赶黄草总黄酮树脂精制工艺与检测方法

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5μL,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。

Refining Technology and Detection Method of Total Flavonoids Resin from Penthorum chinense Pursh

In this stu dy, 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 m L/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 parameters of chromatographic conditions were as follows: C18 chromatographic column, detection wavelength 280 nm, injection volume 5 L,mobile phase: acetonitrile-0.05% phosphoric acid solution(0~25 min: 12%~45% acetonitrile, 25~40 min: 45%~70%acetonitrile) gradient elution. accurate determination of three flavonoids in Herba Phellodendron. The contents of three flavonoids in Penthorum chinense Pursh were determined accurately. Spectrophotometric determination was used to guide the optimization of total flavonoids refining process;and the representative components of three flavonoids were accurately determined by HPLC. The results of the two methods are consistent and complementary. Therefore, in industrial production, Spectrophotometry could guide the production, and accuratequantitative determination by HPLC.