雪菊黄酮有效部位化学成分与药材质量研究

发布时间：2018-06-13 14:02

  本文选题：雪菊 + 黄酮　； 参考：《北京中医药大学》2017年硕士论文

【摘要】：目的:对雪菊中黄酮类化合物进行分离纯化及结构鉴定;研究雪菊黄酮部位和药材中主要成分的含量,及药材中主要成分的含量与性状特征的相关性;通过对黄酮类化合物的降糖活性的研究,探寻雪菊药效物质基础,为质量标准建立奠定基础。方法:使用硅胶、SephadexLH-20、RP-18反向硅胶等多种柱色谱与Semi-PHPLC技术,并结合重结晶等方法,从雪菊提取物中分离黄酮类化合物,通过1M-NMR、13C-NMR、HMQC、MS等技术鉴定化合物结构;采用HPLC-DAD技术对雪菊黄酮部位及药材中的主要成分进行含量测定,并建立雪菊药材HPLC图谱,对10批雪菊样品HPLC图谱进行相似度评价;观察雪菊舌状花的特征,依据两色舌状花的红褐色部分的长度比例及颜色进行分级;以舌状花特征等级与主要成分含量为自变量,进行聚类分析,将10批样品进行分组,分析雪菊中主要黄酮类成分的含量与雪菊舌状花颜色特征的相关性;以小鼠3T3-L1前脂肪细胞为研究载体,采用葡萄糖氧化酶法考察雪菊黄酮类成分对细胞葡萄糖代谢产生的影响,给药量设计为2.5、5、10μg.ml-1。结合化学、性状与药理研究,阐明雪菊黄酮类成分与药材质量的相关性,为质量标准建立奠定基础。结果:本研究共分离得到12个化合物,鉴定出其中8个化合物的结构,分别是2S',5',5-三羟基二氢黄酮-7-O-β-D-吡喃葡萄糖苷、3',5'二羟基二氢黄酮-7-O-β-D-吡喃葡萄糖苷、2S-3',5,5',7-四羟基二氢黄酮、2S-3',4',7,8-四羟基二氢黄酮、马里苷、奥卡宁、紫铆花素-4'-O-β-D-吡喃葡萄糖苷、槲皮素-7-O-β-D-吡喃葡萄糖苷。对得到的马里苷、奥卡宁、2S-3',4',7,8-四羟基二氢黄酮、2S-3',5',5-三羟基二氢黄酮-7-O-β-D-吡喃葡萄糖苷4个主要成分在雪菊黄酮部位中的含量进行测定,测定结果为2S-3',4',7,8-四羟基二氢黄酮的含量为0.053 71mn·mg-1、2S-3',5',5-三羟基二氢黄酮-7-O-β-D-吡喃葡萄糖苷的含量为0.032 12 mg·ng-1、马里苷的含量为0.118 8 mg·mg-1、奥卡宁的含量为0.047 37 mn·mg-1,其中马里苷的含量最高。根据雪菊药材HPLC图谱的相似度分析,发现市购10批雪菊药材样品的HPLC图谱相似度均≥0.95。性状研究发现市购不同批次样品间舌状花的性状存在差异。含量测定结果表明不同批次样品的马里苷与奥卡宁含量差异较大。将雪菊舌状花红褐色部分的长度、颜色深浅及马里苷、奥卡宁含量作为自变量进行聚类分析,发现马里苷含量与红褐色部分颜色及面积存在正相关。药理研究结果为雪菊黄酮部位及各单体成分会对正常小鼠3T3-L1细胞的糖代谢产生影响,但与对照组之间不存在显著性差异。结论:雪菊黄酮部位及药材主要成分为查尔酮和二氢黄酮,其中马里苷含量最高并与药材舌状花的性状特征存在相关性,建议作为雪菊质量评价的指标性成分。
[Abstract]:Objective: to study the content of flavonoids and the correlation between the content of flavonoids and the characteristics of characters. Through the study on the hypoglycemic activity of flavonoids, the substance foundation of the medicinal effect of Xueju was explored, which laid a foundation for the establishment of quality standard. Methods: Sephadex LH-20 RP-18 reverse silica gel column chromatography and semi-PHPLC were used to isolate flavonoids from the extract of Fructus asiatica. The structures of the compounds were identified by 1M-NMR-13C-NMR-HMQCMS. HPLC-DAD technique was used to determine the contents of flavonoids and the main components in the medicinal materials. HPLC chromatograms were established to evaluate the similarity of the HPLC profiles of 10 batches of snow chrysanthemum samples, and to observe the characteristics of the tongue flowers of snow chrysanthemum. According to the length ratio and color of the reddish brown part of the two-color glossy flower, the characteristic grade and the main component content of the glossy flower were used as independent variables, and the 10 batches of samples were grouped. The relationship between the content of flavonoids in chrysanthemum and the color characteristics of chrysanthemum tongue flower was analyzed, and the effect of flavonoids on glucose metabolism was investigated by glucose oxidase method using 3T3-L1 preadipocytes in mice as the carrier. The dosage was designed to be 2.5 渭 g 路ml ~ (-1), 10 渭 g 路ml ~ (-1). The correlation between flavonoids and the quality of medicinal materials was clarified by combining chemical, character and pharmacological studies, which laid a foundation for the establishment of quality standard. Results: in this study, 12 compounds were isolated and 8 compounds were identified. The results were as follows: (2) Spline 5H _ (5) -trihydroxy dihydroflavone -7-O- 尾 -Dpyranoside 5 '-dihydroxy dihydroflavone -7-O- 尾 -D- pyranoside (2S-3) -7S-38-tetrahydroxy dihydroxy dihydrogenone, Marinine, Okanine, Violotenin-4- O- 尾 -D- glucopyranoside, Glucopyranoside, and pyrrolidine, respectively, were 2S-378-tetrahydroxy dihydroxy dihydroflavone, Marinine, Okanin, Violanol-4- O- 尾 -D- glucopyranoside, pyrrolycoside. Quercetin -7-O- 尾 -Dpyranoside. The content of the four main components of marinine, Okaninine 2S-3, 4-dihydroflavone, 7-O- 尾 -Dpyranoside, was determined in the flavonoid fraction of Schizea chinensis, and the contents of the four main components were determined in the flavonoid fraction of Cassia chinensis, and the contents of the four main components were determined in the flavonoids of Cassia chinensis, and the contents of the four main components were determined. The results showed that the content of 2S-3O4H7H7HD-THDH was 0.053 71mn mg-1mg-1and 2S-3S-3, the content of 5-trihydroxy dihydroflavone was 0.032 12 mg ng-1, the content of Marinine was 0.118 8 mg -1, the content of ocanine was 0.047 37 mn mg-1, and the content of marmalinine was the highest among the three kinds of glucoside, the content of which was 0.032 12 mg 路mg -1, the content of marinine was 0.118 8 mg 路g -1, and the content of ocanine was 0.047 37 mn mg 路g -1. According to the similarity analysis of the HPLC spectra of Xueju, it was found that the similarity of HPLC spectra of 10 batches of snow chrysanthemum samples was 鈮，

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