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白花蛇舌草甾体类化学成分及其β-谷甾醇含量测定方法的研究
Studies on the Steroidal Chemical Constituents and Quantitative Determination of β-Sitosterol of Oldenlandia Diffusa(Willd.) Roxb.
【作者】 毛杏飞;
【导师】 罗佳波;
【作者基本信息】 第一军医大学 , 中药药剂学, 2002, 硕士
【摘要】 白花蛇舌草[Oldenlandia diffusa(Willd.)Roxb.]始载于《广西中药志》,为茜草科植物的带根全草。其味苦、甘、寒,入心、肝、脾三经,功效清热解毒,清热利湿。民间常用此药治疗咽喉肿痛、癌肿等。现代药理表明白花蛇舌草具有抗菌、消炎和抗肿瘤作用,其中β-谷甾醇为白花蛇舌草中抗肿瘤有效成分之一。 实验目的:对白花蛇舌草中甾体类化学成分进行提取、分离和鉴定,并首次用薄层扫描法对白花蛇舌草中的β-谷甾醇含量测定进行研究。 实验方法: 提取 将白花蛇舌草粉碎成粗粉,加氯仿,加热回流提取三次,滤过;合并氯仿提取液,减压回收氯仿,浓缩至稠膏状,置冰箱,备用。 分离 将白花蛇舌草提取物加少量氯仿溶解,用适量的硅胶粉制样。样品与吸附剂按1:80的比例上硅胶柱,用石油醚-丙酮为洗脱剂,进行梯度洗脱,分段收集,浓缩、结晶 分别得到化合物A和B。 鉴定 化合物A、B分别经熔点、红外、核磁共振、质谱等鉴定,证明为豆甾醇和β-谷甾醇。 含量测定 精密称取干燥至恒重的白花蛇舌草5克,置索氏提取器中,加适量氯仿加热回流提取6小时,至回流液中无β-谷甾醇(薄层检测)。加1%氢氧化钠溶液150ml洗涤二次,以除去酸性杂质;再用0.5%盐酸溶液150ml洗涤一次,除去碱性杂质;最后用蒸馏水洗去残留盐酸。合并洗涤液,用适量氯仿萃取。将氯仿萃取液与氯仿提取液合并,减压回收氯仿,浓缩至适量,再用氯仿定容至10ml,以β-谷甾醇作为对照品,用薄层扫描法测定白花蛇舌草中β-谷甾醇的含量。 实验结果:白花蛇舌草粗粉用氯仿加热回流提取后,上硅胶柱,以石油醚-丙酮为洗脱剂进行梯度洗脱,洗脱液经浓缩、结晶,得化合物胆甾醇和β-谷甾醇;并以β-谷甾醇作为对照品,用薄层扫描法测定了不同批次的白花蛇舌草中β-谷甾醇的含量,其范围为0.09~0.20%。 结论:将白花蛇舌草用氯仿提取,经硅胶柱层析,可分离得到纯度较高的胆甾醇和β-谷甾醇;并可用薄层扫描法测定白花蛇舌草中的的β-谷甾醇含量,此法简单准确,重现性好,回收率高,结果稳定,为白花蛇舌草的质量研究提供了方法和依据。
【Abstract】 Oldenlandia diffusa( Willd.)Roxb. is recorded in the <<Guangxi Chinese herbal medicine ambition>> for the first time .The herbal is a whole grass plant of Rubiaceae.Property,flavour and channel tropism:bitter and sweet in flavour,cool and cold in property,acting on the heart,liver,spleen channel.Effects:heat-clearing and detoxicating,heat-clearing and inducing dampness.In folks,this medicine can be used to treat commonly swollen and sore throat,cancer ... etc..The modern pharmacology expresses that the herbal medicine can be antibiosis,eliminate inflamation and anticarcinogenic,and P-Sitosterol is one of active constituents of Oldenlandia diffusa( Willd.)Roxb.. Objective:to study on extraction and separation and identified of the steroidal chemical constituents,and to establish the method of quantitative determination of P-Sitosterol in Oldenlandia diffusa(Willd.) Roxb.Methods:the constituents were gained by refluxed with CHC13 for three times,then were separated and puried by column chromatography with silica gel.The strutures were identified by mp. IR. NMR. MS. Quantitaty of P-Sitosterol in Oldenlandia diffusa(Willd. ) Roxb. was determined by TLC-scanning for the first time.Results:Two compounds were obtained from Oldenlandia diffusa(Willd. ) Roxb extraction and identified as Stigmasterol (A),P -Sitosterol (B). Quantitaties of P-Sitosterol in Oldenlandia diffusa(Willd.) Roxb. were 0.09-0.20% respectively.Conclusion:The method was highly sensitive reproducible for quantitative determination of P-Sitosterol. The method can be used for the quality control of Oldenlandia diffusa(Willd.) Roxb.. Two compounds can be gained from Oldenlandia diffusa(Willd.) Roxb and identified as Stigmasterol (A),0 -Sitosterol (B).
【Key words】 Oldenlandia diffusa(Willd.)Roxb; Extraction and separation; silica gel column; NMR; MS; IR; Stigmasterol; β -Sitosterol; TCL-scanning;
- 【网络出版投稿人】 第一军医大学 【网络出版年期】2003年 01期
- 【分类号】R284
- 【被引频次】8
- 【下载频次】792