摘要
葛枣猕猴桃[Actinidia Polygama (Sieb. et Zucc.) Miq]系猕猴桃科(Actinidiaceae)猕猴桃属(Actinidia)落叶缠绕藤木,分布于我国东北、华北,朝鲜、日本及苏联远东地区。葛枣猕猴桃果实入药,秋季采收,晒干即可。具有理气止痛之功能,主治腰痛、疝痛。葛枣猕猴桃枝叶辛温有小毒,可治大风癞疾,症积,气痢,风劳。
本实验对葛枣猕猴桃叶的化学成分进行了系统研究,利用硅胶、聚酰胺以及ODS柱层析、重结晶等手段从葛枣猕猴桃叶的乙醇提取物中分离得到了8个化合物,经理化性质和波谱数据分析,鉴定这些化合物分别为:山奈酚(Kaempferol)(Ⅰ),胡萝卜苷(Ⅱ),伞形花内酯(umbelliferone)(Ⅲ) ,山奈酚-3-O-β-D-吡喃半乳糖苷(kaempfero-l-3-O-β-D-galactopyranoside)(Ⅳ),异鼠李素-3-O-β-D-吡喃葡萄糖苷(isorhamnetin-3-O-β-D-glucoside)(Ⅴ),山奈酚-3-O-α-L-鼠李糖基-(1→6)-β-D-吡喃半乳糖苷( kaempferol-3-O-α-L-rhamnopyranosy-(1→6)-β-D-galactopyranoside )(Ⅵ) ,芹菜素-6-C-β-D-吡喃葡萄糖基-8-C-吡喃木糖苷(6-C-glycopyranosyl-8-C-xylopyranosyl apigenin)(Ⅶ),山奈酚-3-O-α-L-鼠李糖基- (1→3) -α-L-鼠李糖基- (1→6) -β-D-吡喃半乳糖苷(kaempferol-3-O-α-L-rhamnopyranosyl- (1→3) -α-L-rhamnopyranosyl - (1→6) -β-D - galactopyranoside) (Ⅷ)。化合物Ⅰ-Ⅷ均是首次从该植物中分离得到。
本实验还建立了葛枣猕猴桃叶中山奈酚-3-O-α-L-鼠李糖基-(1→3) -α-L-鼠李糖基-(1→6)-β-D-吡喃半乳糖苷(化合物Ⅷ)含量的HPLC测定方法,为建立葛枣猕猴桃叶质量标准及葛枣猕猴桃的开发利用提供了新的科学依据。
Actinidia Polygama (Sieb. et Zucc.) Miq pertain to the family of Actinidiaceae and genus of Actinidiaceae. Being a familiar fruit it also has important nutritive and medicinal value. For the purpose of improving its Quality Standard, in this paper the extraction, separation and structure identification for the chemical compositions of the leaves were carried out, and the quantitative determination of main chemical components was porformed as well. Eight compounds were obtained, they are identified as Kaempferol (I) ,daucossterol (II) , umbelliferone(III) , kaempfero-l-3-O-β-D- galactopyranoside (IV), isorhamneti -3-O-β-D-glucoside(V) , kaempferol- 3-O-α-L-rhamnopyranosy- (1→6)-β-D-galactopyranoside ( VI ), 6-C- glycopyranosyl-8-C- xylopyranosyl apigenin(Ⅶ),kaempferol-3-O-α-L - rhamnopyranosyl - (1→3) -α- L - rhamnopyranosyl - (1→6) -β-D - galactopyranoside(Ⅷ),respectively.
Besides, Kaempferol - 3-O-α-L- rhamnopyranosyl- (1→3) -α-L - rhamnopyranosyl - (1→6) -β-D-galactopyranoside(compound VIII)as a major compound was used for content determination in this paper. The determination method was proved to be an measurable method with the characters of accuracy, sensitivity and selectivity.
Extraction, Separation and Structure Identification
Dried hurb (1.0 kg) were extracted by heat reflux in 85% alcohol for three times, combined and concentrated to cream (100 g). The cream was subjected to silica gel column. Eight compounds[Ⅰ( 30mg),Ⅱ(50mg),Ⅲ(50mg),Ⅳ(60mg),Ⅴ(50mg),Ⅵ(28mg),Ⅶ( 123mg),Ⅷ(700mg)] were obtained after purified by silicagel,polyamide and ODS column chromatography repeatly.
CompoundsⅠ,Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ,ⅦandⅧwere identified respectively by UV, IR, MS, 1DNMR and 2DNMR as Kaempferol (Ⅰ) , daucossterol (Ⅱ) , umbelliferone (Ⅲ), kaempfero-l-3-O-β-D- galactopyranoside(Ⅵ) , isorhamnetin-3-O-β-D-glucoside(Ⅴ),kaempferol-3-O-α-L-rhamnopyranosy- (1→6) -β-D-galactopyranoside(Ⅵ),6-C-glycopyranosyl -8-C- xylopyranosyl apigenin(Ⅶ),kaempferol -3 -O -α-L - rhamnopyranosyl - (1→3) -α-L- rhamnopyranosyl - (1→6) -β-D -galactopyranoside(Ⅷ). Content determination of kaempferol -3 -O-α-L- rhamnopyranosyl - (1→3) -α- L- rhamnopyranosyl - (1→6) -β-D-galactopyranoside(compound VIII) in the Actinidia Polygama (Sieb. et Zucc.) Miq
In this paper we present a more sensitive quantification method by RP-HPLC for determining the compound VIII in the leaves of Actinidia Polygama (Sieb. et Zucc.) Miq.
HPLC was performed on a ZORBAX Eclipse XDB-C 18 column(250 mm×4.6 mm,5μm)at 25℃with the rate of 1.2ml/min,using 0.1% methanol solution as mobile phase (gradient elution: 0～30 min 37:63,30～50 min methanol).
ⅰThe linear range of compound VIII is 0.099～9.99μg. Y = 981.70 X + 11.37 r=0.99992
ⅱCompound VIII solution was injected into HPLC for six times, the RSD of precision tests is 0.30 % ( n=6).
ⅲThe repeatability tests for six samples showed RSD is 1.97% ( n=6).
ⅳThe test of stability showed its RSD is 0.53%, so the compound VIII is stable in sample solution during 12 hours.
ⅴThe average recovery is 102.3%, its RSD is 1.5 %( n=6).
ⅵThe contents of three batches of samples are 8.35,7.98,8.51mg/g respectively. Compound VIII are 1.65%,1.57%,1.49% respectively
In conclusion, all the results of this paper provided the foundations of quality standard establishment ,development and utilization of Actinidia Polygama (Sieb. et Zucc.) Miq.
引文
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