赶黄草总黄酮树脂精制工艺与检测方法.PDFVIP

基 因组学与应用生物学，2019 年，第 38 卷，第 5 期，第 2230-2237 页 研究报告 Research Report 赶黄草总黄酮树脂精制工艺与检测方法 屈晓宇 高秋玉 邓小宽 高平 * 四川大学生命科学学院, 生物资源与生态环境教育部重点实验室, 成都, 610065 * 通信作者, gaop@scu.edu.cn 摘 要 本研究以总黄酮吸附量和解析率为检测指标，结合动态洗脱考察结果，考察 11 种树脂对赶黄草总 黄酮的富集精制能力。优化树脂类型，选定了对总黄酮进行富集纯化较好的树脂 HPD450 ；采用单因素及正交 实验，确定最佳工艺条件：上样量 10 mL，以 6 mL/min 的流速 8 BV 水、6 BV 80% 甲醇溶液洗脱，pH 值为 6~7。 以此方法得到 90% 以上 的赶黄草总黄酮，实验结果 良好，总黄酮精制工艺成效显著。用 HPLC 法精确测定赶 黄草中槲皮苷、PGHG 和 ThA 三个指标性黄酮成分，最佳色谱条件为：C18 色谱柱，检测波长 280 nm ，进样量 5 μL，流动相:乙腈 -0.05%磷酸溶液(0~25 min: 12%~45% 乙腈; 25~40 min: 45%~70%乙腈)梯度洗脱 ；精确测 定赶黄草全草 中 3 种黄酮成分的含量。采用分光光度法检测，指导优化总黄酮精制工艺，并 以 HPLC 法精确 测定三种黄酮代表成分。比较两种方法测定结果相一致，互为补充 ；故在工业生产时，可 以分光光度法指导生 产，以 HPLC 法精确定量测定。 关键词 赶黄草, 总黄酮, 分光光度法, HPLC Refining Technology and Detection Method of Total Flavonoids Resin from Penthorum chinense Pursh Qu Xiaoyu Gao Qiuyu Deng Xiaokuan Gao Ping * Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065 * Corresponding author, gaop@scu.edu.cn DOI: 10.13417/j.gab.038.002230 Abstract In this study, the adsorption quantity and resolution of total flavonoids were used as indicators, and combined with the results of dynamic elution, the enrichment and refining capacity of 11 resins for total flavonoids of Penthorum chinense Pursh was investigated. To optimize the type of resin, HPD450 resin was selected as a better resin for enrichment and purification of total flavonoids. The optimum technological conditions were determined by single factor and orthogonal experiments: 10 mL sample size, 8 BV water at 6 mL/min flow rate, 6 BV 80% methanol solution elution, and pH 6~7. More than 90% of total flavonoids were obtained by this method. The experimental results were good and the refining process of total flavonoids was remarkable. Quercetin, PGHG and ThA were accurately determined by HPLC. The optimum parameter