Jiang Tong, Zhu Tong, Teng Fei, Yang Dan, Zhu Jing-Jing, Wang Zhi-Min, Liu Zhi-Gao, Liu Ji-Yan
National Engineering Laboratory of Quality Control Technology of Chinese Materia Medica, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Jiangxi Qiyunshan Food Co., Ltd. Ganzhou 341000, China.
Zhongguo Zhong Yao Za Zhi. 2021 Jun;46(12):2923-2930. doi: 10.19540/j.cnki.cjcmm.20210225.303.
The present study determined the quantitative markers of total proanthocyanidins in the purification of the industrial waste Choerospondias axillaris pericarp based on the comparison results of high-performance liquid chromatography(HPLC) and mass spectrometry(MS) and optimized the purification process with two stable procyanidins as markers. The adsorption and desorption of five different macroporous adsorption resins, the static adsorption kinetics curve of NKA-Ⅱ resin, the maximum sample load, and the gradient elution were investigated. The UPLC-Q-TOF-MS/MS was employed for qualitative analysis of the newly-prepared total proanthocyanidins of C. axillaris pericarp. As revealed by the results, NKA-Ⅱ resin displayed strong adsorption and desorption toward total proanthocyanidins. The sample solution(50 mg·mL~(-1)) was prepared from 70% ethanol crude extract of C. axillaris pericarp dissolved in water and 7-fold BV of the sample solution was loaded, followed by static adsorption for 12 h. After 8-fold BV of distilled water and 6-fold BV of 10% ethanol were employed to remove impurities, the solution was eluted with 8-fold BV of 50% ethanol, concentrated, and dried under reduced pressure, and purified total proanthocyanidin powder was therefore obtained. Measured by vanillin-hydrochloric acid method, the purity and transfer rate of total proanthocyanidins were 47.67% and 59.92%, respectively, indicating the feasibi-lity of the optimized process. UPLC-Q-TOF-MS/MS qualitative analysis identified 16 procyanidins in C. axillaris total proanthocyanidins. The optimized purification process is simple in operation and accurate in component identification, and it can be applied to the process investigation of a class of components that are difficult to be separated and purified. It can also provide technical support and research ideas for the comprehensive utilization of industrial waste.
本研究基于高效液相色谱（HPLC）和质谱（MS）的比较结果，确定了工业废弃物南酸枣果皮纯化过程中原花青素总量的定量标记物，并以两种稳定的原花青素为标记物优化了纯化工艺。研究了5种不同大孔吸附树脂的吸附和解吸性能、NKA-Ⅱ树脂的静态吸附动力学曲线、最大上样量及梯度洗脱。采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）对新制备的南酸枣果皮原花青素总量进行定性分析。结果表明，NKA-Ⅱ树脂对原花青素总量表现出较强的吸附和解吸能力。将南酸枣果皮70%乙醇粗提物用水溶解配制成样品溶液（50 mg·mL⁻¹），上样7倍床体积，静态吸附12 h。先用8倍床体积蒸馏水和6倍床体积10%乙醇去除杂质，再用8倍床体积50%乙醇洗脱，浓缩并减压干燥，得到纯化的原花青素粉末。采用香草醛-盐酸法测定，原花青素总量的纯度和转移率分别为47.67%和59.92%，表明优化工艺可行。UPLC-Q-TOF-MS/MS定性分析鉴定出南酸枣原花青素中有16种原花青素。优化后的纯化工艺操作简单、成分鉴定准确，可应用于一类难分离纯化成分的工艺研究。也可为工业废弃物的综合利用提供技术支持和研究思路。
