Investigation of biologically active components in ginkgo leaf products on the Japanese market

The rapid and highly separative ultra high-performance liquid chromatography (UHPLC)-UV method was adopted and validated to investigate the flavonol glycoside compositions in ginkgo leaf products on the Japanese market. The result indicates that certain products contained amounts of flavonol glycosides approximately equivalent to the medicinal product. Additionally, we examined the correlations between the total amount of flavonol glycosides and of terpene lactones in various ginkgo leaf products.     

Isolation and antioxidant activity of galloyl flavonol glycosides from the seashore plant, Pemphis acidula

Four kinds of galloyl flavonol glycosides were found in the leaf extract of Pemphis acidula, a plant growing on the subtropical seashore. Their chemical structures were elucidated to be quercetin or kaempferol 6"-O-galloyl-beta-D-glycosides by using spectroscopic and chemical analyses. One of the flavonols, kaempferol-3-O-(6-O-galloyl-beta-D-galactopyranoside), was newly isolated from natural sources and its structure was completely determined in this investigation. The antioxidant-related activities of the galloyl flavonoids were examined by the DPPH antiradical activity, inhibition of methyl linoleate oxidation, and inhibition of oxidative cell death. These results were compared with those of the corresponding non-galloylated flavonol glycosides and their aglycones. The galloyl flavonoids showed more efficient activity than that of the corresponding flavonol glycosides, but not more than that of the corresponding aglycones in the three assays applied.     

Internal and external leaf flavonoids of Pericome caudata

Pericome caudata accumulates four flavonol aglycones externally on leaf and stem surfaces. The glycosides present in the leaf tissue are based on eight further flavonols. This preponderance of tissue flavonoids over exudate flavonoids is unusual.     

Identification of Terpene Lactones and Flavonol Glycosides in Preparations Based on Ginkgo Biloba Extract and a New Way of Semi-Quantitative Determination of Flavonol Glycosides by <Superscript>1</Superscript>H NMR Spectroscopy

The composition of terpen lactones and flavonol glycosides of commercial preparation series based on Ginkgo biloba extracts was investigated by ¹H NMR spectroscopy. The content of individual terpen lactones was determined using DMSO-d₆ and acetone-d₆ solvents. The effect of the structure of flavonol glycosides on the signal of the hydroxyl proton at a position 5 of the ring A was examined. A new approach was proposed for semiquantitative determination of the total amount of flavonol glycosides by the integral intensity of this signal, which is a superposition of the singlets in the region of 12.5–12.65 ppm of individual flavonoids in DMSO-d₆. Since the corresponding signals of aglycones (quercetin, kaempferol, isorhamnetin), which are minor components of the Ginkgo biloba extracts, appear separately in a slightly different region (12.45–12.48 ppm), the proposed method can also be used for detecting adulteration of Ginkgo biloba extracts by means of the addition into them of relatively cheap aglycones or rutin as well as for assessment of the content of flavonoids of similar structure in some types of plant raw materials.     

6-Methoxyflavonols from disjunct populations of Brickellia cylindracea (compositae)

Eight identical 6-methylated flavonols including aglycones and glycosides were isolated from two geographically disjunction population of Brickellia cylindracea from Mount Livermore and Austin, Texas, suggesting that they are best treated as a single taxon. Among the flavonol aglycones identified were patuletin, centaureidin, quercetagetin 3,6,3′,4′-tetramethyl ether and artemetin. The flavonol glycosides were patuletin and its 3-galactoside, 3-galactogalactoside, 3-rhamnogalactoside and a 3-rhamnogalactoside derivative.     

高效液相色谱法测定银杏叶提取物中黄酮甙含量

用高效液相色谱法测定银杏叶提取物经酸水解后黄酮甙元含量．采用Ｃ１８柱,甲醇：水：磷酸（５５：４４．５：０．５）为流动相,检测波长３７０ｎｍ,方法回收率９５．１％～１０３．２％,变异系数（ＣＶ）３．１７％,１０批样品含测平均为２５．４％．     

A novel substance with allelopathic activity in Ginkgo biloba

Ginkgo (Ginkgo biloba) is one of the oldest living tree species and has been widely used in traditional medicine. Leaf extracts of ginkgo, such as the standardized extract EGb761, have become one of the best-selling herbal products. However, no bioactive compound directed at plants has been reported in this species. Therefore, we investigated possible allelopathic activity and searched for allelopathically active substances in ginkgo leaves. An aqueous methanol leaf extract inhibited the growth of roots and shoots of garden cress (Lepidium sativum), lettuce (Lactuca sativa), timothy (Phleum pratense) and ryegrass (Lolium multiflorum) seedlings. The extract was purified by several chromatographic runs and an allelopathically active substance was isolated and identified by spectral analysis to be the novel compound 2-hydroxy-6-(10-hydroxypentadec-11-enyl)benzoic acid. The compound inhibited root and shoot growth of garden cress and timothy at concentrations greater than 3 μM. The activity of the compound was 10- to 52-fold that of nonanoic acid. These results suggest that 2-hydroxy-6-(10-hydroxypentadec-11-enyl)benzoic acid may contribute to the allelopathic effect caused by ginkgo leaf extract. The compound may also have potential as a template for the development of new plant control substances.     

Chemosystematic studies on certain species of the family Brassicaceae (Cruciferae) in Egypt

The flavonoids of nine selected species belonging to different tribes of family Brassicaceae (Cruciferae) native to Egypt were surveyed, viz. Rorippa palustris, Coronopus squamatus, Eremobium aegyptiacum, Moricandia nitens, Brassica tournefortii, Farsetia aegyptia, Matthiola livida, Anastatica hierochuntica and Sisymbrium irio. Thirty-eight compounds were isolated and identified, which included six flavonol aglycones, 24 flavonol glycosides including 14 flavonol 3,7-diglycosides, one flavone aglycone, three flavone O–glycosides, two glycoflavones and two dihydroflavonoids. A numerical analysis based on a combination of 97 morphological, anatomical and chemical characters revealed two series, two subseries, two clusters and two groups. The interrelationships between the studied species are discussed.     

Flavonoid diversity and endemism in Arnica subgenus Austromontana

Eighty-seven populations, representing eight species of Amica subgenus Austromontana, were examined for flavonoids. A total of 22 compounds, including simple and methylated flavone and flavonol glycosides as well as methylated aglycones, were isolated and identified. The two widespread species, A. cordifolia and its presumed early derivative A. latifolia, are ancestral species of the subgenus. From these two species, major evolutionary diversification within the subgenus has taken place largely in the Klamath region of Oregon and California, resulting in the formation of several rare, endemic species. These endemics are generally characterized by reduced flavonoid profiles and/or an increase in the number of methylated aglycones while, in contrast, the wider ranging species have few methylated aglycones and more glycosides. However, one species, A. viscosa, a rare volcanic endemic, exhibits the most elaborate and specialized flavonoid profile within the subgenus.     

Purification and identification of flavonoids from the yellow green cocoon shell (Sasamayu) of the silkworm,Bombyx mori

Three quercetin glycosides, quercetin 5-O-beta-D-glucoside, quercetin 7-O-beta-D-glucoside, and quercetin 4'-O-beta-D-glucoside, and two kaempferol glycosides, kaempferol 5-O-beta-D-glucoside and kaempferol 7-O-beta-D-glucoside, along with their aglycones, quercetin and kaempferol, were isolated from an ethanolic extract of Sasamayu cocoon shells. The chemical structures were characterized by chemical and spectroscopic methods including UV spectrometry and HPLC-ESI-MS. The five flavonol glycosides of the shell are different structurally from those of the leaves of mulberry (Morus alba). It was suggested that potent antioxidative activity in the cocoon is mainly due to flavonoid compounds since free radical scavenging activity was found in the cocoon flavonoids identified here.     

Leaf flavonoids of Amborella trichopoda

The leaf flavonoids of Amborella trichopoda were examined and two kaempterol glycosides were detected. Procyanidin was also present. These results are similar to the flavonoid pattern in other families of the Laurales and it is suggested that simple flavonol glycosides are a primitive feature in the order.     

Flavonol glycosides of Limnanthes douglasii

Eighteen flavonol glycosides were isolated from petal and leaf-stem of Limnanthes douglasii. There were six aglycones: kaempferol, quercetin, isorhamnetin, myriectin, syringetin and a new flavonol, myricetin 3′-methyl ether. Each occurred as the 3-rutinoside, 3-rhamnosylrutinoside and 3-rutinoside-7-glucoside.     

Purification and Identification of Flavonoids from the Yellow Green Cocoon Shell (Sasamayu) of the Silkworm,Bombyx mori

Three quercetin glycosides, quercetin 5-O-β-D-glucoside, quercetin 7-O-β-D-glucoside, and quercetin 4′-O-β-D-glucoside, and two kaempferol glycosides, kaempferol 5-O-β-D-glucoside and kaempferol 7-O-β-D-glucoside, along with their aglycones, quercetin and kaempferol, were isolated from an ethanolic extract of Sasamayu cocoon shells. The chemical structures were characterized by chemical and spectroscopic methods including UV spectrometry and HPLC-ESI-MS. The five flavonol glycosides of the shell are different structurally from those of the leaves of mulberry (Morus alba). It was suggested that potent antioxidative activity in the cocoon is mainly due to flavonoid compounds since free radical scavenging activity was found in the cocoon flavonoids identified here.     

贵州传统发酵豆制品中水解银杏黄酮苷的微生物β-葡萄糖苷酶筛选

[目的]微生物β-葡萄糖苷酶法水解银杏黄酮苷具有重要意义,不过目前这方面的研究极少。因此,本文目的是筛选到水解银杏黄酮苷的酶活高的微生物β-葡萄糖苷酶,并分析其底物选择性机制。[方法]以银杏叶提取物作为唯一碳源富集培养,从贵州传统发酵豆豉中筛选产对银杏黄酮苷水解酶活高的β-葡萄糖苷酶的菌株,并对该菌株进行鉴定。然后比较此β-葡萄糖苷酶对不同底物的选择性,同时测定此酶水解银杏黄酮苷反应的米氏常数Km及最大反应速率Vmax。最后,对不同的底物进行分子对接,分析其底物特异性机制。[结果]结果表明,筛选到的菌株GUXN01所产β-葡萄糖苷酶水解银杏黄酮苷的酶活最高,被鉴定为枯草芽孢杆菌。此β-葡糖糖苷酶对β构型的糖类以及苷类等具有广泛的底物特异性和不同的选择性,尤其对银杏黄酮苷具有很好的亲和性。分子对接研究表明枯草芽孢杆菌β-葡萄糖苷酶对银杏黄酮苷和其他糖苷类具有不同亲和性和选择性的原因主要是酶结构和底物分子结构的相互作用力的差异导致的。[结论]这些发现为GUXN01所产的β-葡萄糖苷酶应用于水解银杏黄酮苷类生产相应苷元奠定了良好的基础。     

Phenolic composition and content of representatives of genus Spiraea L. under industrial pollution in Novosibirsk

The composition and content of groups of phenolic compounds (free flavonol aglycones, flavonol glycosides, free and bound phenolic acids, hydroxybenzoic acids, hydroxycinnamic acids, and their esters) and individual components (quercetin; kaempferol; rutin; hyperoside; isoquercitrin; avicularin; and gallic, protocatechuic, vanillic, salicylic, chlorogenic, p-coumaric, o-coumaric and cinnamic acids) in leaves of Spiraea media Fr. Schmidt, S. chamaedryfolia L., and S. hypericifolia L. growing in areas of Novosibirsk with high and background levels of industry pollution have been studied. Differences in phenolic composition and content are revealed. The total phenolic content in leaves of the studied species in pollution is 1.6 (S. hypericifolia) and 2.4 (S. media and S. chamaedryfolia) fold lower than the background conditions. The decrease in the phenolic content in S. media are due to flavonol glycosides (mostly rutin), and in S. chamaedryfolia and S. hypericifolia it is due to hydroxycinnamic acids and their esters.     

Profiling changes in metabolism of isoflavonoids and their conjugates in Lupinus albus treated with biotic elicitor

Liquid chromatography with ultraviolet and mass spectrometric detection was applied to monitor changes in profiles of isoflavonoid glycosides and free isoflavonoid aglycones in Lupinus albus L. Four isoflavonoid aglycones, fourteen isoflavonoid glycosides, four flavonol glycosides and flavone glycoside were identified in lupin tissue after LC/ESI/MS analyses. An elicitor preparation from purified yeast cell wall was used to inject the shoots of 3-week old seedlings or to infiltrate the cut lupin leaves. Qualitative and quantitative changes of isoflavonoids were measured at different time points after elicitation. In elicited lupin seedlings increased amounts of prenylated isoflavone aglycones were identified. The concentrations of glycosidic conjugates of isoflavones present in plant tissue were less affected.     

Systematic implications of flavonoids in Hazardia

The flavonoid patterns in Hazardia species support species delimitations and relationships based on morphology and geography. The compounds thus far elucidated are glycosides of quercetin, kaempferol, isorhamnetin, luteolin, and apigenin, glycoflavones of apigenin, and methoxylated flavonol aglycones.     

Evolution des flavonoides au cours de la croissance des bractees de Poinsettia

The changes in flavonoids were studies during the growth of Poinsettia bracts. Pelargonidin glycosides appeared after cyanidin glycosides; the 3-rutinoside after the 3-monoglucoside. The 3-mono-glucosides were predominant at all times. The bracts contained kaempferol and quercetin, both as aglycones and glycosides. There was no direct relationship between the pathways of flavonol and anthocyanin biosynthesis, but two separate pathways corresponding to the 4′-monohydroxylated- and 3′,4′-dihydroxylated-, flavonoids. Flavanonols and isoflavones were detected but not characterized.     

Five Nodulation Mutants of White Sweetclover (Melilotus alba Desr.) Exhibit Distinct Phenotypes Blocked at Root Hair Curling,Infection Thread Development,and Nodule Organogenesis

Flavonol aglycones are required for pollen germination in petunia (Petunia hybrida L.). Mutant plants lacking chalcone synthase (CHS), which catalyzes the first committed step in flavonoid synthesis, do not accumulate flavonols and are self-sterile. The mutant pollen can be induced to germinate by supplementing it with kaempferol, a flavonol aglycone, either at the time of pollination or by addition to an in vitro germination system. Biochemical complementation occurs naturally when the mutant, flavonol-deficient pollen is crossed to wild-type, flavonoid-producing stigmas. We found that successful pollination depends on stigma maturity, indicating that flavonol aglycone accumulation may be developmentally regulated. Quantitative immunoblotting, in vitro and in vivo pollen germination, and high-performance liquid chromatographic analyses of stigma and anther extracts were used to determine the relationship between CHS levels and flavonol aglycone accumulation in developing petunia flowers. Although substantial levels of CHS were measured, we detected no flavonol aglycones in wild-type stigma or anther extracts. Instead, the occurrence of a conjugated form (flavonol glycoside) suggests that a mechanism may operate to convert glycosides to the active aglycone form.