【作者】 高璐；

【导师】 麻彤辉；

【作者基本信息】 辽宁师范大学 ， 细胞生物学， 2011， 硕士

【摘要】 桃金娘科(Myrtaceae)蒲桃属(Syzygium)植物丁香(Eugenia caryophylla Thunb.)是《中国药典》2005年版收录的中草药,其中公丁香是植物丁香药用未开放的干燥花蕾,母丁香是植物丁香的成熟果实,丁香油具有良好的抗菌消炎作用。公丁香和母丁香的主要活性成分是植物丁香不同生长时期的次生代谢产物,为阐明公丁香和母丁香药理作用差异的物质基础,本文利用高速逆流色谱法分析比较公丁香和母丁香非挥发油成分的同时,并利用高速逆流色谱和高效液相色谱技术对母丁香的化学成分进行了系统研究。本论文由三部分组成,主要包括以下几方面的研究内容:首先,综述了公丁香和母丁香化学成分、药理作用、分离提取方法的研究进展以及高速逆流色谱的应用现状与发展趋势。其次,将公丁香和母丁香95%乙醇回流提取液经逐级萃取、大孔吸附树脂进一步净化,得到公丁香和母丁香的粗提物。利用高速逆流色谱法分离公丁香与母丁香粗提物,并对其主要化学成分进行比较分析。采用溶剂体系正己烷-乙酸乙酯-甲醇-水(5:8:6:13,v/v/v/v)和(5:8:9:10,v/v/v/v)分布洗脱,公丁香和母丁香以主机转速880 r/min,流速1.2 mL/min,单次进样量70 mg各分离得到9个洗脱组分,两张高速逆流色谱分离图谱的重叠率为66.67%,其中包含6个共有峰,并且各共有峰HSCCC保留时间的相对标准偏差RSD的平均值为2.260%,与国家标准关于方法学考察资料技术参数对RSD≤3%的要求相符合,可以判定公丁香和母丁香化学成分内在质量即存在稳定性和均一性,也具有差异性,说明高速逆流色谱法比较公丁香和母丁香化学成分的可行性,为构建公丁香和母丁香高速逆流色谱指纹图谱奠定基础。同时本实验以上述条件成功地从公丁香和母丁香粗提物中分离得到槲皮素、鞣花酸、5,7-二甲氧基-2-甲基色原酮、5,7-二甲氧基-2,6-二甲基色原酮、3,5,3’4’-四羟基-7-甲氧基黄酮五种非挥发油成分,通过标准品对照以及1~H-NMR、(13)~C-NMR、MS等波谱方法对各化合物的结构进行鉴定。其中化合物5,7-二甲氧基-2,6-二甲基色原酮的结构是首次通过核磁数据进行表征。最后,在第二部分实验的基础上,利用高效液相色谱技术分离纯化母丁香强极性和弱极性粗提物,得到没食子酸、5,7,3′,4′,5′-五甲氧基黄酮、母丁香酚三种高纯度化合物,通过标准品对照以及1~H-NMR、(13)~C-NMR、MS等波谱方法对各化合物的结构进行鉴定。其中化合物5,7,3′,4′,5′-五甲氧基黄酮在母丁香中尚未发现。更多还原

【Abstract】 Eugenia caryophylla Thunb., a commonly used herb, is documented in the Traditional Chineses Medicine (TCM) collection edition 2005. Flos Caryophyllate is the dry flower buds of Eugenia caryophylla Thunb., while Fructus Caryophylli is the mature fruits of Eugenia caryophylla Thunb.. It has been clearly known that effective components of these herbs are their secondary metabolites. The present paper comparatively analyzed chemical components of Flos Caryophyllata and Fructus Caryophylli by high-speed counter-current chromatography technique.This dissertation consists of three sections:The first part is the introduction section. In this section, an extensive review including chemical components, pharmaceutical effect, purification methods of Flos Caryophyllate and Fructus Caryophylli was included. A brief review about the latest progress of HSCCC was also included in this section.The second section focuses on Comparative analysis of Flos Caryophyllata and Fructus Caryophylli by high-speed counter-current chromatography. The herbs were first extracted by 95% ethanol continous reflux. The crude extracts were further extracted by petroleum ether and ethyl acetate sequentially, followed by macroporous resin fractionation. The fractions were then separated by HSCCC in a stepwise elution technology respectively. The two-phase solvent system composed of hexane: ethyl acetate: methanol: water (5:8:6:13, v/v/v/v, A solvent system) and hexane: ethyl acetate: methanol: water (5:8:9:10, v/v/v/v, B solvent system) that were optimized and determined according to their physical parameters. The separation was initially done by filling the column with upper phase of A solvent system as the stationary phase followed by elution with lower phase of A solvent system for 210 min. Afterwards it eluted with lower phase of B solvent system for another 340 min. Rotation speed of the chromatography was 880 r/min and the flow rate maintains 1.2 mL/min. The sample volume was 70 mg. Nine peaks were identified among which 6 peaks were consistent from both samples. The average of relative standard deviation (RSD) from the 6 peaks’HSCCC retention times was 2.260%, which is much lower than the national standared criteria 3%. Overlay ratios between two HSCCC chromatograms were around 66.67%, which manifest efficiency of HSCCC method in the test. In addition, five compounds including ellagic acid, 5,7-dimethoxy-2-methyl-chromen-4-one, 5,7-dimethoxy- 2,6-dimethyl- chromen -4-one, 3,5,3’4’-tetrahydroxy-7-methoxyflavone and quercetinpurified from the two herbs, among which quercetin, ellagic acid and 3,5,3’4’-tetrahydroxy-7-methoxyflavone are known compounds isolated from Eugenia caryophylla Thunb.. To be mentioned here, 5,7-dimethoxy- 2,6-dimethyl-chromen-4-one is a compound that firstly identified by NMR information so far.Based on the second section, compounds from Fructus Caryophylli was systematically isolated by using HPLC method. Three compounds including gallic acid, bancroftione and recrystallized 5,7,3’,4’,5’-pentamethoxyflavanone were purified. Among which 5,7,3’,4’,5’-pentamethoxyflavanone and was firstly identified from Fructus Caryophylli.更多还原