黄精粗多糖提取及蒽酮-硫酸分光光度法含量测定

Extraction Process of Crude Polysaccharide from Manyflower Solomonseal Rhizome and Content Determination of Polysaccharide in the Crude Polysaccharide by the Method of Anthrone-sulfuric Acid Spectrophotometric

目的：优选黄精粗多糖的提取纯化工艺并建立蒽酮-硫酸分光光度含量测定法。方法：以黄精提取物中多糖得率为提取工艺评价指标,采用正交实验方法优选黄精的水煎煮提取工艺;采用多级醇沉-Sevage法脱蛋白-活性炭脱色的组合工艺纯化黄精提取物以获得黄精粗多糖;采用蒽酮-硫酸分光光度法测定黄精粗多糖含量。结果：黄精提取物的最佳水煎煮提取工艺为：以8倍量水煎煮,提取3次,每次1小时;黄精提取物经组合工艺纯化得黄精粗多糖,黄精粗多糖相对于黄精提取物、黄精药材计算其得率分别为42.30%和13.35%,粗多糖含量以无水葡糖糖计为98.60%;葡萄糖浓度在5.29~21.16μg·ml-1范围内线性关系良好（r=0.9995,n=7）,平均加样回收率为99.42%,RSD为1.99%（n=9）。结论：本实验优选的黄精粗多糖的提取、纯化工艺稳定、可行、合理;建立的蒽酮-硫酸分光光度法测定黄精粗多糖的方法准确度高、重复性好、操作简单。

Objective： The objective was to optimize the extraction process for crude polysacchafide from Manyflower So- lomonseal Rhizome, to screen the purification process to obtain the crude polysaccharide, and to establish the method of anthrone -sulfuric acid spectrophotometric for the content determination of polysaccharide in the crude polysaccharide. Methods ： The yield of polysaccharide in Manyflower Solomonseal Rhizome extract was used as the index and the best water-decoction technique was optimized by the orthogonal design; the crude polysaccharide was obtained through a combined process which included three steps： the first one was that Manyflower Solomonseal Rhizome extract was precipitated by alcohol repeatedly; the second was that the proteins in the extract was removed by sevage reagen; the third was that the extract was decolorized by activated carbon; the method of anthrone-sulfuric acid spectrophotometric was used to determine the content of polysaccharide in crude polysaccharide. Results ： The opimized parameter resulted when the water consumption was 8times the weight of Manyflower Solomonseal Rhizome, which was decocted three times, once per hour; the crude polysaccharide was obtained from Manyflower Solomonseal Rhizome ex-tract by the combined process. The yield of crude polysaccharide was 42. 30% when it was calculated on the weight of Manyflower Solomonseal Rhizome extract and the yield of crude polysaccharide was 13.35% when it was calculated on the weight of Many- flower Solomonseal Rhizome. The content of crude polysaccharide was 98. 60% which was calculated on the weight of anhydrous glucose. A linearity of glucose was obtained from the range of 5.29 - 21. 16μg ·ml^-1 with a good correlation coefficient （ r= 0. 9995, n=7）. The average recovery was 99.42%, RSD = 1.99% （ n = 9）. Conclusion： The extraction technique for crude polysaccharide from Manyflower Solomonseal Rhizome and the purification technique for crude polysaccharide were stable, feasible and reasonable. The method of anthrone-sulfuric acid spectrophotometric for content determination of polysaccharide in crude pol- ysaccharide was high accuracy, good repetitiveness and simple operation.