金钗石斛总生物碱提取及含量测定方法的改进  

The extraction and the improvement of determination method for total alkaloids in Dendrobium nobile Lindl

作　　者：赵福坤[1] 吴芹[1] 陆远富[1] 杨文进[1] 姚晓东[2] 张茂生[2] 石京山[1] 

机构地区：[1]遵义医学院药理学教研室暨基础药理省部共建教育部重点实验室,贵州遵义563099 [2]遵义医学院药学院药物化学教研室,贵州遵义563099

出　　处：《遵义医学院学报》2015年第5期532-535,共4页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:30660208;NO:30960447);贵州省教育厅"125"重大专项(NO:黔教合重大专项[2012]012)

摘　　要：目的提取较高纯度的金钗石斛总生物碱,并简单、准确地测定总生物碱含量。方法金钗石斛药材以90%乙醇10倍量浸泡12 h,然后加热煮沸2 h,连续3次。醇提液减压浓缩至无醇味后以5%盐酸溶解,抽滤,酸水溶液经石油醚萃取3次,脱脂,浓氨水碱化至p H=10,碱水液经三氯甲烷萃取2次,回收溶剂,即得生物碱粗提物。然后用氯仿-甲醇溶液(1∶1,v/v)溶解,过LH-20凝胶柱,回收溶剂,冷冻干燥,得石斛生物碱提取物。分别采用气相色谱法和酸性染料比色法测定提取物中石斛碱以及总生物碱的含量,以石斛碱(C16H25NO2)为基准物计算金钗石斛中总生物碱的含量。结果提取5kg药材,得提取物干膏22.5 g,气相色谱法测得石斛碱含量为61.6%,酸性染料比色法测得总生物碱含量为73.2%。结论LH-20凝胶柱可去除总生物碱粗提物中大量杂质,且改进后的酸性染料比色法较传统的方法更简单、准确性更高,此研究为后期的药效研究奠定了基础。Objective To extract the high purity total alkaloids of Dendrobium nobile Lindl. and determine the content accurately with simple operation. Methods Herbs were soaked with 10 times the amount of 90% ethanol for 12 h, and then heated to boiling for 2 h, 3 times in a row. The alcohol extract was concentrated with non - alcoholic, and then dissolved with 5% hydrochloric acid, leaching, aqueous acid solution was extracted 3 times by petroleum ether to de -fat, added stronger aqueous ammonia to pH 10, the alkaline solution was extracted twice with chloroform, concentrating, and the total alkaloids were obtained. Then dissolved the total alkaloid with chloroform and methanol solution （ 1： 1, v/v） to pass the LH - 20 gel column, recovered the solvent, higher purified total alkaloids were got. Determined the purity of dendrobine extracted from D. nobile Lindl. by gas chromatography, and determined the purity of total alkaloid extracted from D. nobile Lindl. by acid dye colorimetry and treated dendrobine （C16H25NO2） as the basis to calculated the content of total alkaloid. Results Extracted 5 kg herbs, and obtained the dry total alkaloid 15 g. The purity of dendrobine was 61.6% which was measured by gas chromatography, and the purity of total alkaloids was 73.2% which was measured by acid dye colorimetry. Conclusion LH -20 get column can remove impurities in crude extract of total alkaloid, and the improved acid dye colorimetry is simpler with higher accuracy than the traditional methods. This research may lay the foundation of the future study.

关 键 词：金钗石斛 石斛碱 生物碱 酸性染料比色法 气相色谱法 

分 类 号：R927.2[医药卫生—药学]