Jia Li, Wang Liming, Zhang Xiaoxiao, Zhang Qingrui, Lei Peng, Chang Yanxu, Han Lifeng, Chai Xin, Yang Wenzhi, Wang Yuefei, Jiang Miaomiao
National Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Haihe Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
J Pharm Anal. 2024 Nov;14(11):101002. doi: 10.1016/j.jpha.2024.101002. Epub 2024 May 11.
In this study, 34 deep eutectic solvents (DESs) were successfully prepared for the extraction of proanthocyanidin from Rhodiolae Crenulatae Radix et Rhizomes. The extraction process was optimized using single factor exploration and Box-Behnken design-response surface analysis. The extraction rate was significantly improved when the molar ratio of choline chloride to 1,3-propanediol was 1:3.5 and the water content was 30% (/) in DESs. AB-8 macroporous resin and ethyl acetate were used for separation and refining, and the oligomer-rich proanthocyanidin components were eventually obtained. The ultraviolet (UV) and infrared (IR) spectra showed that the proanthocyanidins were mainly composed of catechin and epicatechin. To further clarify the chemical composition of proanthocyanidin, an ion scan list containing 156 proanthocyanidins precursors was obtained by constructing a proanthocyanidins structural library and mass defect filtering (MDF) algorithm, combined with the full mass spectrometry (MS)/dd-MS scan mode that turns on the "if idle pick others" function. By using ultra-high performance liquid chromatography and high-resolution MS (UHPLC/HRMS), the analysis used both targeted and non-targeted methods to detect proanthocyanidins. Finally, 50 oligomeric proanthocyanidin (OPC) compounds were identified, including 7 monomers, 22 dimers, 20 trimers, and 1 tetramer, most of which were procyanidins of proanthocyanidins (84%), and a small amount of prodelphinidin (14%) and other types of proanthocyanidins (2%), which enabled the systematic characterization of proanthocyanidin components from Rhodiolae Crenulatae Radix et Rhizomes. Meanwhile, the comparison with the grape seeds OPCs standard (United States Pharmacopeia) revealed that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes were more abundant, suggesting that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes has promising applications.
在本研究中，成功制备了34种低共熔溶剂（DESs）用于从红景天根茎中提取原花青素。采用单因素探索和Box-Behnken设计-响应面分析对提取工艺进行优化。当氯化胆碱与1,3-丙二醇的摩尔比为1:3.5且DESs中的含水量为30%（/）时，提取率显著提高。使用AB-8大孔树脂和乙酸乙酯进行分离精制，最终得到富含低聚物的原花青素组分。紫外（UV）和红外（IR）光谱表明，原花青素主要由儿茶素和表儿茶素组成。为进一步阐明原花青素的化学组成，通过构建原花青素结构库和质量缺陷过滤（MDF）算法，结合开启“空闲时选其他”功能的全质谱（MS）/dd-MS扫描模式，获得了一个包含156种原花青素前体的离子扫描列表。通过超高效液相色谱和高分辨率质谱（UHPLC/HRMS），该分析采用靶向和非靶向方法检测原花青素。最终鉴定出50种低聚原花青素（OPC）化合物，包括7种单体、22种二聚体、20种三聚体和1种四聚体，其中大部分是原花青素的原花青素（84%），少量是原花青定（14%）和其他类型的原花青素（2%），这使得能够系统地表征红景天根茎中原花青素的组分。同时，与葡萄籽OPCs标准品（美国药典）的比较表明，红景天根茎中的原花青素更为丰富，这表明红景天根茎中的原花青素具有广阔的应用前景。
