川产中柴2号主要化学成分及指纹图谱研究

发布时间：2018-04-30 00:43

本文选题：中柴2号 + 柴胡皂苷　；参考：《西南科技大学》2017年硕士论文

【摘要】：柴胡作为我国传统常用中药,药用历史悠久。在2015年版的《中华人民共和国药典》中收录的柴胡原植物有两种,即北柴胡(Bupleurum chinense DC.)和狭叶柴胡(Bupleurum scorzonerifolium Willd.)。柴胡苦辛微寒,具有解热、退热、抗菌、抗病毒、解除胸闷胀痛的作用。柴胡属植物广泛分布在我国各地,除海南省未见报道外,其他各省均有柴胡属植物分布,近些年来,四川广元和绵阳已逐渐成为柴胡的主要产地。本研究分析了川产北柴胡中柴2号中柴胡皂苷、黄酮、多糖等成分含量并建立中柴2号的HPLC和GC指纹图谱,为中柴2号药材质量评价、品种鉴别及其入药提供科学依据。主要结果如下:通过对中柴2号样品预处理条件、检测条件、洗脱条件的优化,流动相、色谱柱的选择,确定了柴胡皂苷与黄酮类成分测定的最终色谱条件为Hypesil GOLD色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈(A)-0.1%磷酸水溶液(B);梯度洗脱程序(0~5 min,10%A~30%A;5 min~25 min,30%A~70%A);流速为0.8 m L·min-1;柱温检测为常温(30oC),进样量10μL,运行时间25 min,检测波长210 nm。采用HPLC法测定中柴2号中皂苷与黄酮类成分,结果发现中柴2号地下部分中含有柴胡皂苷a、柴胡皂苷c和柴胡皂苷d,且柴胡皂苷a和柴胡皂苷d含量总和均大于0.30%,高于2015年版《中国药典》柴胡药材含量测定项下的最低限度规定,最高可达1.01%;地下部分仅含有少量的黄酮类成分。中柴2号地上部分中含有少量柴胡皂苷a和柴胡皂苷d,其和最高为0.10%;地上部分含有较多的黄酮类成分,其含量最高为0.68%。采用紫外-可见分光光度法测定中柴2号中总皂苷、总黄酮和多糖含量,结果发现中柴2号地上部分柴胡总皂苷含量为0.26%~0.82%,总黄酮含量为0.81%~2.76%,可溶性多糖含量为0.996~3.859 mg·g-1,粗多糖含量为6.032~10.632 mg·g-1,总多糖含量最高可达12.837 mg·g-1。地下部分柴胡总皂苷含量为1.05%~2.12%,总黄酮含量为0.18%~0.46%,可溶性多糖含量为2.118~13.809 mg·g-1,粗多糖含量为7.153~20.257 mg·g-1,总多糖含量最高可达28.992 mg·g-1。中柴2号地上部分总皂苷含量在8、9月份达到最高值,10、11月份总皂苷含量降低,地下部分在总皂苷含量在7、8月份达到最高值,9、10、11月后总皂苷含量逐渐降低;中柴2号地上部分总黄酮含量在8、9月份达到最高值,10、11月份逐渐降低,中柴2号地下部分总黄酮含量在7、8月份达到最高值,9、10、11月后逐渐降低;中柴2号地上部分及地下部分总多糖含量从7月份至11月份基本呈上升趋势。10批中柴2号样品HPLC及GC色谱图导入《中药色谱指纹图谱相似度评价系统》,HPLC指纹图谱匹配10个共有峰并确定柴胡皂苷a、柴胡皂苷c和柴胡皂苷d三个有效成分,共有峰相对保留时间RSD值在0.18%~0.70%之间,共有峰相对峰面积RSD值在20.95%~101.05%之间;GC指纹图谱匹配12个共有峰,共有峰相对保留时间RSD值在0.04%~0.32%之间,共有峰相对峰面积RSD值在20.67%~88.92%之间,结果表明10批柴胡主要成分基本相同,但各成分含量差异较大,即相同品种不同产区川产柴胡各化学成分的含量有明显差别。中柴2号HPLC指纹图谱相似度结果表明各批柴胡样品与对照指纹图谱的相似度在0.39%~0.97%之间,其它各样品间的相似度在0.05%~0.95%之间;中柴2号GC指纹图谱相似度结果表明各批柴胡样品与对照指纹图谱的相似度在0.76%~0.99%之间,其它各样品间的相似度在0.33%~0.98%之间。虽然各批柴胡样品基源相同,但由于生长环境不同,指纹图谱相似度也有所不同。本课题通过对四川种植基地10批柴胡中柴2号指纹图谱的研究,建立了川产柴胡中柴2号HPLC与GC指纹图谱分析方法,将中柴2号柴胡皂苷成分与挥发油成分的共有峰信息分别在GC与HPLC图谱中展现,更全面科学的对药材质量进行评价,该方法简单易行,重现性好,为川产柴胡药材的品质监控提供了一定的科学依据。
[Abstract]:Bupleurum Bupleurum is a traditional Chinese traditional medicine with a long history of medicinal use. In the 2015 edition of the People's Republic of China Pharmacopoeia, there are two kinds of Bupleurum bupleurum, which are Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd.. The bupleurum Bupleurum is bitter and cold, it has antipyretic, antipyretic, antiviral, antiviral, and relieving pain in the chest. The genus Bupleurum is widely distributed in all parts of China, except in Hainan province. In other provinces, the genus Bupleurum is distributed. In recent years, Sichuan Guangyuan and Mianyang have gradually become the main producing areas of Bupleurum Bupleurum. This study analyzed the content of Bupleurum bupleurum, flavonoids and polysaccharides in Bupleurum Bupleuri No. 2 in Sichuan Province, and set up No. 2 of Zhong Chai. The HPLC and GC fingerprints provide scientific basis for the quality evaluation of Chinese Chai 2, variety identification and drug delivery. The main results are as follows: the final chromatographic conditions for the determination of saponins and flavonoids of Bupleurum saponins and flavonoids are determined to be Hypesi by the pretreatment conditions of the sample of Zhong Chai No. 2, the conditions of detection, the optimization of the elution conditions, the selection of the mobile phase and the chromatographic column. The L GOLD column (4.6 mm x 250 mm, 5 mu m), the mobile phase is acetonitrile (A) -0.1% phosphoric acid solution (B), the gradient elution program (0~5 min, 10%A~30%A, 5 min~25), the flow velocity is 0.8, and the column temperature is measured as normal temperature, the sample volume is 10 mu, the transportation time 25, and the detection wavelength 210 for the determination of saponins and flavonoids in the middle Chai 2 The results found that the underground part of Zhong Chai 2 contains saponins a, bupleurum saponins C and Bupleurum saponins D, and the total content of Radix Bupleurum saponins A and Radix Bupleurum saponins D is more than 0.30%, which is higher than the minimum regulation of Chinese Pharmacopoeia < Chinese Pharmacopoeia > Radix Bupleuri content determination, up to 1.01%. The underground part only contains a small amount of flavonoids. The upper part of Zhong Chai No. 2 contains a small amount of Bupleurum saponins A and Bupleurum saponins D, and the highest is 0.10%; the upper part of the ground contains more flavonoids. The highest content is 0.68%. using UV visible spectrophotometry to determine the total saponins, total flavones and polysaccharides content in No. 2 of Zhong Chai, and the results found the total saponins content of Bupleurum Bupleurum in the middle part of Chai 2. The content of total flavonoids was 0.81%~2.76%, the content of soluble polysaccharide was 0.996~3.859 mg. G-1, the content of crude polysaccharide was 6.032~10.632 mg. G-1, the total polysaccharide content was up to 12.837 mg. G-1., the total content of total saponins of Radix Bupleuri was 1.05%~2.12%, the content of total flavonoids was 0.18%~ 0.46%, and the content of soluble polysaccharide was much more coarse. The content of sugar was 7.153~20.257 mg g-1, the content of total polysaccharide was up to 28.992 mg. G-1., the total saponsides content of Chai No. 2 reached the highest value in the month of 8,9, the total saponins content in the month of 10,11 decreased, the total saponins content in the underground part reached the highest value in 7,8 month, and the total saponins content gradually decreased after 9,10,11 month, and the upper part of Zhong Chai 2 was total. The content of flavonoids reached the highest value in the month of 8,9 and gradually decreased in the month of 10,11. The content of total flavonoids in the underground part of Zhong Chai 2 reached the highest value in the month of 7,8, and gradually decreased after 9,10,11 months. The total polysaccharide content of the upper and underground parts of the middle Chai No. 2 was basically up from July to November. 10 common peaks were matched by HPLC fingerprints and three effective components of Bupleurum saponins a, bupleurum saponins C and Bupleurum saponins D, the RSD value of the total peak relative retention time was between 0.18%~0.70%, the total peak area RSD value was between 20.95%~ 101.05% and the GC fingerprint matched 12 common peaks. The peak relative retention time RSD value was between 0.04%~0.32%, and the total peak relative peak area RSD value was between 20.67%~88.92%. The results showed that the main components of the 10 batches of Radix Bupleuri were basically the same, but the content of each component was different, that is, the content of the chemical composition of the radix Bupleuri in the same variety of different producing areas was distincently different. The similarity of the HPLC fingerprint of Zhong Chai 2 was similar. The results showed that the similarity degree of each batch of Radix Bupleuri samples and the control fingerprints was between 0.39%~0.97%, the similarity of all kinds of products was between 0.05%~0.95%, and the similarity of GC fingerprint of Zhong Chai 2 showed that the similarity degree of each batch of Radix Bupleuri samples and the control fingerprints was between 0.76%~0.99% and the similarity of all kinds of products was in 0.33%~0.98%. Although the base source of all batches of Radix Bupleuri was the same, the similarity of fingerprints was different because of the different growth environment. Through the study on the fingerprint of 10 batches of Chaihu Zhong Chai 2 in Sichuan planting base, the method of HPLC and GC fingerprint analysis of Bupleurum chinchai 2 was established, and the composition of Bupleurum saponins and the volatile oil composition of Zhong Chai 2 The common peak information was displayed in the GC and HPLC maps, and the quality of the medicinal materials was evaluated in a more comprehensive and scientific way. The method was simple and reproducible, which provided a scientific basis for the quality control of Radix Bupleuri in Sichuan Province.

【学位授予单位】：西南科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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