【作者】 杨继红；

【导师】 胡久梅；

【作者基本信息】 西南交通大学 ， 微生物与生化药学， 2009， 硕士

【摘要】 本研究以都江堰道地药材川芎(Ligusticum chuanxiong Hort.)为研究对象,收集了都江堰不同地区2008年6月采收的川芎样品10个、都江堰石羊镇同一川芎种植田2009年不同生长期的川芎地下部分样品12个和川芎地上部分样品11个,对其进行高效液相指纹图谱分析研究。本研究首先建立了川芎药材高效液相色谱的分析方法,色谱柱为C18(250mm×4.6mm,5μm),流动相为甲醇-1%醋酸水溶液,采用梯度洗脱,柱温为25℃,流速为1.0ml/min,检测波长为280nm。确定了7个色谱峰作为川芎药材指纹图谱特征峰,其中包括了阿魏酸的色谱峰。对都江堰石羊镇同一川芎种植田2009年不同生长期的川芎地下部分12个样品进行高效液相指纹图谱分析,并绘制川芎中阿魏酸的含量变化曲线。实验结果发现川芎有效成分含量最高的时期为小满前十天左右。川芎药材富含糖质容易虫蛀,本研究还对虫蛀川芎和正常川芎样品进行了高效液相指纹图谱分析,二者图谱比较发现虫蛀后的川芎主要成分峰值基本上都减少,有的成分甚至消失,其中有一个未知成分的含量显著增加(峰值增加),本研究命名该物质峰为“峰P”,“峰P”有可能作为川芎质量控制中虫蛀川芎指纹图谱特征峰。其次建立川芎地上部分高效液相指纹图谱的分析方法,确定了6个色谱峰作为川芎地上部分指纹图谱特征峰,该6个峰为色谱峰总面积的53.6%。说明该6个主要的共有峰能够准确的反映川芎地上部分的质量。并对同一川芎种植田采收期的紫杆川芎和绿杆川芎的地上部分以及川芎药材分别进行高效液相指纹图谱分析。通过“中药色谱指纹图谱相似度评价系统”分析发现,紫杆川芎和绿杆川芎药材相似度为0.991,紫杆和绿杆川芎地上部分的相似度也为0.991,实验结果说明紫杆川芎和绿杆川芎主要成分无明显差别,仅含量上有细小的差别。最后比较川芎地上部分和川芎药材的高效液相指纹图谱,其中相对保留时间RSD<3%,相匹配峰就达122个,其中峰面积占总峰面积3%以上的匹配峰共有19个,阿魏酸不包括在这19个峰里面,但其峰面积值相差比较大。本研究建立了都江堰川芎药材和川芎地上部分的高效液相指纹图谱分析方法,为科学评价川芎的质量作出了有意义的探索,研究结果可为川芎GAP种植基地的生产及质量控制提供一定的参考依据。更多还原

【Abstract】 Ligusticum chuanxiong Hort. in Dujiang Dam were studied.10 samples of chuanxiong in different habitats in June 2008 and 12 in different periods in 2009 were collected. And 11 samples of chuanxiong aerial parts in different periods in 2009 were collected.HPLC (High Performance Liquid Chromatogram) fingerprint and the analysis methods of genuine medicinal herbs of chuanxiong in Dujiang Dam were established. The chromatographic column of C18 (250mm×4.6mm,5μm) was used, the mobile phase was methanol-1% acetic acid solution, gradient elution was used, the temperature of chromatographic column was 25℃, the flow rate was 1.0 ml/min and the detection wavelength was at 280nm. The method showed 7 common peaks in which ferulic acid were included. Fingerprints of 12 samples in 2009 showed that the period of collection is May 10,2009 when ferulic acid and other major ingredients were the most. So the best acquisition period of chuanxiong was during May 8 and 12. Fingerprintings of samples in 2008 bitten by insect showed that their main ingredients have generally reduced, but a significant increase in concentration of a substance whose peak was called "Peak P". "Peak P"could be consider as a special peak in chuanxiong bitten by insect.The HPLC method of chuanxiong aerial parts was also established and the mobile phase was methanol-0.5% acetic acid solution.6 common peaks were shown, which were 53.6% of the total chromatographic peak area. That means the 6 common peaks can accurately reflect the quality of the aerial parts of chuanxiong. Fingerprinting showed that the similarity of green ones and purple ones reached more than 0.99, which was the same both in chuanxiong medicinal parts and aerial parts. It shows that the major components of the two species of chuanxiong were basically the same, so these two kinds of chuanxiong can be mixed.Compared the fingerprintings of chuanxiong medicinal parts with aerial parts, 122 common peaks of which the relative retention time RSD<3% were found. There are 19 peaks, which peak area is more than 3% of the total chromatographic peak area. Hoever, their peak areas are largely different. As there was little ferulic acid in aerial parts of chuanxiong, ferulic acid peaks was not included in these 19 peaks.This study established HPLC fingerprintings and analysis methods of the chuanxiong medicinal parts and aerial parts in Dujiang Dam. The method should be helpful explore as a new quality control method for traditional Chinese medieine (TCM). And also will provide some references for the GAP (Good Agricultural Practices) planting of chuanxiong.更多还原