【作者】 郅妙利；

【导师】 许启泰； 康文艺；

【作者基本信息】 河南大学 ， 中药学， 2008， 硕士

【摘要】 本论文研究了茜草中RMC和QA的提取分离、结构鉴定和含量测定等内容,并确立了连翘中芦丁和卷柏中穗花衫双黄酮的含量测定方法。对RMC的研究:首先以提取溶剂、料液比、提取时间、提取次数4个因素,每因素设3个水平,RMC提取率(%)为评价指标进行L9(34)正交实验,确定了RMC最佳提取工艺为20倍量石油醚提取2次,每次2个小时,提取率为0.8%;然后依次用常压硅胶柱、凝胶柱和减压硅胶柱色谱分离手段等进行分离,用重结晶法进行纯化;得到的化合物用MS、1H NMR和13C NMR等波谱学方法进行结构鉴定;用反相高效液相色谱法(RP-HPLC)测定了栾川、四川、湖北、广西、贵州、河南(百氏康)和陕西7个产地的中药材中RMC含量。色谱柱为Purospher star(250 mm×4.6 mm,5μm, Merk公司),流动相为甲醇-水-四氢呋喃(90:9.3:0.7),检测波长为250 nm,流速为1.0 mL/min,柱温为25℃。结果RMC进样量与峰面积在0.200～2.00μg范围内线性关系良好(r=1)。除陕西药材分离效果不好外,其它6产地茜草中RMC含量(%)为栾川(1.23%)>四川(0.77 %)>湖北(0.75 %)>广西(0.60 %)>贵州(0.37 %)>河南(百氏康)(0.32 %)。对QA的研究:茜草药材用丙酮-水(7:3)冷浸提取,得到的浸膏依次用石油醚、乙酸乙酯和正丁醇萃取,萃取得到的正丁醇部分依次用常压硅胶柱、加压硅胶柱、凝胶柱、减压硅胶柱等色谱分离手段进行分离,最后用重结晶法进行纯化;采用MS、1H NMR、13C NMR等波谱学方法进行结构鉴定;使用RP-HPLC法测定栾川、陕西、广西、四川、贵州、湖北6产地药材中QA含量,流动相为甲醇-水-四氢呋喃(65:34.7:0.3),流速为1.0 mL /min,检测波长为276 nm,柱温为25℃,外标法定量分析。结果表明进样量与峰面积在0.020μg～0.160μg之间时,线性关系良好(r=0.9998)。陕西和栾川样中QA含量分别为0.053 %和0.42 %,其它产地药材可能因其成分复杂而导致分离效果不佳。连翘中芦丁的含量测定:用RP-HPLC法测定连翘果实、7月叶、7月茎、老叶中芦丁的含量,流动相为甲醇-2%冰乙酸水溶液(42:58),流速为0.8 mL /min,检测波长为359 nm,柱温为25℃。该方法的线性范围为0.68～6.8μg,r = 1,平均回收率为98.0 %,RSD为2.0% ( n = 6)。结果测得,连翘果实、7月叶、7月茎和老叶所含芦丁的量为:7月叶(0.92%)>老叶(0.35%)>果实(0.28%)>7月茎(0.12%)。卷柏中穗花衫双黄酮的含量测定:用RP-HPLC测定垫状卷柏和卷柏药材中的穗花杉双黄酮的含量,流动相为甲醇-0.1%磷酸水溶液(65:35),流速为0.8 mL /min,检测波长为337 nm,柱温为25℃。该方法的线性范围为0.038～0.342μg,r = 0.9999。实验测得贵州和湖南卷柏中穗花杉双黄酮含量分别为0.19%和0.51%,西藏产垫状卷柏所含穗花杉双黄酮含量为0.36%。更多还原

【Abstract】 The paper studied the extraction, isolation, identification and content determination of RMC and QA from Rubia cordifolia L., and established the method of content determination of rutin and amentoflavone from Forsythia suspense and Selaginella tamariscina(Beauv.)Spring respectively.Studies on RMC: material ratio and the weight of dry rubia material (V/W) and time have been investigated for their influences on the extraction rate (%) of RMC. Oputimum combinations of extraction conditions were obtained using the L9(34) orthogonal analysis technique. The result showed that the material ratio had the most evident influence. It was found that an extraction rate of 0.8 % could be achieved at petroleum. A material ratio of 20:1 and an extraction time of 2 h and 2 times were efficacy.All kinds of silica gel column chromatography including nomal pressure and decompression and Sephadex LH-20 chromatography were used to isolate RMC in a sequence order. MS, 1H NMR and 13C-NMR were used to identify the structure of RMC. RP-HPLC (reversed-phase high-performance liquid chromatography) method was used to determine the content of RMC. Seven samples from seven regions including Luanchuan, Sichuan, Hubei, Guangxi, Guizhou, Henan (baishikang) and Shanxi. HPLC analysis conditions were the Purospher star column (250 mm×4.6 mm, 5μm, Merk) at 25℃using methanol-water-Tetrafuran (90:9.3:0.7) as mobile phase with a flow rate of 1.0 ml/min and detection wavelength at 250 nm. There was good linear relationship between the peak area and the sample content injected at the ranges of 0.200～2.00μg (r=1) for RMC. The result showed that the content of RMC for different samples were 1.23 % (Luanchuan), 0.77% (Sichuan), 0.75% (Hubei), 0.60 % (Guangxi), 0.37 % (Guizhou) and 0.32 % ( Henan baishikang). The sample from Shanxi had a shoulder peak and couldn’t be quantified.Studies on QA: the roots of R. cordifolia were extracted using acetone-water (7:3, V/V). After evaporation of the solution, the residue was suspended in water and extracted with petroleum, ethyl acetate and n-butanol. The n-butanol extract was isolated for QA.All kinds of silica gel column chromatography including nomal pressure and decompression and Sephadex LH-20 chromatography were used to isolate QA in a sequence order. MS, 1H NMR and 13C-NMR were used to identify the structure of RMC. RP-HPLC (reversed-phase high-performance liquid chromatography) method was used to determine the content of QA. Six samples from six regions including Luanchuan, Sichuan, Hubei, Guangxi, Guizhou and Shanxi. HPLC analysis conditions were the Purospher star column (250 mm×4.6 mm, 5μm, Merk) at 25℃using methanol-water-tetrafuran (65:34.7:0.3) as mobile phase with a flow rate of 1.0 ml/min and detection wavelength at 276 nm. There was good linear relationship between the peak area and the sample content injected at the ranges of 0.020～0.160μg (r=0.9998) for QA. The result showed that the content of QA for different samples were 0.42 % (Luanchuan), 0.053 % (Shanxi). QA in other samples from other regions couldn’t be seperated well.Content determination of rutin: RP-HPLC (reversed-phase high-performance liquid chromatography) method was used to determine the content of rutin extracted from F. suspensa. HPLC conditions were the Purospher star column (250 mm×4.6 mm, 5μm, Merk) at 25℃using methanol-2% glacial acetic acid (42:58) as mobile phase with a flow rate of 0.8 ml/min and detection wavelength at 359 nm. There was good linear relationship between the peak area and the sample content injected at the ranges of 0.68～6.8μg (r=1) for rutin.Content determination of amentoflavone: RP-HPLC (reversed-phase high-performance liquid chromatography) method was used to determine the content of amentoflavone from S. tamariscina(Beauv.)Spring and S. pulvinate(Hook.Et Grev.)Maxim. HPLC conditions were the Purospher star column (250 mm×4.6 mm, 5μm, Merk) at 25℃using methanol- 0.1% phosphoric acid (65:35) as mobile phase with a flow rate of 0.8 ml/min and detection wavelength at 337 nm. There was good linear relationship between the peak area and the sample content injected at the ranges of 0.038～0.342μg (r=0.9999) for amentoflavone.更多还原