech Development Zone, 117004, Benxi, Liaoning Province, PR China.
School of Chinese Medicine, Shenyang Pharmaceutical University, No.26 Huatuo Rd, High & New Tech Development Zone, 117004, Benxi, Liaoning Province, PR China.
J Pharm Biomed Anal. 2020 Jun 5;185:113228. doi: 10.1016/j.jpba.2020.113228. Epub 2020 Mar 4.
Deep eutectic solvent (DES) combined with ultrasound-assisted extraction (UAE) was successfully developed and fully validated to simultaneously determine Icarrin, IcarisidⅡ, Epimcdin A, Epimcdin B and Epimcdin C for the quality evaluation of Herba Epimedii. Twelve kinds of DESs were initially screened, and then the effective extraction was achieved by the tailor-made DES consisting of the mixture of l-proline and ethylene glycol with the molar ratio of 1:4 in this study. The optimal conditions were further optimized by the orthogonal experimental design (OED). 0.2 g sample powder was ultrasonic extracted by using 4.00 mL of aqueous solution containing 70 % (v/v) the above DES for 45 min, resulting to the optimum extraction efficiency. The FT-IR and NMR spectra showed the chemical structural characteristic correlation between l-proline and ethylene glycol, and could infer the formation of hydrogen bonds between the hydroxyl group of ethylene glycol and the nitrogen atom of l-proline. The hierarchical cluster analysis (HCA) was further processed for the quality evaluation of Herba Epimedii. Finally, DES could be used to distinguish different origins and different kinds of Herba Epimedii, and to evaluate the quality of Herba Epimedii. This method provided good linearity, precision and accuracy. The recoveries of the five main bioactive flavonoids in Herba Epimedii were within the range of 88.5-107.7 % (RSD less than 3.4 %). Compared to the traditional extraction method of Icarin in the Chinese Pharmacopoeia (2015 edition), the solvent consumption was decreased by 80 % and the extraction time was shortened by 25 %, leading to more efficient and more convenient of this DES-UAE method. This work indicated that DES would be a promising high effective solvent for the analytical sample preparations of plant herbs, and it might have a broad application in the quality control of traditional Chinese medicines.
深共晶溶剂（DES）结合超声辅助提取（UAE）被成功开发并全面验证，可同时测定淫羊藿中的朝藿定 C、朝藿定 B、朝藿定 A、淫羊藿苷 I 和淫羊藿次苷Ⅱ，用于淫羊藿的质量评价。本研究初步筛选了 12 种 DES，然后通过定制的 DES 实现有效提取，该 DES 由 l-脯氨酸和乙二醇以 1:4 的摩尔比混合而成。通过正交实验设计（OED）进一步优化了最佳条件。0.2 g 样品粉末用 4.00 mL 含有 70%（v/v）上述 DES 的水溶液超声提取 45 min，达到最佳提取效率。FT-IR 和 NMR 谱表明 l-脯氨酸和乙二醇之间存在化学结构特征相关性，并可以推断乙二醇的羟基和 l-脯氨酸的氮原子之间形成氢键。进一步进行层次聚类分析（HCA），用于淫羊藿的质量评价。最后，DES 可用于区分不同来源和不同种类的淫羊藿，并评价淫羊藿的质量。该方法具有良好的线性、精密度和准确度。淫羊藿中 5 种主要生物活性黄酮的回收率在 88.5-107.7%（RSD 小于 3.4%）范围内。与中国药典（2015 年版）中 Icarin 的传统提取方法相比，溶剂消耗减少了 80%，提取时间缩短了 25%，使得这种 DES-UAE 方法更加高效和方便。本工作表明，DES 将成为植物草药分析样品制备的一种有前途的高效溶剂，可能在中药质量控制中具有广泛的应用。
