Chemotaxonomic consideration of flavonoids from the leaves of Chrysanthemum arcticum subsp. arcticum and yezoense,and related species

We examined the foliar flavonoids of Chrysanthemum arcticum subsp. arcticum and yezoense, and related Chrysanthemum species. Five flavonoid glycosides (luteolin 7-O-glucoside and 7-O-glucuronides of luteolin, apigenin, eriodictyol and naringenin) were isolated from these taxa. Luteolin 7-O-xylosylglucoside, luteolin, apigenin and quercetin 3-methyl ether were found in subsp. yezoense as very minor compounds that were not recognised by high-performance liquid chromatography/photodiode array (HPLC/PDA). The related species C. yezoense contained acacetin 7-O-rutinoside and some methoxylated flavone aglycones as major compounds. Thus, C. arcticum was distinguished from C. yezoense according to their flavonoid profiles.     

The flavonoids of local members of astereae (compositae)

Six species belonging to the genera Ceruana, Conyza, Conyzanthus and Grangea, tribe Astereae (Compositae), were investigated for their flavonoids. Glycosides identified were found to belong to the aglycones kaempferol, quercetin, luteolin, scutellarein and hispidulin. The chemosystematic relationships are discussed.     

Bioconversion of isoflavones during the fermentation of Samso-Eum with Lactobacillus strains

In this study, the possible application of Lactobacillus strains as a functional starter culture to ferment Samso-Eum (SE), an oriental herbal medicine formula, and the production of bioactive isoflavones (daidzein, genistein) were investigated. Four strains of Lactobacillus (Lactobacillus plantarum KFRI 144, L. amylophilus KFRI 161, L. curvatus KFRI 166, and L. bulgaricus KFRI 344) were used for SE fermentation. Declines in pH and in viable cell counts during fermentation were investigated and the quantification of isoflavones using HPLC were performed after fermentation at 37°C for 48 h. All the tested Lactobacillus strains lowered the pH level to approximately 3.6 after 48 h and showed the highest level of growth at 24 h during SE fermentation. During the SE fermentation of the four Lactobacillus strains, the conversion of isoflavone glycosides (daidzin, genistin) into bioactive aglycones (daidzein, genistein) was observed in all of the fermentations, but with different rates depending on the strains. L. plantarum KFRI 144 and L. amylophilus KFRI 161 exhibited the highest bioconversion rate of isoflavone glycosides into their bioactive aglycones. These results demonstrate that L. plantarum KFRI 144 and L. amylophilus KFRI 161 have potentials as functional starter cultures for manufacturing fermented SE with higher isoflavone bioavailability.     

External and internal flavonoids from Madagascarian Uncarina species (Pedaliaceae)

External and internal flavonoids were isolated from 12 Uncarina taxa (Pedaliaceae), endemic to Madagascar. Four flavone aglycones, tricetin 7,3′,5′-trimethyl ether, tricetin 7,4′,5′-trimethyl ether, 5,3′-dihydroxy-6,7,4′,5′-tetramethoxyflavone and eupatorin were isolated from leaf wax of seven Uncarina taxa, Uncarina grandidieri, Uncarina decaryi, Uncarina abbreviata, Uncarina turicana, Uncarina platycarpa, Uncarina leandrii var. leandrii and Uncarina peltata, but not Uncarina stellulifera, Uncarina perrieri, Uncarina sakalava, Uncarina leptocarpa and U. leandrii var. rechbergeri. Furthermore, eight flavonoid glycosides were isolated from the leaves. Major glycosides were apigenin and luteolin 7-O-glucuronides and occurred in all the Uncarina taxa examined, except the absence of the former compound in U. peltata. Other glycosides were identified as hispidulin, jaceosidin, chrysoeriol and tricin 7-O-glucuronides, and luteolin 7,4′-di-O-glucuronide and a flavonol, isorhamnetin 3-O-diglucoside. From the results described above, methylated flavone aglycones and glucuronides were chemical characters of the leaves of Uncarina species, and also may be those of the family Pedaliaceae. Besides, an anthocyanin, two flavonols and three flavones were isolated from the flowers of U. grandidieri, and identified as cyanidin 3-O-rutinoside (anthocyanin), quercetin and isorhamnetin 7-O-glucuronides (flavonols) and apigenin, luteolin and jaceosidin 7-O-glucuronides (flavones).     

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.     

A systematic study of the genus hulsea (Asteraceae)

A systematic treatment based on genetic, biochemical, and morphological studies is presented for the 11 recognized taxa ofHulsea. A uniform diploid complement of 38 chromosomes was found in the 37 populations examined, including all species and representing first reports forH. brevifolia, H. californica, andH. mexicana. Thirteen flavonoid compounds, based on the aglycones quercetin, apigenin, cyanidin, and the 4′-O-methyl ether of luteolin, were detected inHulsea. Patterns of the flavonoid compounds are discussed with respect to the systematic treatment. The results of 595 synthetic crosses indicate thatHulsea species are self-incompatible, relatively cross-compatible, and that their F₁ hybrids are relatively fertile. Heterozygosity for one or two translocations is reported for 11 interspecific and intersubspecific hybrid combinations.     

UDP-Glucose: Cyanidin 3-O-glucosyltransferase from red cabbage seedlings

An enzyme, catalysing the glucosylation of cyanidin at the 3-position using uridine diphosphate-D-glucose (UDPG) as glucosyl-donor, has been isolated and purified about 50-fold from young red cabbage (Brassica oleracea) seedlings. The pH optimum for this reaction was ca 8 and no additional cofactors were required. The reaction was inhibited by cyanidin (above 0.25 mM) and by very low concentrations of the reaction product cyanidin-3-glucoside (5 μM). The K_m values for UDPG and cyanidin were 0.51 and 0.4 mM respectively. In addition to cyanidin the enzyme could also glucosylate the following compounds at the 3-position: pelargonidin, peonidin, malvidin, kaempferol, quercetin, isorhamnetin, myricetin and fisetin. In contrast, cyanidin-3-glucoside, cyanidin-3-sophoroside, cyanidin-3,5-diglucoside, apigenin, luteolin, naringenin and dihydroquercetin were not glucosylated.     

Expression of flavonoid 6-hydroxylase candidate genes in normal and mutant soybean genotypes for glycitein content

Soybean seeds accumulate large amounts of isoflavones (genistein, daidzein and glycitein), secondary metabolites known for their phytoestrogenic activities. Isoflavone composition depends on the seed part and glycitein is almost found exclusively in hypocotyls. Moreover, two major phenotypes are encountered in soybean cultivars, with either low (35 %) or high (55 %) levels of glycitein in their hypocotyls. This trait was under a quasi-mendelian heredity, implicating at most one or two genes. A CYP71D9 cDNA displaying a flavonoid 6-hydroxylase (F6H) activity had previously been isolated from elicitor-induced soybean (Glycine max L.) cells. This enzyme allows the synthesis of the glycitein flavanone intermediate (6,7,4′- trihydroxyflavanone) by catalyzing the A-ring hydroxylation of liquiritigenin. In this study, the CYP71D9 gene (F6H1) and two other candidates (F6H2 and F6H3) were studied using contrasted soybean cultivars for glycitein content (0, 35, 55 and 80 %). Their expression was observed in chitosan elicited leaves. They encode P450 proteins of 496, 469 and 481 amino acids respectively and were expressed in leaves with or without elicitation. The expression patterns of these three genes were performed in cotyledons and hypocotyls at different developmental stages. F6H1 and F6H2 were not expressed in the developing seed. F6H3 was only expressed in hypocotyls. Its expression levels did not correlate with hypocotyls glycitein content, but it was not expressed in the null mutant for glycitein. Thus, this F6H3 gene is a good potential candidate for glycitein biosynthesis in soybean seed.     

Purification,structural elucidation,antioxidant capacity and neuroprotective potential of the main polyphenolic compounds contained in Achyrocline satureioides (Lam) D.C. (Compositae)

Achyrocline satureioides (Lam) D.C (Compositae) is a native medicinal plant of South America traditionally utilized for its anti-inflammatory, sedative and anti-atherosclerotic properties among others. Neuroprotective effects have been reported in vivo and could be associated to its elevated content of flavonoid aglycones. In the present study we performed the isolation and structure elucidation of the major individual flavonoids of A. satureioides along with the in vitro characterization of their individual antioxidant and neuroprotective properties in order to see their putative relevance for treating neurodegeneration.Exact mass, HPLC-MS/MS and ¹H NMR identified dicaffeoyl quinic acid isomers, quercetin, luteolin, isoquercitrin, and 3-O-methylquercetin as the mayor polyphenols. Flavonoids intrinsic redox properties were evaluated in the presence of the endogenous antioxidants GSH and Ascorbate. Density Functional Theory (DFT) molecular modeling and electron density studies showed a theoretical basis for their different redox properties. Finally, in vitro neuroprotective effect of each isolated flavonoid was evaluated against hydrogen peroxide-induced toxicity in a primary neuronal culture paradigm. Our results showed that quercetin was more efficacious than luteolin and isoquercitrin, while 3-O-methylquercetin was unable to afford neuroprotection significantly. This was in accordance with the susceptibility of each flavonoid to be oxidized and to react with GSH. Overall our results shed light on chemical and molecular mechanisms underlying bioactive actions of A. satureioides main flavonoids that could contribute to its neuroprotective effects and support the positive association between the consumption of A. satureioides as a natural dietary source of polyphenols, and beneficial health effect.     

High pressure liquid chromatographic analysis of flavonoid chemical markers in petals from Gerbera flowers as an adjunct for cultivar and germplasm identification

Flavonoids present in petals from Gerbera flowers were resolved and quantitated by high pressure liquid chromatography (HPLC). The anthocyanins isolated from 18 cultivars, ranging in color from orange through lavender, were pelargonidin and cyanidin 3-malonylglucosides accompanied by smaller amounts of pelargonidin and cyanidin 3-glucosides. Related flavonoid copigments were apigenin and luteolin 4′-glucosides and 7-glucosides, apigenin 7-malonylglucoside, kaempferol and quercetin 3-glucosides, 4′-glucosides and 3-malonylglucosides. Both qualitative and quantitative differences in these flavonoid chemical markers distinguished cultivars with very similar colors. Malonyl esters of anthocyanins are easily degraded by HCl and conventional extraction and purification procedures were adjusted to preserve their natural state.     

Effect of Circulating Forms of Soy Isoflavones on the Oxidation of Low Density Lipoprotein

Soy isoflavones are thought to have a cardioprotective effect that is partly mediated by an inhibitory influence on the oxidation of low density lipoprotein (LDL). However, the aglycone forms investigated in many previous studies do not circulate in appreciable quantities because they are metabolised in the gut and liver. We investigated effects of various isoflavone metabolites, including for the first time the sulphated conjugates formed in the liver and the mucosa of the small intestine, on copper-induced LDL oxidation. The parent aglycones inhibited oxidation, although only 5% as well as quercetin. Metabolism increased or decreased their effectiveness. Equol inhibited 2.65-fold better than its parent compound daidzein and 8-hydroxydaidzein, not previously assessed, was 12.5-fold better than daidzein. However, monosulphated conjugates of genistein, daidzein and equol were much less effective and disulphates completely ineffective. Since almost all isoflavones circulate as conjugates, these data suggest that despite the increased potency produced by some metabolic changes, isoflavones may not be effective antioxidants in vivo unless they are deconjugated again.     

Biotransformation of soybean isoflavones by a marine <Emphasis Type="Italic">Streptomyces</Emphasis> sp. 060524 and cytotoxicity of the products

A marine Streptomyces sp. 060524 capable of hydrolyzing the glycosidic bond of isoflavone glycosides, was isolated by detecting its β-glucosidase activity. 5 isoflavone aglycones were isolated from culture filtrates in soybean meal glucose medium. They were identified as genistein (1), glycitein (2), daidzein (3), 3′,4′,5,7-tetrahydroxyisoflavone (4), and 3′,4′,7-trihydroxyisoflavone (5), based on UV, NMR and mass spectral analysis. The Streptomyces can selectively hydroxylate at the 3′-position in the daidzein and genistein to generate 3′-hydroxydaidzein and 3′-hydroxygenistein, respectively. The Strain biotransformed more than 90% of soybean isoflavone glycosides into their aglycones within 108 h. 3′-hydroxydaidzein and 3′-hydroxygenistein exhibited stronger cytotoxicity against K562 human chronic leukemia than daidzein and genistein.     

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.     

Anti-obesity activities of fermented soygerm isoflavones by Bifidobacterium breve

Soygerm isoflavones were subjected to fermentation by Bifidobacterium breve. Most of isoflavone glycosides (daidzin, glycitin and genistin) in soygerms were deglycosylated to their corresponding isoflavone aglycones (daidzein, glycitein and genistein) within 24 h fermentation. Fermented isoflavones significantly inhibited pancreatic lipase activity in fermentation-time and dosage dependant manner. When fermented isoflavones were orally administered with olive oil to SD rats, the triglyceride (TG) level in plasma after 2 h of ingestion was significantly lower than the control of only olive oil administered group whereas no such significant decrease in plasma TG was observed in unfermented isoflavone administered group. This result indicates that oral administration of fermented isoflavones effectively suppressed absorption of excessive lipid into a body. Addition of either unfermented or fermented soygerm isoflavones effectively inhibited adipocyte differentiation from 3T3-L1 in a dose dependent manner. In conclusion, B. breve successfully converted soygerm isoflavones into their aglycones, and these aglycones were more effective in suppressing lipid absorption as well as adipocytes differentiation than their glycosides.     

A comparative study of the flavonoids of Medicago radiata with other Medicago and related Trigonella species

The flavonoid glycosides of Medicago radiata as well as M. arabica, M. polymorpha, M. sativa, Trigonella coerulescens, T. foenum-graecum and T. spicata were studied in detail. Major glycosides identified were the 7-glucuronides of apigenin, luteolin, chrysoeriol and tricin, as well as lesser amounts of di- and triglucuronides of chrysoeriol and tricin. Also identified were the 3-robinobioside and 3,7-diglucoside of kaempferol, along with lesser amounts of quercetin-3,7-diglucoside, 4′,7-dihydroxyflavone, 3′,4′,7-trihydroxyflavone, formononetin and daidzein. Twelve other Medicago and Trigonella species were also studied for their flavonoid aglycones. The systematic position of M. radiata is discussed.     

Development and validation of a RP-HPLC method for quantification of isoflavone aglycones in hydrolyzed soy dry extracts

Isoflavones are widely used as an alternative treatment to hormone replacement therapy and also for prevention of several chronic diseases, including cancers. Genistein, daidzein and glycitein are the most abundant isoflavone aglycones found in soy extracts, where they also occur as glycosides. This paper describes the development and validation of an isocratic reversed-phase HPLC (RP-HPLC) method for the analysis of isoflavone aglycones, released after acid hydrolysis of soy dry extracts, used as pharmaceutical raw material. The quantification was carried out in a C(18) endcapped column, using a mobile phase composed of 0.1% acetic acid and methanol (52:48), at a flow rate of 1.0 ml/min and diode array detection (DAD) at 254 nm. The method showed to be linear (r(2)>0.99), precise (R.S.D.<2%), accurate (recovery of 98.88% for daidzein and 98.12% for genistein), robust and specific.     

Flavonoid glycosides from illuminated cell suspension cultures of Petroselinum hortense

Twenty-four different flavonoid glycosides were isolated from illuminated cell suspension cultures of parsley (Petroselinum hortense). The chemical structures of fourteen of these compounds were further characterized. The aglycones identified were the flavones apigenin, luteolin and chrysoeriol, and the flavonols quercetin and isorhamnetin. The flavones occurred either as 7-O-glucosides or as 7-O-apioglucosides, while the flavonols were 3-O-monoglucosides or 3,7-O-diglucosides. One-half of these glycosides were electrophoretically mobile and substituted with malonate residues.     

Anaerobic Degradation of Flavonoids by Clostridium orbiscindens

An anaerobic, quercetin-degrading bacterium was isolated from human feces and identified as Clostridium orbiscindens by comparative 16S rRNA gene sequence analysis. The organism was tested for its ability to transform several flavonoids. The isolated C. orbiscindens strain converted quercetin and taxifolin to 3,4-dihydroxyphenylacetic acid; luteolin and eriodictyol to 3-(3,4-dihydroxyphenyl)propionic acid; and apigenin, naringenin, and phloretin to 3-(4-hydroxyphenyl)propionic acid, respectively. Genistein and daidzein were not utilized. The glycosidic bonds of luteolin-3-glucoside, luteolin-5-glucoside, naringenin-7-neohesperidoside (naringin), quercetin-3-glucoside, quercetin-3-rutinoside (rutin), and phloretin-2′-glucoside were not cleaved. Based on the intermediates and products detected, pathways for the degradation of the flavonol quercetin and the flavones apigenin and luteolin are proposed. To investigate the numerical importance of C. orbiscindens in the human intestinal tract, a species-specific oligonucleotide probe was designed and tested for its specificity. Application of the probe to fecal samples from 10 human subjects proved the presence of C. orbiscindens in 8 out of the 10 samples tested. The numbers ranged from 1.87 × 10⁸ to 2.50 × 10⁹ cells g of fecal dry mass⁻¹, corresponding to a mean count of 4.40 × 10⁸ cells g of dry feces⁻¹.     

Identification of isoflavones in Acca sellowiana and two Psidium species (Myrtaceae)

Several types of isoflavonoid-like immunoreactivity were detected in water-ethanolic extracts from Acca sellowiana (Berg) Burret, Psidium guajava L. and Psidium littorale Raddi (Myrtaceae) leaves. Chromatographic mobility of the immunoreactive substances was compared to that of authentic standards, revealing a spectrum of isoflavonoid metabolites in both genera. Aglycones as well as glycosides were detected, namely daidzin, genistin, daidzein, genistein, formononetin, biochanin A, prunetin, and several incompletely characterized isoflavones. Subsequent HPLC–MS study verified the identities of the main immunoreactive isoflavones and found several others, namely glycitein, glycitin, ononin, sissotrin, including the malonylated and acetylated glucosides. It is concluded that the isoflavonoid metabolic pathway is present in the Myrtaceae family.     

Structure-activity relationships of flavonoids for vascular relaxation in porcine coronary artery

Flavonoids are polyphenolic compounds that are widespread in the plant kingdom, and structure-activity relationships (SAR) for vascular relaxation effects were examined for 17 of them using porcine coronary arteries. Density functional theory was employed to calculate the chemical parameters of these compounds. The order of potency for vascular relaxation was as follows: flavones (apigenin and luteolin) >or= flavonols (kaempferol and quercetin)>isoflavones (genistein and daidzein)>flavanon(ol)es (naringenin)>chalcones (phloretin)>anthocyanidins (pelargonidin)>flavan(ol)es ((+)-catechin and (-)-epicatechin). SAR analysis revealed that for good relaxation activity, the 5-OH, 7-OH, 4'-OH, C2=C3 and C4=O functionalities were essential. Comparison of rutin with quercetin, genistin with genistein, and puerarin with daidzein demonstrated that the presence of a glycosylation group greatly reduced relaxation effect. Total energy and molecular volume were also predictive of their relaxation activities. Our findings indicated that the most effective relaxing agents are apigenin, luteolin, kaempferol and genistein. These flavonoids possess the key chemical structures demonstrated in our SAR analysis.