Characterization of anthocyanins in grape juices by ion trap liquid chromatography-mass spectrometry

A reverse phase HPLC and electrospray interface with ion trap mass spectrometer method was developed for the characterization of anthocyanins in Concord, Rubired, and Salvador grape juices. Rubired and Salvador grapes are hybrids from Vitis vinifera and Vitis rupestris. Concord grape is a grape from the native American cultivar Vitis labrusca. Individual anthocyanins in these three varieties were identified on the basis of UV-vis and MS spectra and further elucidated by MS/MS spectra. Anthocyanins in Salvador and Concord grapes were 3-O-glucosides, 3-O-(6' '-O-p-coumaroyl)glucosides, 3-O-(6' '-O-p-acetyl)glucosides, 3,5-O-diglucosides, and 3-O-(6' '-O-p-coumaroyl)-5-O-diglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. Vitisin B was detected in Salvador grape juice. Anthocyanins in Rubired grape juice were primarily anthocyanin diglucosides: peonidin 3,5-O-diglucoside, malvidin 3,5-O-diglucoside, peonidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside, and malvidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside are the four major anthocyanins. The presence of pelargonidin 3-O-glucoside, not previously reported, has been established for the first time in all three juices.     

alpha-Glucosidase inhibitory action of natural acylated anthocyanins. 2. alpha-Glucosidase inhibition by isolated acylated anthocyanins

Four diacylated pelargonidin (Pg: SOA-4 and SOA-6), cyanidin (Cy: YGM-3), and peonidin (Pn: YGM-6) 3-sophoroside-5-glucosides isolated from the red flowers of the morning glory, Pharbitis nil cv. Scarlett O'Hara (SOA), and the storage roots of purple sweet potato, Ipomoea batatas cv. Ayamurasaki (YGM), were subjected to an alpha-glucosidase (AGH) inhibitory assay, in which the assay was performed with the immobilized AGH (iAGH) system to mimic the membrane-bound AGH at the small intestine. As a result, the acylated anthocyanins showed strong maltase inhibitory activities with IC(50) values of <200 microM, whereas no sucrase inhibition was observed. Of these, SOA-4 [Pg 3-O-(2-O-(6-O-(E-3-O-(beta-D-glucopyranosyl)caffeyl)-beta-D-glucopyranosyl)-6-O-E-caffeyl-beta-D-glucopyranoside)-5-O-beta-D-glucopyranoside] possessed the most potent maltase inhibitory activity (IC(50) = 60 microM). As a result of a marked reduction of iAGH inhibitory activity by deacylating the anthocyanins, that is, Pg (or Cy or Pn) sophoroside-5-glucoside, acylation of anthocyanin with caffeic (Caf) or ferulic (Fer) acid was found to be important in the expression of iAGH (maltase) inhibition. In addition, the result that Pg-based anthocyanins showed the most potent maltase inhibition, with an IC(50) value of 4.6 mM, and the effect being in the descending order of potency of Pg > Pn/Cy strongly suggested that no replacement at the 3'(5')-position of the aglycon B-ring may be essential for inhibiting iAGH action.     

Determination of anthocyanidins in berries and red wine by high-performance liquid chromatography

A high-performance liquid chromatographic (HPLC) method for the determination of anthocyanidins from berries and red wine is described. Delphinidin, cyanidin, petunidin, pelargonidin, peonidin, and malvidin contents of bilberry (Vaccinium myrtillus), black currant (Ribes nigrum), strawberry (Fragaria ananassa cv. Jonsok), and a Cabernet sauvignon (Vitis vinifera) red wine were determined. The aglycon forms of the anthocyanins present in the samples were revealed by acid hydrolysis. A reversed phase analytical column was employed to separate the anthocyanidins before identification by diode array detection. The suitability of the method was tested by determining the recovery (95-102% as aglycons and 69-104% from glycosides) for each anthocyanidin. Method repeatability was tested by charting the total aglycon content of two samples over a period of 14 analyses and determining the coefficients of variation (1.41% for bilberry and 2.56% for in-house reference material). The method developed proved thus to be effective for reliable determination of anthocyanidins from freeze-dried berry samples and red wine. The total anthocyanidin content of the tested samples was as follows: in-house reference material, 447 +/- 8 mg/100 g; strawberry, 23.8 +/- 0.4 mg/100 g; black currant, 135 +/- 3 mg/100 g; bilberry, 360 +/- 3 mg/100 g; and Cabernet sauvignon red wine, 26.1 +/- 0.1 mg/100 mL.     

Onions: a source of unique dietary flavonoids

Onion bulbs (Allium cepa L.) are among the richest sources of dietary flavonoids and contribute to a large extent to the overall intake of flavonoids. This review includes a compilation of the existing qualitative and quantitative information about flavonoids reported to occur in onion bulbs, including NMR spectroscopic evidence used for structural characterization. In addition, a summary is given to index onion cultivars according to their content of flavonoids measured as quercetin. Only compounds belonging to the flavonols, the anthocyanins, and the dihydroflavonols have been reported to occur in onion bulbs. Yellow onions contain 270-1187 mg of flavonols per kilogram of fresh weight (FW), whereas red onions contain 415-1917 mg of flavonols per kilogram of FW. Flavonols are the predominant pigments of onions. At least 25 different flavonols have been characterized, and quercetin derivatives are the most important ones in all onion cultivars. Their glycosyl moieties are almost exclusively glucose, which is mainly attached to the 4', 3, and/or 7-positions of the aglycones. Quercetin 4'-glucoside and quercetin 3,4'-diglucoside are in most cases reported as the main flavonols in recent literature. Analogous derivatives of kaempferol and isorhamnetin have been identified as minor pigments. Recent reports indicate that the outer dry layers of onion bulbs contain oligomeric structures of quercetin in addition to condensation products of quercetin and protocatechuic acid. The anthocyanins of red onions are mainly cyanidin glucosides acylated with malonic acid or nonacylated. Some of these pigments facilitate unique structural features like 4'-glycosylation and unusual substitution patterns of sugar moieties. Altogether at least 25 different anthocyanins have been reported from red onions, including two novel 5-carboxypyranocyanidin-derivatives. The quantitative content of anthocyanins in some red onion cultivars has been reported to be approximately 10% of the total flavonoid content or 39-240 mg kg (-1) FW. The dihydroflavonol taxifolin and its 3-, 7-, and 4'-glucosides have been identified in onions. Although the structural diversity of dihydroflavonols characterized from onions is restricted compared with the wide structural assortment of flavonols and anthocyanins identified, they may occur at high concentrations in some cultivars. From bulbs of the cultivar "Tropea", 5.9 mg of taxifolin 7-glucoside and 98.1 mg of taxifolin have been isolated per kilogram of FW.     

Insulin secretion by bioactive anthocyanins and anthocyanidins present in fruits

Anthocyanins are responsible for a variety of bright colors including red, blue, and purple in fruits, vegetables, and flowers and are consumed as dietary polyphenols. Anthocyanin-containing fruits are implicated in a decrease in coronary heart disease and are used in antidiabetic preparations. In the present study, we have determined the ability of anthocyanins, cyanidin-3-glucoside (1), delphinidin-3-glucoside (2), cyanidin-3-galactoside (3), and pelargonidin-3-galactoside (4), and anthocyanidins, cyanidin (5), delphinidin (6), pelargonidin (7), malvidin (8), and petunidin (9), to stimulate insulin secretion from rodent pancreatic beta-cells (INS-1 832/13) in vitro. The compounds were tested in the presence of 4 and 10 mM glucose concentrations. Our results indicated that 1 and 2 were the most effective insulin secretagogues among the anthocyanins and anthocyanidins tested at 4 and 10 mM glucose concentrations. Pelargonidin-3-galactoside is one of the major anthocyanins, and its aglycone, pelargonidin, caused a 1.4-fold increase in insulin secretion at 4 mM glucose concentration. The rest of the anthocyanins and anthocyanidins tested in our assay had only marginal effects on insulin at 4 and 10 mM glucose concentrations.     

Identification of antioxidative flavonols and anthocyanins in Sicana odorifera fruit peel

Ten flavonols and three anthocyanins were identified in the fruit peel of melo?n de olor (Sicana odorifera), and their structures were established by spectrometric and spectroscopic (ESI-MS and NMR) techniques. One of the identified flavonols, quercetin 3-O-(6'-O-malonyl)-β-D-glucopyranoside 4'-O-β-D-glucopyranoside, has not been reported before in the plant kingdom. Although quercetin-3-O-α-L-rhamnopyranosyl-(1→6)-β-d-glucopyranoside-4'-O-β-D-glucopyranoside had been reported before in literature and structure elucidation was done by comparison of NMR data with published data, to the best of our knowledge complete 1D and 2D NMR data have not been not delineated so far. Moreover, the antioxidant activity of pure compounds was measured by ABTS assay. It was established that quercetin 3-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside, quercetin-3-O-(6'-malonyl)-glucopyranoside, quercetin-3-O-β-D-glucopyranoside, and quercetin-3-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside-4'-O-β-D-glucopyranoside contribute significantly to the antioxidant activity exhibited by the fruit peel methanolic extract.     

Relationship between the biological activities of methylated derivatives of (-)-epigallocatechin-3-O-gallate (EGCG) and their cell surface binding activities

It was previously reported that (-)-epigallocatechin-3-O-gallate (EGCG) suppresses the expression of the high-affinity IgE receptor FcepsilonRI in human basophilic cells and that this suppressive effect is associated with EGCG binding to the cell surface. This study examined the effects of five methylated derivatives of EGCG, (-)-epigallocatechin-3-O-(3-O-methyl)gallate (EGCG 3' 'Me), (-)-epigallocatechin-3-O-(4-O-methyl)gallate (EGCG 4' 'Me), (-)-4'-O-methyl-epigallocatechin-3-O-gallate (EGCG 4'Me), (-)-epigallocatechin-3-O-(3,4-O-methyl)gallate (EGCG 3' '4' 'diMe), and (-)-4'-O-methyl-epigallocatechin-3-O-(4-O-methyl)gallate (EGCG 4'4' 'diMe) on FcepsilonRI expression and ERK1/2 phosphorylation, and each of their cell surface binding activities was measured. Of these five methylated derivatives, three that are methylated at the 3' '- and/or 4' '-position, EGCG 3' 'Me, EGCG 4' 'Me, and EGCG 3' '4' 'diMe, suppressed FcepsilonRI expression and ERK1/2 phosphorylation, although the suppressive effects were lower than that of EGCG. EGCG 4'Me and EGCG 4'4' 'diMe, both of which are methylated at the 4'-position, did not demonstrate a suppressive effect. Furthermore, it was found that EGCG 3' 'Me, EGCG 4' 'Me, EGCG 3' '4' 'diMe, and EGCG 4'Me, which are methylated at the 3' '- and/or 4' '-positions or the 4'-position, could bind to the cell surface even though their binding activities were lower than that of EGCG. Only EGCG 4'4' 'diMe, which is methylated at both the 4'- and 4' '-positions, could not bind. These results suggest that the trihydroxyl structure of the B ring is essential for EGCG to exert the suppressive effects and that the hydroxyl groups on both the 4'-position in the B ring and the 4' '-position in the gallate are crucial for the cell surface binding activity of EGCG.     

Identification of cyanidin 3-O-β-(6″-(3-hydroxy-3-methylglutaroyl)glucoside) and other anthocyanins from wild and cultivated blackberries

Anthocyanins from blackberries are natural dietary pigments. The aim of this study was to investigate the occurrence of anthocyanins in fruits of wild Norwegian blackberries and three blackberry ( Rubus fruticosus L.) cultivars and to report the complete identification of cyanidin 3-O-β-(6″-(3-hydroxy-3-methylglutaroyl)glucopyranoside), 5. This new pigment is most probably the same pigment that has previously been reported to occur in various blackberry samples as cyanidin 3-dioxalylglucoside. All of the examined blackberry samples contained in similar relative proportions the 3-glucoside (1), 3-rutinoside (2), 3-xyloside (3), and 3-O-β-(6″-malonylglucoside) (4) of cyanidin and 5. The absolute amounts of 1-5 in the wild Norwegian blackberries were 249, 18, 10, 24, and 22 mg of cyanidin 3-glucoside equivalents/100 g of fresh weight, respectively.     

Flavonoids in monospecific eucalyptus honeys from Australia

The HPLC analyses of Australian unifloral Eucalyptus honeys have shown that the flavonoids myricetin (3,5,7,3',4', 5'-hexahydroxyflavone), tricetin (5,7,3',4',5'-pentahydroxyflavone), quercetin (3,5,7,3',4'-pentahydroxyflavone), luteolin (5,7,3', 4'-tetrahydroxyflavone), and kaempferol (3,5,7, 4'-tetrahydroxyflavone) are present in all samples. These compounds were previously suggested as floral markers of European Eucalyptus honeys. The present results confirm the use of flavonoid analysis as an objective method for the botanical origin determination of eucalyptus honey. Honeys from E. camaldulensis (river red gum honey) contain tricetin as the main flavonoid marker, whereas in honeys from E. pilligaensis (mallee honey), luteolin is the main flavonoid marker, suggesting that species-specific differences can be detected with this analysis. The main difference between the flavonoid profiles of Australian and European Eucalyptus honeys is that in the Australian honeys, the propolis-derived flavonoids (pinobanksin (3,5, 7-trihydroxyflavanone), pinocembrin (5,7-dihydroxyflavanone), and chrysin (5,7-dihydroxyflavone)) are seldom found and in much smaller amounts.     

Identification of 5,7,3',4'-tetramethoxyflavone metabolites in rat urine by the isotope-labeling method and ultrahigh-performance liquid chromatography-electrospray ionization-mass spectrometry

5,7,3',4'-Tetramethoxyflavone (TMF), one of the major polymethoxyflavones (PMFs) isolated from Kaempferia parviflor , has been reported possessing various bioactivities, including antifungal, antimalarial, antimycobacterial, and anti-inflammatory activities. Although several studies on the TMF have been reported, the information about the metabolism of TMF and the structures of TMF metabolites is still not yet clear. In this study, an isotope-labeling method was developed for the identification of TMF metabolites. Three isotope-labeled TMFs (5,7,3',4'-tetramethoxy[3'-D(3)]flavone, 5,7,3',4'-tetramethoxy[4'-D(3)]flavone, and 5,7,3',4'-tetramethoxy[5,4'-D(6)]flavone) were synthesized and administered to rats. The urine samples were collected, and the main metabolites were monitored by ultrahigh-performance liquid chromatography-electrospray ionization-mass spectrometry. Five TMF metabolites were unambiguously identified as 3'-hydroxy-5,7,4'-trimethoxyflavone, 7-hydroxy-5,3',4'-trimethoxyflavone sulfate, 7-hydroxy-5,3',4'-trimethoxyflavone, 4'-hydroxy-5,7,3'-trimethoxyflavone, and 5-hydroxy-7,3',4'-trimethoxyflavone.     

Blackberry anthocyanins are mainly recovered from urine as methylated and glucuronidated conjugates in humans

The consumption of anthocyanins has been shown to prevent certain chronic diseases. However, anthocyanin metabolism has not yet been fully elucidated. The aim of this study was to evaluate anthocyanin urinary excretion in humans receiving a meal containing blackberries and to identify possible metabolites in urine. Five healthy volunteers were fed 200 g of blackberries (960 mumol of anthocyanins). Urine samples were collected and rapidly treated by solid-phase extraction. Anthocyanin metabolites were identified and quantified by HPLC-ESI-MS-MS and HPLC with UV-vis detection, respectively. In addition to native cyanidin 3-glucoside, several other anthocyanin metabolites were identified in the urine: methylated glycosides, glucuronides of anthocyanidins and anthocyanins, a sulfoconjugate of cyanidin, and anthocyanidins. Total urinary excretion of blackberry anthocyanin metabolites was 0.160 +/- 0.020% (n = 5) of the amount of anthocyanins ingested. Monoglucuronides of anthocyanidins represented >60% of this excretion. Urinary excretion of anthocyanins was maximal between 2 and 4 h after the meal, but continued during the 24 h of the experiment. This study highlighted the influence of aglycon structure on anthocyanin urinary excretion. It demonstrated that anthocyanins are not only methylated but also glucuroconjugated and sulfoconjugated in humans and that the main metabolites of blackberry anthocyanins in human urine were anthocyanidin monoglucuronides.     

Profiling glucosinolates and phenolics in vegetative and reproductive tissues of the multi-purpose trees Moringa oleifera L. (horseradish tree) and Moringa stenopetala L

Moringa species are important multi-purpose tropical crops, as human foods and for medicine and oil production. There has been no previous comprehensive analysis of the secondary metabolites in Moringa species. Tissues of M. oleifera from a wide variety of sources and M. stenopetala from a single source were analyzed for glucosinolates and phenolics (flavonoids, anthocyanins, proanthocyanidins, and cinnamates). M. oleifera and M. stenopetala seeds only contained 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate at high concentrations. Roots of M. oleifera and M. stenopetala had high concentrations of both 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate and benzyl glucosinolate. Leaves from both species contained 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate and three monoacetyl isomers of this glucosinolate. Only 4-(alpha-l-rhamnopyranosyloxy)-benzylglucosinolate was detected in M. oleifera bark tissue. M. oleifera leaves contained quercetin-3-O-glucoside and quercetin-3-O-(6' '-malonyl-glucoside), and lower amounts of kaempferol-3-O-glucoside and kaempferol-3-O-(6' '-malonyl-glucoside). M. oleifera leaves also contained 3-caffeoylquinic acid and 5-caffeoylquinic acid. Leaves of M. stenopetala contained quercetin 3-O-rhamnoglucoside (rutin) and 5-caffeoylquinic acid. Neither proanthocyanidins nor anthocyanins were detected in any of the tissues of either species.     

Properties of 3-deoxyanthocyanins from sorghum

There is increasing interest in natural food colorants with functional properties. Anthocyanins from black, brown (containing tannins), and red sorghums were characterized by spectrophotometric and HPLC techniques. The antioxidant activity and pH stability of the anthocyanins were also determined. Sorghum brans had 3-4 times higher anthocyanin contents than the whole grains. Black sorghum had the highest anthocyanin content (average = 10.1 mg/g in bran). The brown and red sorghum brans had anthocyanin contents of 2.8-4.3 mg/g. Only 3-deoxyanthocyanidins were detected in sorghum. These compounds are more stable to pH-induced color change than the common anthocyanidins and their glycosides. Additionally, crude sorghum anthocyanin extracts were more stable than the pure 3-deoxyanthocyanidins. The antioxidant properties of the 3-deoxyanthocyanidins were similar to those of the anthocyanins. Pigmented sorghum bran has high levels of unique 3-deoxyanthocyanidins, which are stable to change in pH and have a good potential as natural food pigments.     

Identification of flavonoids in Hakmeitau beans (Vigna sinensis) by high-performance liquid chromatography-electrospray mass spectrometry (LC-ESI/MS)

Liquid chromatography coupled with electrospray mass spectrometry (LC-ESI/MS) with positive and negative ion detection was used for the identification of flavonoids in Hakmeitau beans, a black seed cultivar of cowpea (Vigna sinensis). Gradient elution with water and acetonitrile, both containing 2% formic acid, was employed in chromatographic separation. The peaks were identified by comparison of the retention times and the UV-vis spectroscopic and mass spectrometric data with authentic standards and/or literature data. The identified flavonoids included six anthocyanins (cyanidin 3-O-galactoside, cyanidin 3-O-glucoside, delphinidin 3-O-glucoside, malvidin 3-O-glucoside, peonidin 3-O-glucoside, and petunidin 3-O-glucoside) and four flavonol/flavonol glycosides (kaempferol 3-O-glucoside, quercetin, quercetin 3-O-glucoside, and quercetin 3-O-6' '-acetylglucoside). The tentatively identified flavonoids included two anthocyanins (malvidin 3-O-acetylglucoside and peonidin 3-O-malonylglucoside) and three flavonol glycosides (myricetin-3-O-glucoside, quercetin 7-O-glucoside, and quercetin-3-O-diglucoside). These flavonoids are present in seed coats, and the contents of anthocyanins and flavonol glycosides were 20.7 and 2.0 mg/g, respectively.     

The flavonoids of tomatoes

Tomatoes ( Lycopersicon esculentum Mill.) have been recognized as an important source of dietary flavonoids because of a high consumption worldwide. The qualitative and quantitative flavonoid compositions of assorted tomato cultivars including individual quantitative contributions of the five most significant flavonoids have been determined in this work. The dihydrochalcone phloretin 3',5'-di-C-beta-glucopyranoside and the flavonol quercetin 3-O-(2'-O-beta-apiofuranosyl-6'-O-alpha-rhamnopyranosyl-beta-glucopyranoside) were identified for the first time in Solanaceae spp. and found to be among the main flavonoids in all cultivars. Phloretin 3',5'-di-C-glc is the first C-glycoside identified in tomatoes and also the first dihydrochalcone from this species. In addition, chalconaringenin, kaempferol 3-rutinoside, and quercetin 3-rutinoside (rutin), though previously reported to occur in tomato, were fully characterized by extensive use of 2D NMR techniques and high-resolution LCMS. The total flavonoid content of different tomato types varied from 4 to 26 mg 100 (-1) g FW with chalconaringenin as the predominant compound comprising 35 to 71% of the total flavonoid content. The individual quantities of quercetin 3-O-(2'- O-beta-apiofuranosyl-6'- O-alpha-rhamnopyranosyl-beta-glucopyranoside) and phloretin 3',5'-di-C-beta-glucopyranoside was similar to that of rutin in several cultivars.     

LC-ESI-MS study of the flavonoid glycoside malonates of red clover (Trifolium pratense)

High-performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was applied to the analysis of the flavonoids and their glycoside malonates of the flowers and leaves of red clover (Trifolium pratense). Through LC-MS comparative studies on the plant extracts and their malonate-free extracts, approximately 20 flavonoid glycoside malonates were detected in the flower extract. Eight were identified as genistin 6' '-O-malonate (39), formononetin 7-O-beta-D-glucoside 6' '-O-malonate (40), biochanin A 7-O-beta-D-glucoside 6' '-O-malonate (41), trifoside 6' '-O-malonate (42), irilone 4'-O-beta-D-glucoside 6' '-O-malonate (43), pratensein 7-O-beta-D-glucoside 6' '-O-malonate (44), isoquercitrin 6' '-O-malonate (45), and 3-methylquercetin 7-O-beta-D-glucoside 6' '-O-malonate (46). About 15 other flavonoids and clovamides were proved to be present in this extract. The study also found that the flowers contained flavones as the major flavonoids, whereas the leaves had isoflavones as the major flavonoids. This is the first detection of the six malonates (39 and 42-46) in the extracts of red clover, and among them, 42, 43, and 46 are new compounds.     

Antimutagens in gaiyou (Artemisia argyi levl. et vant.)

Antimutagens from gaiyou (Artemisia argyi Levl. et Vant., Compositae) were examined. The methanol extract prepared from aerial parts of this plant strongly reduced the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), when Salmonella typhimurium TA98 was used in the presence of the rat liver microsomal fraction. The antimutagens were purified chromatographically while monitoring the antimutagenic activity against Trp-P-2 with a modified Ames test employing a plate method. This purification resulted in the isolation of four strong antimutagens, 5,7-dihydroxy-6,3',4'-trimethoxyflavone (eupatilin), 5, 7,4'-trihydroxy-6,3'-dimethoxyflavone (jaceosidin), 5,7, 4'-trihydroxyflavone (apigenin) and 5,7, 4'-trihydroxy-3'-methoxyflavone (chrysoeriol) from the methanol extract. These antimutagenic flavones exhibited strong antimutagenic activity against not only Trp-P-2 but also against other heterocyclic amines, such as 3-amino-1,4-dimethyl-5H-pyrido[4, 3-b]indole (Trp-P-1), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3, 8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA(alpha)C) in S. typhimurium TA98. In contrast, they did not exhibit antimutagenic activity against benzo[a]pyrene (B[a]P), 4-nitroquinoline-1-oxide (4-NQO), 2-aminofluorene (2-AF), 2-nitrofluorene (2-NF) or furylfuramide (AF-2) in S. typhimurium TA98, or B[a]P, 4-NQO, 2-NF, AF-2, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or sodium azide (SA) in Salmonella typhimurium TA100, whereas they decreased the mutagenicity caused by aflatoxin B(1) (AFB(1)) and 2-aminoanthracene (2-AA) in both of these tester strains. Regarding the structure-activity relationship, the tested flavones had distinct differences in the intensities of their antimutagenic activities according to the differences of their substitution patterns. Namely, the intensity of antimutagenic activities against Trp-P-2 decreased in the order of: 5,7,3',4'-tetrasubstituted flavones (IC(50): <0.1 mmol/plate), 5,7,4'-trisubstituted flavones (IC(50): 0.120-0.260 mmol/plate), 5,6,7,3',4'-pentasubstituted flavones (IC(50): 0.440-0. 772 mmol/plate). The four isolated flavones were also studied regarding their antimutagenic mechanisms with preincubation methods of the modified Ames test and emission spectroscopic analysis. The results suggested that all isolated flavones were desmutagens which directly inactivated Trp-P-2 or inhibited its metabolic activation.     

Inhibition of lipid peroxidation and structure-activity-related studies of the dietary constituents anthocyanins,anthocyanidins, and catechins

The antioxidant activities of a series of commonly consumed and biogenetically related plant phenolics, namely, anthocyanidins, anthocyanins, and catechins, in a liposomal model system have been investigated. The antioxidant efficacies of the compounds were evaluated on their abilities to inhibit the fluorescence intensity decay of an extrinsic probe, 3-[p-(6-phenyl)-1,3,5-hexatrienyl]phenylpropionic acid, caused by free radicals generated during metal ion-induced peroxidation. Distinct structure-activity relationships were revealed for the antioxidant abilities of these structurally related compounds. Whereas antioxidant activity increased with an increasing number of hydroxyl substituents present on the B-ring for anthocyanidins, the converse was observed for catechins. However, substitution by methoxyl groups diminished the antioxidant activity of the anthocyanidins. Substitution at position 3 of ring C played a major role in determining the antioxidant activity of these classes of compounds. The anthocyanidins, which possess a hydroxyl group at position 3, demonstrated potent antioxidant activities. For the cyanidins, an increasing number of glycosyl units at position 3 resulted in decreased antioxidant activity. Similarly, the substitution of a galloyl group at position 3 of the flavonoid moiety resulted in significantly decreased antioxidant activity for the catechins. Among catechins, cis-trans isomerism, epimerization, and racemization did not play a role in overall antioxidant activity. The antioxidant activities of test compounds (at 40 microM concentrations) were compared to the commercial antioxidants tert-butylhydroquinone, butylated hydroxytoluene, butylated hydroxyanisole, and vitamin E (all at 10 microM concentrations).     

Studies on the constituents of Chenopodium quinoa seeds: isolation and characterization of new triterpene saponins

Six triterpenoid saponins were isolated from the edible grain quinoa, which is seeds of Chenopodium quinoa (Chenopodiaceae). Following are their structures: phytolaccagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (1); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (2); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' "-->3' ')-beta-D-xylopyranosyl-(1' '-->2')-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (3); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' "-->2' ')-beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (4); oleanolic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (5); and oleanolic acid 3-O-[beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (6). The oleanane-type saponins (5, 6) were isolated for the first time in this plant, two of the phytolaccagenane (1, 3) were new compounds and two (2, 4) were previously found in quinoa. The structures were characterized on the basis of hydrolysis and spectral evidence, including 1D- and 2-D NMR (HMQC and HMBC) and ESI-MS analyses.     

Structural analysis of a novel antimutagenic compound, 4-Hydroxypanduratin A, and the antimutagenic activity of flavonoids in a Thai spice, fingerroot (Boesenbergia pandurata Schult.) against mutagenic heterocyclic amines.

Six compounds were isolated from fresh rhizomes of fingerroot (Boesenbergia pandurata Schult.) as strong antimutagens toward 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) in Salmonella typhimurium TA98. These compounds were 2',4',6'-trihydroxychalcone (pinocembrin chalcone; 1), 2',4'-dihydroxy-6'-methoxychalcone (cardamonin; 2), 5,7-dihydroxyflavanone (pinocembrin; 3), 5-hydroxy-7-methoxyflavanone (pinostrobin; 4), (2,4,6-trihydroxyphenyl)-[3'-methyl-2'-(3' '-methylbut-2' '-enyl)-6'-phenylcyclohex-3'-enyl]methanone (5), and (2,6-dihydroxy-4-methoxyphenyl)-[3'-methyl-2'-(3' '-methylbut-2' '-enyl)-6'-phenylcyclohex-3'-enyl]methanone (panduratin A; 6). Compound 5 was a novel compound (tentatively termed 4-hydroxypanduratin A), and 1 was not previously reported in this plant, whereas 2-4 and 6 were known compounds. The antimutagenic IC(50) values of compounds 1-6 were 5.2 +/- 0.4, 5.9 +/- 0.7, 6.9 +/- 0.8, 5.3 +/- 1.0, 12.7 +/- 0.7, and 12.1 +/- 0.8 microM in the preincubation mixture, respectively. They also similarly inhibited the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). All of them strongly inhibited the N-hydroxylation of Trp-P-2. Thus, the antimutagenic effect of compounds 1-6 was mainly due to the inhibition of the first step of enzymatic activation of heterocyclic amines.