5-9星点设计效应面法优化白茅根总酚酸的提取工艺-.doc

星点设计-效应面法优化白茅根总酚酸提取工艺刘荣华基金项目 国家自然科学基金资助项目(No.30860363)作者简介 刘荣华（1964-），男（汉族），教授，理学博士，从事中药质量评价和中药活性物质基础研究。Tel：(0791)7119010，E-mail：，陈石生，任刚，邵峰，黄慧莲，杨明(江西中医学院现代中药制剂教育部重点实验室，江西 南昌 330004)摘要 目的：优化白茅根总酚酸的提取工艺。方法：采用超声辅助提取法，选取提取试剂浓度、提取时间、提取温度和料液比作单因素试验，在此基础上采用星点设计考察提取试剂浓度、提取时间及提取温度3因素对提取工艺的影响，对结果进行二项式方程拟合，效应面法筛选最佳条件并进行预测分析。结果：得到的最佳提取工艺条件为：乙醇浓度为40%，提取时间38min，提取温度为33 ，料液比为1:40，总酚酸含量的预测值为2.7102 mgg-1。验证实验的总酚酸含量为2.4625 mgg-1，与预测值接近，说明模型可靠。结论：该工艺步骤简单，稳定可行，为白茅根进一步的研究提供科学依据。关键词 白茅根；总酚酸；提取工艺；星点设计；效应面法Optimal Extraction of Total Phenolics from the Roots of Imperata cylindrica Based on Central Composite Design and Response Surface MethodologyLIU Rong-hua*, CHEN Shi-sheng, REN Gang, SHAO Feng, HUANG Hui-lian, YANG Ming(Key Laboratory of Modern Preparation of TCM, Jiangxi University of TCM, Ministry of Education, Nanchang 330004, China)ABSTRACT AIM: To identify the optimal conditions of extraction technology of total phenolics in Rhizoma Imperatae. METHODS: The influencing factors were studied by the single factor experiment, and central composite design and response surface methodology were applied to optimize the extraction process for total phenolics in terms of the content of total phenolics. RESULTS: The optimum extraction conditions were as follows: 40.00% ethanol as extraction solvent at 32.86 for 37.81 min, and the ratio of liquid to material was 1:40. Under these conditions, the extraction rate was 2.7102 mgg-1. CONCLUSION: The process is simple, provided high extraction rate, To provide a scientific basis for further research of Rhizoma Imperatae. KEY WORDS Imperata cylindrica; Total Phenolics; Extraction Technology; Central Composite Design; Response Surface Methodology白茅根为禾本科植物白茅Imperata cylindrica Beauv. Var. major(Nees) C. E. Hubb. 的干燥根茎，又名兰根、地筋、白花茅根、茅草根、甜草根等，广泛分布于全国各地，具有凉血止血、清热生津、利尿通淋之功效1。白茅根酚酸类成分有Tricin、Jaceidin、Quercetagetin-3,5,6,3-tetramethyl ether、3,5-Di-O-methyl-kaempferol、3-Hydroxy-4-methoxy benzaldehyde，Impecyloside，Cylindol A、B，graminones A、B，Imperanene，5-hydroxy-2-(2-phenylethyl)chromone、5-hydroxy-2-styrylchromone、5-hydroxy-2-2-(2-hydroxyphenyl)ethylchromone、flidersiachromone，p-Coumaric acid2-8。药理研究显示白茅根中酚酸类成分具有血管舒张、血小板聚集抑制活性、抗氧化活性、细胞毒性、p-糖蛋白抑制活性、糖原磷酸化酶A抑制活性、抗炎活性、抗HBV病毒复制、神经保护作用等生物活性4-12。因此，有必要对白茅根中酚酸类成分的提取工艺进行研究。本研究采用超声波辅助提取法（Ultrasound-associated extraction，UAE），在单因素考查的基础上利用星点设计(central composite design，CCD)，通过三维效应面(three dimensional response surface)进行数据处理与拟合，优化白茅根总酚酸的提取工艺，为白茅根中酚酸类成分的提取提供了依据。1 材料KQ-300DB型数控超声波清洗器(昆山市超声仪器有限公司)；UV-2550型紫外分光光度计(日本Shimadzu公司)；R-210型旋转蒸发仪(瑞士Buchi公司)；Sartorious BT 124S型万分之一天平(赛多利斯科学仪器(北京)有限公司)；GZX-9070MBE型数显鼓风干燥箱(上海博讯实业有限公司医疗设备厂)；高速中药粉碎机(瑞安市永历制药机械有限公司)；S-4药典检验筛(浙江上虞市华康化验仪器厂)。白茅根于2010年3月采于江西中医学院湾里校区，经江西中医学院赖学文教授鉴定为禾本科植物白茅Imperata cylindrica Beauv. Var. major(Nees) C. E. Hubb.的根茎，凭证标本存于江西中医学院标本馆。没食子酸对照品(中国药品生物制品检定所，批号110831-200803)，铁氰化钾、三氯化铁、盐酸、无水乙醇均为分析纯，双蒸水自制。2 方法与结果(略)3. 讨论超声波辅助提取法(UAE)是利用超声空化效应，同时提供机械作用使溶剂更好的渗透进药材基体，增加固液两相的接触面积，加速植物有效成分的浸出提取14,15。该法所需设备简单，操作方便，比一般提取技术更加有效16，超声过程中因为化学参与及提取时间的减少，故可以防止目标成分可能的化学降解17，而温度过高会加速酚性成分的氧化分解，所以UAE更加适合酚酸类成分的提取。星点设计(CCD)是在析因设计的基础上，加上星点及中心点而形成的实验次数较少、可采用三维效应面(three dimensional response surface)进行数据处理与拟合的实验设计方法18，已在药物提取和处方工艺的优化中有一定的应用19-23。本实验在单因素试验的基础上，利用星点设计-效应面法优化白茅根总酚酸的提取工艺，可为白茅根中酚酸类成分的提取提供依据，这对于进一步研究白茅根酚酸类成分具有重要意义。由单因素试验和星点试验设计及其分析结果获得的超声辅助提取白茅根总酚酸的最佳工艺为乙醇浓度为40%，提取时间38 min，提取温度为33 ，料液比为1:40。此时总酚酸得率为2.4625 mgg-1。本研究结果是在实验室小试基础上获得的，还有待在放大实验中进一步验证。参考文献1 国家中医药管理局.中华本草,第八册M.上海科学技术出版社, 357-3602 Gamal A. Mohamed, Ahmed Abdel-Lateff, Mostafa A. Fouad,et al. Chemical Composition and Hepato-protective activity of Imperata cylindrica Beauv.J. Pharmacognosy Magazine, 2009, 4(17): 28-36.3 Lee, Dae-Young, Han, Kyung-Min, Song, Myoung-Chong, et al. 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