Characterization of flavonoids in the traditional Chinese herbal medicine-Huangqin by liquid chromatography coupled with electrospray ionization mass spectrometry

The root of Scutellaria baicalensis, called Huangqin in Chinese, is one of the most commonly used traditional Chinese medicines for the treatment of hepatitis, tumors, diarrhea, and inflammatory diseases. The major chemical constituents of Huangqin are flavonoids. In the present paper, HPLC-DAD-ESI-MS(n) was used to analyze flavonoids in the roots of S. baicalensis. A total of 26 flavonoids were identified or tentatively characterized, including 5 C-glycosides, 12 O-glycosides, and 9 free aglycones. Two C-glycosides, apigenin-6-C-glucyl-8-C-arabinoside and chrysin-6,8-di-C-glucoside, together with some O-glycosides, are reported from S. baicalensis for the first time. This method is simple, reliable and sensitive, and could be used for the quality control of Huangqin and its related preparations.     

Ethanolic crude extract and flavonoids isolated from Alternanthera maritima: neutrophil chemiluminescence inhibition and free radical scavenging activity

Extracts from Alternanthera maritima are used in Brazilian folk medicine for the treatment of infectious and inflammatory diseases. Bioassay-guided fractionation of A. maritima aerial parts yielded an ethanolic crude extract, its butanolic fraction and seven isolated flavonoids (two aglycones, two O-glycosides and three C-glycosides) with antioxidative activity. The ability of these samples to scavenge enzymatically generated free radicals (luminol-horseradish peroxidase-H2O2 reaction) and inhibit reactive oxygen species (ROS) production by opsonized zymosan-stimulated human neutrophils (PMNLs) was evaluated by chemiluminescence methods. In both assays, the butanolic fraction was significantly more active than the ethanolic crude extract, the flavonoid aglycones had high inhibitory activities and the C-glycosylated flavonoids had no significant effect even at the highest concentration tested (50 micromol/L). However, the O-glycosylated flavonoids inhibitory effects on chemiluminescence were strongly dependent on the chemical structure and assay type (cellular or cell-free system). Under the conditions tested, active samples were not toxic to human PMNLs.     

莲子心碳苷黄酮与黄酮异构体的色谱质谱研究

采用电喷雾质谱法(ESI-MS),对从莲子心分离得到的碳苷类黄酮化合物进行质谱碎裂规律研究。结果表明,负离子模式下,六碳糖碳苷黄酮主要发生糖环裂解,通过丢失特征性的碎片(90 u、120 u、150 u)与氧苷黄酮区分;单糖取代的六碳醛糖氧苷黄酮直接丢失单糖部分(162 u),六碳醛糖种类无法通过质谱区分,但由于它们在液相上的保留时间不同,可通过液相色谱-质谱(LC-MS)联用方法分离鉴定;二糖取代的氧苷黄酮主要碎片离子通过丢失糖部分(146 u、162 u、308 u)所得,二糖的种类及连接方式可通过质谱图上的碎片离子峰及其相对丰度辨别。莲子心中多种碳苷黄酮和氧苷黄酮质谱的不同裂解规律,不仅有助于莲子心黄酮化合物的快速鉴定,而且可以通过与液相色谱联用实现莲子心中同分异构体的快速区分。     

Epicuticular flavonoids of some Scrophulariaceae

Twenty-two species of Scophulariaceae have been found to accumulate flavonoid aglycones externally on their leaves and stems. They belong to the genera Anarrhinum, Antirrhinum, Asarina, Calceolaria, Mimulus, and Odontites. Most of the flavonoids are methylated flavones and flavonols, some with 6-O and/or 8-O-substitution. One of them is the natural isobutyryl ester of a rare flavone.     

Chemosystematic studies of Nitraria retusa and selected taxa of Zygophyllaceae in Egypt

Nitraria retusa (Forssk.) Asch. was subjected to botanical and chemical studies and compared against four selected taxa of Zygophyllaceae viz., Fagonia arabica L., Peganum harmala L., Tribulus terrestris L. and Zygophyllum album L.f. The botanical section included the study of morphological and anatomical characters of both vegetative and reproductive organs using LM and scanning electron microscope. The chemical section included a detailed study of the flavonoids present in N. retusa. Twelve compounds were isolated and identified as flavone, flavonol O-glycosides and flavone C-glycosides. Flavonoid patterns from N. retusa (Forssk.) Asch. of the selected taxa Zygophyllaceae were surveyed through HPLC and correlated with morphological and anatomical criteria.     

Analysis of flavonoids in pubescence of soybean near-isogenic lines for pubescence color loci

T and Td loci control pubescence color of soybean with epistatic effects (TT TdTd, tawny; TT tdtd, light tawny or near-gray; tt TdTd or tt tdtd, gray). The objective of this study was to investigate the nature of flavonoids in the pubescence of near-isogenic lines (NILs) for these loci. Flavonoids were extracted with methanol from pubescence of cultivar Clark with tawny pubescence (TT TdTd) and its NILs; from Clark-t with gray pubescence (tt TdTd) and Clark-td with near-gray pubescence (TT tdtd); and from a pair of NILs, To7B with tawny (TT TdTd) and To7G with gray pubescence (tt TdTd). Primary flavonoids were flavone aglycones. Luteolin and apigenin were predominant in NILs with tawny and gray pubescence, respectively. Small amount of 7-O-glucosides of the 2 flavones were also detected. Alleles at T locus were associated with 3'-hydroxylation in the B-ring of the flavones. The primary flavonoids in Clark-td were luteolin similar to Clark, but its amount was halved. High performance liquid chromatography peaks probably corresponding to isoflavonoids were found only in Clark-td in 2003. However, the peaks were not observed in 2005. The above results suggest that Td may encode a structural or a regulatory gene controlling flavone biosynthesis. Pigments remained visible in pubescence after methanol extraction, suggesting that a major part of the pigments was polymerized. Surface rinsing experiments revealed that flavone aglycones exist outside the surface of cells.     

On-line identification of further flavone C- and O-glycosides from sugarcane (Saccharum officinarum L., Gramineae) by HPLC-UV-MS

HPLC-UV-MS (APCI-MS/MS and CID/MS) was utilised for the identification of eight additional flavone glycosides from sugarcane (Saccharum officinarum L., Gramineae) extracts of bagasse, leaves and juice ('garapa'). Relevant information about substitution patterns was obtained through UV detection using post-column addition of shift reagents, while tandem MS provided structural information confirming the proposed structures of the C-glycosides vitexin, orientin, luteolin-8-C-(rhamnosylglucoside), 4',5'-dimethyl-luteolin-8-C-glycoside and the isomeric pair schaftoside-isoschaftoside, besides the O-glycosides tricin-7-O-neohesperidoside and tricin-7-O-glycoside.     

Anticlastogenic effect of flavonoids against mutagen-induced micronuclei in mice.

14 flavonoids, including flavone and flavonol derivatives, were tested for their anticlastogenic effect against induction of micronuclei by benzo[a]pyrene in polychromatic erythrocytes of mice. When each flavonoid was administered orally, together with intraperitoneally administered benzo[a]pyrene, most flavonol derivatives showed an anticlastogenic effect. The data suggest that the 2,3-double bond and 3,5,7-hydroxyl groups in the flavonoid molecules may be essential to produce anticlastogenic effects against benzo[a]pyrene. Galangin, one of the active compounds, and (-)-epicatechin, a weak one, were administered to mice in order to compare their anticlastogenic effect against 3 different kinds of carcinogens: ethyl methanesulfonate, 7,12-dimethylbenz[a]anthracene, and adriamycin. Galangin showed a stronger anticlastogenic effect than (-)-epicatechin against ethyl methanesulfonate and 7,12-dimethylbenz[a]anthracene. However, there was no significant effect against adriamycin-induced micronuclei by both compounds. Our study indicates that most flavonoids are anticlastogenic agents. Their anticlastogenic effects are apparently independent of their own clastogenic activities. Furthermore, their anticlastogenic activities do not apply universally to all types of genotoxic chemicals.     

植物(口山)酮甙类化合物

本文对目前已发现的天然产(口山)酮甙类化合物及其主要植物来源进行了综述,重点介绍了(口山)酮碳甙和氧甙的结构特点,同时介绍了它们在植物界的分布情况以及提取分离方法和药理学研究进展。     

Effects of flavonoids on nonenzymatic lipid peroxidation: structure-activity relationship

The in vitro effects of several flavonoids on nonenzymatic lipid peroxidation in the rat brain mitochondria was studied. The lipid peroxidation was indexed by measuring the MDA production using the 2-thiobarbituric acid TBA test. The flavonoids, apigenin, flavone, flavanone, hesperidin, naringin, and tangeretin promoted the ascorbic acid-induced lipid peroxidation, the extent of which depended upon the concentration of the flavonoid and ascorbic acid. The other flavonoids studied, viz., quercetin, quercetrin, rutin, taxifolin, myricetin, myricetrin, phloretin, phloridzin, diosmetin, diosmin, apiin, hesperetin, naringenin, (+)-catechin, morin, fisetin, chrysin, and 3-hydroxyflavone, all showed varying extents of inhibition of the nonenzymatic lipid peroxidation, induced by either ascorbic acid or ferrous sulfate. The flavonoid aglycones were more potent in their antiperoxidative action than their corresponding glycosides. Structure-activity analysis revealed that the flavonoid molecule with polyhydroxylated substitutions on rings A and B, a 2,3-double bond, a free 3-hydroxyl substitution and a 4-keto moiety, would confer upon the compound potent antiperoxidative properties.     

Chemosystematics in the Rutaceae: A chemotaxonomic survey of flavonoids and monoterpenes in leaves of Acmadenia species

By means of thin layer chromatography and gas liquid chromatography the distribution of monoterpenes and flavonoid aglycones in the leaves of 19 samples of 15 species of Acmadenia has been investigated. The distribution of flavonol and flavone aglycones shows a close agreement with recently proposed taxonomic divisions within the genus. It is proposed that these patterns may be of importance in interpreting the evolutionary development of the genus.     

Comprehensive profiling and natural variation of flavonoids in rice

Flavonoids constitute a major group of plant phenolic compounds. While extensively studied in Arabidopsis, profiling and natural y occurring variation of these compounds in rice (Oryza sativa), the mon...     

Mechanisms involved in the chemoprevention of flavonoids

Flavonoids, widespread in edible plants, have been studied extensively for their anticarcinogenic properties. However, only few studies have been done with these constituents being administered by the dietary route. In our research, the effects of feeding rats with flavone, flavanone, tangeretin, and quercetin were investigated on two steps of aflatoxin B1 (AFB1)-induced hepatocarcinogenesis (initiation and promotion). Nonpolar flavonoids such as flavone, flavanone and tangeretin administered through the initiation period, decreased the number of -gamma-glutamyl transpeptidase-preneoplastic foci. In the same conditions of administration, quercetin, a polyhydroxylated flavonoid, showed no protective effect. Moreover, feeding rats with flavanone during the phenobarbital-induced promotion step significantly reduced the areas of placental glutathione S-transferase preneoplastic foci. Quercetin, flavone, and tangeretin, administered in the same conditions, caused no significant effect. Therefore flavanone act as an anti-initiator as well as an anti-promotor. Several mechanisms were involved in the anti-initiating effects of flavone, flavanone, and tangeretin: enhancement of enzymes involved in the detoxication of AFB1 (glutathione S-transferase, UDP-glucuronyl transferase), increase of the formation of AFB1-glutathione conjugates and inhibition of the binding of AFB1 to DNA. Although the relevance of these data to the human situation remains to be demonstrated, they confirm that several flavonoids administered by the dietary route possess promising chemoprotective effects.     

Isolation and HPLC quantitative analysis of antioxidant flavonoids from Alternanthera tenella Colla

Phytochemical analysis of the antioxidant ethanolic extract of Alternanthera tenella Colla led to the isolation of six flavonoids, acacetin 8-C-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranoside] (1), 2"-O-alpha-L-rhamnopyranosyl-vitexin (2), 2"-O-beta-D-glucopyranosyl-vitexin (3), vitexin (4), quercetin (5) and kaempferol (6). All the structures were established by ESI-MS and NMR spectroscopic methods. Antioxidant capacity of extract, fractions and isolated compounds was determined using the oxygen radical absorbance capacity (ORAC) assay and extract, fractions and flavonoids isolated showed antioxidant activity in vitro. Moreover, the total soluble phenolic contents of the extract and fractions were measured using the Folin-Ciocalteau reagent and the quantitative analysis of flavone C-glycosides major constituents was performed by HPLC.     

Feeding deterrency of flavonoids and related phenolics towards Schizaphis graminum and Myzus persicae: Aphid feeding deterrents in wheat

A number of naturally occurring flavonoids have been tested for their feeding deterrent activity against two aphid species, Schizaphis graminum and Myzus persicae. Most flavonoids, including a number of dihydrochalcones related to phloretin, showed strong deterrency at concentrations well within the range often found in plants. Flavanone and flavone glycosides showed weak feeding deterrency relative to their corresponding aglycones. S. graminum and M. persicae responded similarly towards the compounds tested. The feeding deterrency of wheat extracts towards S. graminum was confined to the phenolic fraction, which included the flavone tricin. The more polar phenolic fraction showed the strongest feeding deterrency towards S. graminum.     

罗汉果叶中黄酮甙元的研究

以罗汉果叶为原料提取出黄酮,黄酮经水解、溶剂萃取、柱层析等手段,得到高纯度甙元;经TLC、液相色谱、红外光谱、核磁共振法等分析手段进行结构鉴定,证实甙元分别为山奈酚、槲皮素。     

Acetophenone C-glucosides and stilbene O-glucosides in Upuna borneensis

Three acetophenone C-glycosides; 2,4,6-trihydroxyacetophenone 3-C-beta-(2'-O-p-hydroxybenzoyl)-glucopyranoside, 2,4,6-trihydroxyacetophenone 3-C-beta-(2'-O-E-coumaroyl)-glucopyranoside, 2,4,6-trihydroxyacetophenone 3-C-beta-(2'-O-E-cinnamoyl)-glucopyranoside, and two resveratrol O-glycosides; piceid 2'-O-p-hydroxybenzoate and, piceid 2'-O-E-ferulate, together with three known compounds were isolated from the acetone soluble part of stem of Upuna borneensis (Dipterocarpaceae). The structures of isolates were determined by spectral analysis including extensive 2D-NMR spectral analyses.     

Isolation and identification of flavonoids, including flavone rotamers, from the herbal drug 'Crataegi folium cum flore' (hawthorn)

Twelve flavonoids, including seven flavones, four flavonols and one flavanone, were isolated from methanolic extract of the herbal drug 'Crataegi folium cum flore' (hawthorn leaves and flowers) by a combination of CC (over Amberlite XAD-7 and Sephadex LH-20) and preparative HPLC. Their structures, including that of the novel flavonol 8-methoxykaempferol 3-O-(6"-malonyl-beta-glucopyranoside), were elucidated by homo- and heteronuclear NMR and electrospray/MS. The 1H- and 13C-NMR of all compounds, including rotameric pairs of five flavone C-glycosides, were assigned. The presence and relative proportion of each rotamer was shown by various NMR experiments, including two-dimensional nuclear Overhauser and exchange spectroscopy, to depend on solvent, linkage position and structure of the C-glycosyl substituent.     

Flavones and flavone synthases

Within the secondary metabolite class of flavonoids which consist of more than 9000 known structures, flavones define one of the largest subgroups. Their natural distribution is demonstrated for almost all plant tissues. Various flavone aglyca and their O- or C-glycosides have been described in the literature. The diverse functions of flavones in plants as well as their various roles in the interaction with other organisms offer many potential applications, not only in plant breeding but also in ecology, agriculture and human nutrition and pharmacology. In this context, the antioxidative activity of flavones, their use in cancer prevention and treatment as well as the prevention of coronary heart disease should be emphasized. The therapeutic potential of flavones makes these compounds valuable targets for drug design, including recombinant DNA approaches. The biosynthesis of flavones in plants was found to be catalyzed by two completely different flavone synthase proteins (FNS), a unique feature within the flavonoids. The first, FNS I, a soluble dioxygenase, was only described for members of the Apiaceae family so far. The second, FNS II, a membrane bound cytochrome P450 enzyme, has been found in all other flavone accumulating tissues. This phenomenon is particularly of interest from the evolutionary point of view concerning the flavone biosynthesis and functions in plants. Recently, FNS I and FNS II genes have been cloned from a number of plant species. This now enables detailed biochemical and molecular characterizations and also the development of direct metabolic engineering strategies for modifications of flavone synthesis in plants to improve their nutritional and/or biopharmaceutical value.