银杏叶中聚戊烯醇含量及其季节性变化

银杏 (ＧｉｎｋｇｏｂｉｌｏｂａＬ .)叶中聚戊烯醇 (ｐｏｌｙｐｒｅｎｏｌｓ)含量较高[1] ,聚戊烯醇在人体中是多萜醇的中间体[2 ] ,对细胞膜糖蛋白生物合成具有重要作用[3 ] ,用于多发性硬化症 (痛风 ,红斑狼疮等 )等免疫功能疾病、糖尿病、慢性肝炎及肿瘤病人化疗的辅助治疗     

Water-soluble polysaccharides from Ginkgo biloba leaves.

The water-soluble polysaccharides from dried Ginkgo biloba leaves were isolated after exhaustive extraction with organic solvents. The polysaccharide mixture could be separated into a neutral (GF1) and two acidic (GF2 and GF3) polysaccharide fractions by ion exchange chromatography. According to the Mr distribution GF1 and GF3 seemed to be homogenous, whereas GF2 could be further fractionated into two subfractions (GF2a and GF2b) by gel permeation chromatography. GF1 (Mr 23,000) showed the structural features of a branched arabinan. The main chain was composed of 1,5-linked arabinose residues and three in 12 arabinose molecules were branched via C-2 or C-3. GF2a (Mr 500,000) consisted mainly of 1,2,4-branched mannose (29%), 1,4-linked glucuronic (32%) and galacturonic (8%) acid as well as terminal rhamnose (25%). After removal of ca 70% of the terminal rhamnose the remaining polysaccharide showed a decrease in 1,2,4-branched mannose and an increase in 1,2-linked mannose indicating that at least half of the rhamnose residues were linked to mannose via C-4. GF3 (Mr 40,000) consisted of 1,4-linked galacturonic (30%) and glucuronic (16) acid, 1,3,6-branched galactose (15%), 1,2-linked (5%) and 1,2,4-branched (3.5%) rhamnose as well as 1,5-linked arabinose (11%). Rhamnose (5%) and arabinose (10%) were present as terminal groups. Mild acid hydrolysis selectively cleaved arabinose and the remaining polysaccharide showed an increased amount of 1,6-linked and terminal galactose and a decreased quantity of 1,3,6-branched galactose. These results indicated that the terminal as well as the 1,5-linked arabinose were mainly connected to galactose via C-3. The GF3 polysaccharide appeared to be a rhamnogalacturonan with arabinogalactan side chains.     

Coumaroyl flavonol glycosides from the leaves of Ginkgo biloba.

Two coumaroyl flavonol glycosides, isorhamnetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], and kaempferol 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside]-7-O-beta-D-glucopyranoside, were isolated from the n-BuOH extract of Ginkgo biloba leaves. These two, together with six other flavonol glycosides, kaempferol 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], quercetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], quercetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside]-7-O-beta-D-glucopyranoside, quercetin 3-O-beta-D-glucopyranosyl-(1-2)-alpha-L-rhamnopyranoside, quercetin 3-O-beta-rutinoside, and quercetin 3-O-beta-D-glucopyranoside, showed profound antioxidant activities in DPPH and cytochrome-c reduction assays using the HL-60 cell culture system.     

Long-chain polyprenols in gymnosperm plants

Over 100 species of gymnosperm plants were checked for the presence of long chain polyprenols. Poly-cis long chain prenols, mainly as acetates, were found in green needles of about 60 species. In Cycadopsida either prenol-18 or prenol-20 were the main components of the natural polyprenol mixture. In Coniferopsida either a single polyprenol family was present like in all species of Pinaceae, or polyprenols consisted of two families differing in the size of polyprenol molecules: one family in which prenol-17 was the dominating component, and the other family of prenol-23. These complex mixtures of polyprenols were present in Araucariaceae, Cupressaceae, Taxodiaceae and also in Taxopsida. Seasonal variations were observed in the polyprenol content in green leaves.     

银杏叶单组分双黄酮分离制备及抗氧化活性研究

从银杏叶中分离制备高纯度双黄酮对照品,探讨其抗氧化活性强弱。以60%乙醇-水提取的银杏叶浸膏为原料,反溶剂沉淀得到含量80.40%的双黄酮粗品,再以甲醇-水为流动相,半制备色谱梯度洗脱。结果得到了4种符合中药化学对照品要求的高纯度双黄酮,在最佳制备条件下,4种双黄酮纯度:阿曼托黄素(98.45%)、白果素(98.66%)、银杏黄素异构体(98.87%)、金松双黄酮(99.29%),其产率(mg/kg)分别为:22.5、23.8、192.5、71.8,考察了它们的抗氧化活性,并采用60%乙腈和有机酸水溶液对银杏黄素异构体进行了完全分离。建立的制备方法快速简便,所得单组分双黄酮纯度、产量高。本工作为单组分银杏双黄酮新药的研究与开发提供重要的技术支持。     

Ginkgolide B production in cultured cells derived from Ginkgo biloba L. leaves

Callus cultures and cell suspension cultures derived from Ginkgo biloba L. leaves produced ginkgolidc B. In cell suspension cultures, the production reached a maximum by the 13th day of subculture and followed by a sharp decrease. The medium of Murashige and Skoog induced the highest ginkgolide B content in cultures while the medium of Schenk and Hildebrandt promoted cell growth. For the maximal production of ginkgolide B, cells were cultured in Murashige and Skoog medium modified to contain 1.0 mg/l of -naphthaleneacetic acid, 0.1 mg/1 of kinetin, 30 g/1 sucrose and 1.25 mM potassium phosphate with a molar ratio of ammonium to nitrate ions of 1 3.Abbreviations B5 Gamborg et al (1968) medium - GKB Ginkgolide B - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic aicd - SH Schenk and Hildebrandt (1972) medium     

银杏叶黄酮提取方法比较

比较不同溶剂提取银杏（ＧｉｎｋｇｏｂｉｌｏｂａＬ．）叶黄酮类化合物的提取效率,从成本效益角度考虑,以７０％乙醇作为提取溶剂更为有利。在分级沉淀中,黄酮含量与蛋白质含量呈正相关,在乙醇提取液中黄酮和蛋白质含量最高;蛋白质的存在有助于提高黄酮的溶解度。乙醇提取液用饱和（ＮＨ４）２ＳＯ４浓缩两次,可使醇相中的黄酮沉淀析出。根据试验结果,提出了银杏叶黄酮的优化提取方法。     

Alkylresorcinols in fruit pulp and leaves of Ginkgo biloba L

These studies were undertaken to characterise resorcinolic lipids (5-n-alk(en)ylresorcinols) composition and to determine their seasonal fluctuations in fruit pulp and leaves of Ginkgo biloba L. Resorcinolic lipid concentrations were consistently higher in fruit pulp than in leaves. In pulp, several mono- and di-unsaturated homologs of alkylresorcinols were the predominant group of analysed lipids. Contrary to the fruit pulp, only 5-n-pentadecylresorcinol was demonstrated in leaves. Initially, the alkylresorcinol's content both in pulp and leaves increased until June-July and decreased following seeds ripening. This trend continued until senescence of leaves in late September and October.     

Deterrents extracted from the leaves of Ginkgo biloba: effects on feeding and contact chemoreceptors

Powdered dried ginkgo tree leaves were subjected to various chemical extractions and successive extracts were monitored for antifeedant activity against larvae of Pieris brassicae. Many fractions moderately inhibited food intake, and some were deterrent at levels as low as 25–50 ppm. Some behaviourally highly active fractions were tested electrophysiologically for neural responses in the maxillary taste sensilla. These extracts appeared to stimulate deterrent receptors. There were distinct differences in responses between Pieris brassicae and P. rapae. Ginkgolide A, B, and C each strongly stimulated deterrent receptors in P. rapae, which corresponds with the observation (Matsumoto & Sei, 1987) that these compounds are effective antifeedants for this species. No toxic effects were observed in insects after feeding for 24 h on diets containing ginkgo extracts.

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Résumé Des feuilles de Gingko séchées et broyées ont été soumises à différentes extractions. On a testé l'activité déterrante (antiapprétant) des extraits successifs sur les larves de Pieris brassicae. Un grand nombre de fractions inhibent modérément la consommation, et certaines sont déterrantes à des concentrations aussi faibles que 25–50 ppm. Par électrophysiologie, on a testé la réponse nerveuse des sensilles gustatives maxillaires à certaines fractions qui montraient une forte action sur le comportement. Il s'avère que ces extraits stimulent les récepteurs déterrants. Les réponses diffèrent pour P. brassicae et P. rapae. Les gingkolides A, B et C stimulent chacun fortement les récepteurs déterrants chez P. rapae, ce qui correspond aux observations (Matsumoto & Sei, 1987) selon lesquelles ces composés sont des antiappétants efficaces pour cette espèce. On n'a pas observé d'effet toxique sur les insectes nourris durant 24 heures par des diètes contenant des extraits de Gingko.

     

银杏叶多酚的机械力辅助提取及其体外抗氧化活性研究

为优化银杏叶多酚提取工艺,通过单因素试验考察填充率、球磨转速、球磨时间、乙醇浓度、料液比、提取温度、提取时间七个因素对机械力辅助提取银杏叶多酚得率的影响,以银杏叶多酚得率为响应值,采用Box-Benhnken三因素三水平响应面设计优化工艺,同时比较了4种提取方法对银杏叶多酚提取得率和抗氧化活性的差异。结果表明,机械力辅助提取银杏叶多酚的最佳工艺条件为:填充率26%、球磨转速为400rpm、球磨时间为15min。在此条件下,银杏叶多酚的得率为7.33%。机械力辅助乙醇提取银杏叶多酚得率低于碱水提取法,但是抗氧化活性高于碱水法提取的银杏叶多酚;抗氧化活性与乙醇回流法提取的银杏叶多酚相当,但是提取得率高于乙醇回流法。此提取工艺高效可行,具有一定的参考价值。     

Spatial and temporal variation of secondary metabolite profiles in Ginkgo biloba leaves

Terpene trilactones (TTLs) are the main bioactive constituents of Ginkgo biloba used for the preparation of drugs for several ailments. Flavonoid glycosides (FGs) are the significant group of compounds found in Ginkgo leaves used in food and healthcare products. The variation in the content of bioactive constituents, as well as antioxidant activity, with respect to change in altitude and the time of sample collection was investigated. The amount of TTLs varied from 0.218-0.709% w/w, whereas FGs were observed in the range of 0.130-0.209% w/w. Ginkgolides J and A showed significant variation (p ≤ 0.05) with the change in altitude, while other components remained more or less unaffected. The amount of TTLs and FGs were not affected significantly by the time of sample collection. The antioxidant capacity (by using DPPH, ABTS, and FRAP assays) of ginkgo extracts was highly dependent on the altitude, and was at maximum in the samples collected from 1000-2000 m, followed by the samples collected from altitudes lower than 1000 m and higher than 2000 m. Thus, Ginkgo harvested from 1000-2000 m of altitudinal range irrespective of the season could provide better nutraceutical formulations, whereas that from below 1000 m may yield a terpene trilactone-enriched fraction.     

Two new nonacosanetriols from Ginkgo biloba sarcotesta

Two new fatty alcohols named as (7S,8R,11S)-nonacosanetriol (1) and (10R,12R,15S)-nonacosanetriol (2), along with eight known compounds including ginkgolic acid (3), hydroginkgolic acid (4), sciadopitysin (5), ginkgetin (6), isoginkgetin (7), ginkgolide A (8), ginkgolide B (9) and ginkgolide C (10) have been isolated from the petroleum ether extract of Ginkgo biloba sarcotesta. Their structures were elucidated by means of chemical and extensive spectroscopic analysis. The absolute stereochemistry of compounds 1 and 2 was elucidated on the spectroscopic analysis of the R- and S-MTPA esters. Compounds 1 and 2 exhibited slight activity of antithrombin and moderate activity of antiplatelet aggregation in vitro. This was the first report regarding the anticoagulative activities of biflavonoids in G. biloba, and isoginkgetin (7) showed significant antithrombin and antiplatelet aggregation activity.     

Inhibition of fatty acid synthase by ginkgolic acids from the leaves of Ginkgo biloba and their cytotoxic activity

Fatty acid synthase (FAS) has been proposed to be a new drug target for the development of anticancer agents because of the significant difference in expression of FAS between normal and tumour cells. Since a n-hexane-soluble extract from Ginkgo biloba was demonstrated to inhibit FAS activity in our preliminary test, we isolated active compounds from the n-hexane-soluble extract and evaluated their cytotoxic activity in human cancer cells. Three ginkgolic acids 1–3 isolated from the n-hexane-soluble extract inhibited the enzyme with IC₅₀ values 17.1, 9.2 and 10.5 µM, respectively, and they showed cytotoxic activity against MCF-7 (human breast adenocarcinoma), A549 (human lung adenocarcinoma) and HL-60 (human leukaemia) cells. Our findings suggest that alkylphenol derivatives might be a new type of FAS inhibitor with cytotoxic activity.     

银杏叶超临界流体提取物对杉木种子的发芽效应

采用正交试验设计L8(2^7)及超临界萃取技术,乙醇作夹带剂,就银杏(Ginkgo biloba L.)叶提取物对杉木[Cunninghamia lanceolata (Lamb.) Hook.]种子的发芽效应进行了生物检测。结果表明：不同萃取条件下,纯CO2及夹带剂和CO2混合萃取的银杏叶提取物对杉木种子发芽的影响不同;就尉值(敏感指数)而言,在纯CO2萃取的浓度为200mg／kg银杏叶提取物中,杉木种子绝对发芽率为0．23,绝对发芽势为0．15,胚根长为0．01,胚轴长为-0．02;夹带剂和CO2混合萃取的浓度为200mg／kg银杏叶提取物除了对杉木种子绝对发芽率和绝对发芽势表现为促进作用外(尉值分别为0．03和0．34),对胚根长及胚轴长则表现为轻微的抑制作用(尉值分别为-0．09和-0．03)。     

银杏叶多糖研究进展

银杏叶多糖具有多种重要的生物活性和功能,是目前药物开发和研究的热点之一。对银杏叶多糖提取、分离纯化、结构分析和生物活性的研究现状进行综述,并指出研究存在的问题和今后的研究方向。     

Origin of Ginkgo biloba L. Phylogenetic approach

A cladistic analysis on fossil and modern Gymnosperms (20 taxa) is presented and discussed with particular mention of Ginkgo biloba L. origin. The consensus tree obtained from 68 characters (59 informative characters) shows a monophyletic clade containing all plants bearing micropylate ovules ('Micropylophytes'). Medullosales appear at the base of this clade. Ginkgo forms the sister group of the Dicranophyllales + Coniferales. The obtained phylogeny implies that the Ginkgoales ancestor is to be found during the Upper Carboniferous.     

Novel plastocyanin containing phenylalanine-83 from the gymnosperm Ginkgo biloba

Plastocyanin was purified from the gymnosperm Ginkgo biloba L., and its complete amino acid sequence was determined. The protein was shown to contain Phe-83 instead of Tyr-83 conserved in other land plant plastocyanins. This is the first report of the characterization and complete amino acid sequence of a gymnosperm plastocyanin.