响应面法优化水提醇沉玄参多糖工艺及其含量分析  

Response Surface Methodology for Optimization of Water Extract and Alcohol Precipitate and Content Analysis of Scrophulariae Radix Polysaccharides

作　　者：张婷 孙红梅 李振 魏桂杰 王宏雅 魏锋[3] 张玉杰[1] ZHANG Ting;SUN Hong-mei;LI Zhen;WEI Gui-jie;WANG Hong-ya;WEI Feng;ZHANG Yu-jie(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Infinitus (China),Guanhgzhou 529100,China;National Institutes for Food and Drug Control,Beijing 100053,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]无限极(中国)有限公司,广东广州529100 [3]中国食品药品检定研究院,北京100053

出　　处：《中国现代中药》2021年第4期679-684,697,共7页Modern Chinese Medicine

摘　　要：目的:优化水提醇沉法提取纯化玄参多糖的工艺条件,建立多糖酸水解、1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生化高效液相色谱法(HPLC)测定玄参多糖单糖组成和含量的方法。方法:在单因素试验基础上,选择溶媒量(A)、提取时间(B)和醇沉浓度(C)为考察因素,应用Box-Behnken响应面中心组合法进行三因素三水平设计,以多糖含量为指标,优化玄参多糖的提取工艺;玄参多糖用三氟乙酸(TFA)水解后,以PMP对水解单糖进行衍生化,并建立6种单糖的HPLC分离和含量测定方法。结果:玄参多糖的最佳提取工艺条件为50倍溶媒量,提取时间1 h,醇沉浓度90%;玄参多糖由甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖组成;各单糖的平均质量比为0.48∶1.40∶1.00∶69.80∶93.00∶2.20;玄参多糖平均质量分数为16.80%。结论:优选的玄参多糖水提醇沉提取纯化工艺可将玄参中的多糖提取完全,用于后续分析;PMP柱前衍生化HPLC单糖测定法的精密度、重复性和准确性良好,适用于玄参多糖的组成及含量测定。Objective:Water extraction and alcohol precipitation were used to extract and purify Scrophulariae Radix polysaccharide(RSP),and the extraction and purification conditions were optimized.A HPLC method was established for the determination of monosaccharide composition and content of RSP by acid hydrolysis and pre-column derivatization of 1-phenyl-3-methyl-5-pyrazolinone(PMP).Methods:On the basis of single factor experiment,the solvent amount,extraction time and alcohol concentration were selected as the investigation factors,Box-Behnken central combination was used for three-factor and three-level design,and response surface method was used to optimize the extraction process of RSP;After hydrolysis with trifluoroacetic acid,monosaccharides were derived by PMP,and six common monosaccharides were separated by HPLC chromatography.Results:The optimum extraction conditions of RSP were 50 times the amount of solvent,1 h extraction time and 90%ethanol concentration;RSP is composed of mannose,rhamnose,galacturonic acid,glucose,galactose and arabinose;the average quality ratio of monosaccharide was 0.48∶1.40∶1.00∶69.80∶93.00∶2.20;the average content of RSP was 16.80%.Conclusion:The optimized extraction and purification technology can completely extract the polysaccharides from Scrophulariae Radix for subsequent analysis.HPLC for precolumn derivatization of PMP has good reproducibility and stability,and is suitable for the determination of polysaccharides.

关 键 词：玄参多糖 响应面法 柱前衍生化 高效液相色谱法 单糖组成 含量 

分 类 号：R284.2[医药卫生—中药学]