摘要
研究课题一基于生物活性菧类的树豆叶质量标准研究
研究背景:树豆(Cajanus cajan (L.) Millsp.)为蝶形花科(Papilionaceae)木豆属(Cajanus)植物,主要分布于亚洲、非洲和美洲的热带和亚热带地区。我国树豆资源丰富,主要分布于云南、海南、广西、贵州、广东、福建、湖南、四川、江西、浙江、台湾等省,尤以云南省为多。树豆主要有清热解毒、消炎抗菌、止血解痛、强骨等功效,可用于治疗外伤、烧伤、感染、糖尿病、骨质疏松、肿瘤、肝炎、麻疹、疟疾和痢疾等症。国内外学者对树豆的活性成分进行了一系列的研究表明,其主要化学成分为黄酮类(flavonoids)及菧类化合物(stilbenes)。这两类化合物被认为是树豆的主要活性成分。
目的:菧类化合物Longistylin A和longistylin C是仅存在于Lonchocarpus longistylus和树豆等少数几种豆科植物的特征化学成分和药理活性指标成分。本实验首次对不同产地来源的树豆叶中longistylin A和longistylin C进行定性检测与含量测定。方法:以二氯甲烷为硅胶薄层色谱展开剂,以3%香草醛-6%高氯酸溶液为显色剂,可以清晰检测指标成分;采用Thermo BDS Hypersil C18色谱柱(4.6mm×250mm,5μm),以甲醇-水(80:20)为流动相可使指标成分获得良好分离;结果:longistylin A和longistylin C分别在0.00288-0.0576μg和0.0112-0.224μg范围呈良好的线性关系,平均回收率分别为98.9%,97.2%,RSD分别为2.43%,2.23%。结论:建立了以树豆叶中药物活性菧类成分为指标的定性定量分析方法,可用于树豆(木豆)药材的质量控制。
研究课题二黄花夹竹桃强心苷的降解产物
研究背景和目的:黄花夹竹桃(Thevetia peruviana)是央竹桃科(Apocynaceae)黄花夹竹桃属(Thevetia Linn.)。分布于热带美洲和热带亚洲,现全世界热带和亚热带地区均有栽培,我国引入栽培的有黄花夹竹桃Thevetia. peruviana (Pers.) K. Schum.和Thevetia.ahouai (L.) A. DC2种。主要在广东、广西、福建、台湾、云南等南方各省广为栽培,长江流域及其以北行温室栽培。自1881年De Vry从黄花夹竹桃中提出-种水溶性苷,命名为thevetin以来,关于黄花夹竹桃成分的研究取得了重要进展,从黄花夹竹桃种子,茎叶和根中获得了多种化合物,主要有强心苷、脂肪油,糖类,黄酮类及环烯醚萜类等,其中强心苷具有多种生物活性,最新研究进展表明黄花夹竹桃降解所得次苷有体外抗肿瘤作用,为了进一步寻找该植物的有效成分,本研究课题对黄花央竹桃用不同方法进行降解,并对降解产物进行了成分分离、鉴定和体外抗肿瘤作用研究,以期有新的发现,为黄花夹竹桃的深入研究和开发应用奠定基础。
方法:本实验利用生物发酵法、液液萃取、正相硅胶柱、反相ODS柱等柱层析手段,结果:从黄花夹竹桃的二氯甲烷萃取部位分离强心甾内酯化合物,并采用1HNMR、13C NMR等波谱学测试方法,结合ESI-MS质谱和元素分析,鉴定其化学结构。结论:从黄花夹竹桃中分离得到4个强心甾内酯化合物Cerberin、Neriifolin、Peruvoside、 Thevebioside。
Part One:Studies on Quality Standard of the leaves of Cajanus cajan based on bioactive Stilbenes
Background:Cajanus cajan (L.) Millsp belongs to a plant of Cajanus (Papilionac eae). It is mainly distributed in tropical and subtropical regions of Asia, Africa and the Americas. In china Cajanus cajan is an agricultural resource crop planted in Yu nnan, Hainan, Guangxi, Guizhou, Guangdong, Fujian, Hunan, Sichuan, Jiangxi, Zheji ang, Taiwan and other provinces. Its leaves are also used as materia medica in use of anti-inflammatory, anti-bacterial, strong bones, and treating trauma, burns, infectio n, diabetes, osteoporosis, cancer, hepatitis, measles, malaria and dysentery embolism. According to recent research flavonoids and stilbenes are considered to be main bi oactive ingredients of the leaves of Cajanus cajan.
Objective:The Longistylin A and longistylin C, two stilbene compounds isolated from the leaves of Cajanus cajan, are characteristics chemical composition exist onl y in the Lonchocarpus longistylus, Cajanus cajan, and other few legumes. In this st udy, we established a quality control criteria for medicinal herb Cajanus cajan base d on the qualitative and quantitative determination of longistylin A and longistylin C,
Method:They were detected by silica gel TLC with dichloromethane as developin g solvent, a3%vanillin-6%HC104in50%ethanol solution as visualization reagent.
Result:Longistylin A and Longistylin C exhibited good resolution on a Thermo BD S Hypersil C18column (4.6mm×250mm,5μm) with a mobile phase methanol-water (8:2), and showed good linearity in the range of0.00288~0.0576μg and0.0112~0.224μg respectively. The average recovery was98.9%or97.2%with RSD of2.43%or2.23%for longistylin A or longistylin C respectively. Conclusion:The esta blished qualitative and quantitative analyses are simple and accurate, and provide a base for quality control of Cajanus cajan (L.) Millsp.
Part Two:Studies on the Degraded Cardenolides from Thevetia peruviana
Background and Objective:Yellow oleander(Thevetia peruviana) belongs to Genus Thevetia of Apocynaceae Family. It is mainly distributed in tropical America and tropical Asia and cultivated in tropical and subtropical regions all around the world.Two kinds of species including Thevetia peruviana (Pers.) K. Schum and Thevetia.ahouai (L.) A. DC was introduced in China for cultivation, Mainly in the southern provinces of Guangdong, Guangxi, Fujian, Taiwan, Yunnan, and greenhouse cultivation in the north of Yangtze river and its watershed. Since1881, a water-soluble glycosides, named thevetin was isolated from the yellow oleander by De Vry, the research on the constituent of yellow oleander have made important progress. Variety of compounds including cardiac glycosides, fatty oils, sugars, flavonoids, and iridoid terpenes was obtained from yellow oleander seeds, stems, leaves and root, in which cardiac glycosides with strong biological activity. To further find the active ingredient in Thevetia peruviana, In this paper, degradation,isolation, purification, structural elucidation, in vitro activities of the degraded constituents from Thevetia peruviana against tumor cells were investigated to lay the foundation for in-depth research and development applications of the yellow oleander.
Method:In this study, cardiac steroidal lactone compounds from the dichloromethane extract parts of the yellow oleander were isolated by the means of bio-fermentation, the liquid-liquid extraction, normal phase silica gel and reversed phase ODS column chromatography, Result and Conclusion:From the physico-chemical data, including1HNMR,13CNMR ESI-MS, UV and IR, the chemical structures of the compounds were determined as Cerberin、Neriifolin、Peruvoside、Thevebioside.
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