Characterization of anthocyanins and pyranoanthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juice

Anthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juices were isolated and purified by means of high-speed countercurrent chromatography and preparative high-performance liquid chromatography. Structures of the pigments were then elucidated by electrospray ionization multiple mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. The major anthocyanins of the juice were characterized as cyanidin 3-glucoside and cyanidin 3-(6"-malonylglucoside). Furthermore, six minor anthocyanins were detected and identified as cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, delphinidin 3-(6"-malonylglucoside), peonidin 3-(6"-malonylglucoside), and cyanidin 3-(6"-dioxalylglucoside). The occurrence of the latter compound in blood oranges is reported here for the first time, together with full NMR spectroscopic data. Further investigations revealed the presence of four anthocyanin-derived pigments, which are formed through a direct reaction between anthocyanins and hydroxycinnamic acids during prolonged storage of the juice. These novel pyranoanthocyanins were identified as the 4-vinylphenol, 4-vinylcatechol, 4-vinylguaiacol, and 4-vinylsyringol adducts of cyanidin 3-glucoside through comparison of their mass spectrometric and chromatographic properties with those of synthesized reference compounds.     

Seven new aminoacyl sugars in Ipomoea batatas

An analysis of the polar extracts from sweet potato Ipomoea batatas (Convolvulaceae) led to the isolation of seven unknown aminoacyl sugars. On the basis of 1D, 2D NMR, and mass spectrometry data, the structures of the compounds were elucidated as: beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-valyl]-glucopyranoside (1), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-tyrosyl]-glucopyranoside (2), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-threonyl]-glucopyranoside (3), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-hystidyl]-glucopyranoside (4), 2-beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-alanyl]-glucopyranoside (5), beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-tryptophanyl]-glucopyranoside (6), and beta-D-fructofuranosyl-(2 --> 1)-alpha-D-[2-O-glycyl]-glucopyranoside (7).     

Saponins in alfalfa (Medicago sativa L.) root and their structural elucidation.

Twenty-four saponins have been identified in alfalfa roots, including 13 medicagenic acids, 2 zanhic acids, 4 hederagenins, 1 soyasapogenol A, 2 soyasapogenol B's, 1 soyasapogenol E, and 1 bayogenin glycoside. Ten of the identified compounds, including 3-O-[beta-D-glucopyranosyl(1-->3)-beta-D-glucopyranosyl]-28-O-beta-D- glucopyranoside medicagenate, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta -D-glucopyranoside] medicagenic acid, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-beta- D-glucopyranoside medicagenate, 3-O-[beta-D-glucuronopyranosyl methyl ester]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1--> 2)-alpha-L-arabinopyranoside] medicagenate, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->2)-be ta-D-glucuronopyranosyl]-21-O-alpha-L-rhamnopyranoside soyasapogenol A, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)glucopy ranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl (1- ->2)-alpha-L-arabinopyranoside] medicagenate, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)glucopy ranosyl]-28-O-?beta-D-xylopyranosyl(1-->4)-)-[beta-D-apiofurano syl-(1 -->3)]- alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside? medicagenate, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyra nosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-O-?beta-D-xylopyranosyl(1-->4)-[beta-D-apiofurano side-(1-->3)]- alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside?zanhic acid, and 3-O-[beta-D-galactopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-28- O-b eta-D-glucopyranoside bayogenin, were not reported before, and their structures were established by spectral (FAB-MS and NMR) techniques. In addition, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->2)-be ta-D-glucuronopyranoside] soyasapogenol E was identified in the roots for the first time.     

Anthocyanins from new red leaf tea 'Sunrouge'

New red leaf tea cultivar 'Sunrouge' (Camellia taliensis × Camellia sinensis), for which an application for registration was made in 2009, is an anthocyanin-rich tea. The anthocyanin content of 'Sunrouge' was the highest among 4 tea cultivars, and was 8.4 times higher than that of 'Yabukita'. We purified and isolated 6 anthocyanins from 'Sunrouge' by chromatography, and identified them by LC/MS/MS and NMR analysis. As a result, the four anthocyanins were identified as delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (2), delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (3), cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (4), and cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (5), and the other two were estimated as delphinidin-(Z)-p-coumaroylgalactopyranoside (1), petunidin-(E)-p-coumaroylgalactopyranoside (6). Compound 3 was found in tea for the first time. In general, anthocyanins have various bioactivities, including relieving eyestrain and antioxidative effects, so it is expected that drinking 'Sunrouge' tea brings in similar bioactivities.     

外源硫酸铝调节八仙花花青苷组成和含量变化的分子生物学机制

【目的】八仙花是我国重要的观赏植物之一,本研究旨在探讨 Al₂(SO₄)₃ 对八仙花花色的影响及其机制。 【方法】以‘蓝色妈妈’品种为对象进行盆栽试验,设置了 0 (pH 为 6 的柠檬酸缓冲液)、2‰ 和 4‰ 3 个 Al₂(SO₄)₃ 水平,在植株出现花蕾约 1 cm 时进行处理。在开花盛期进行花色分析,采用高效液相色谱法和质谱分析花青苷成分和含量,采用电感耦合等离子体原子发射光谱法分析金属离子含量,采用荧光定量 PCR 分析 Al3+ 运输相关基因表达水平。 【结果】2‰ 和 4‰ 的 Al₂(SO₄)₃ 处理 21 d 后,花瓣颜色从粉色变为紫色和蓝紫色。‘蓝色妈妈’花瓣中检测到了飞燕草素-3-葡萄糖苷等 12 种花青苷;2‰ 和 4‰ 的 Al₂(SO₄)₃ 处理显著 (P < 0.05) 增加了飞燕草素苷、矢车菊素苷和芍药花素苷含量,其中增加幅度最大的是飞燕草素苷含量,从对照 (CK) 组的 5159.9 μg/g FW 分别增加到 24681.2 μg/g FW 和 30485.7 μg/g FW;飞燕草素苷含量增加主要是由于飞燕草素-3-葡萄糖苷和飞燕草素-3-戊糖-5-葡萄糖苷含量增加,飞燕草素-3-葡萄糖苷含量从对照的 4679.2 μg/g FW 分别增加到 23610.0 μg/g FW 和 29129.7 μg/g FW,飞燕草素-3-戊糖-5-葡萄糖苷从对照的 142.3 μg/g FW 分别增加到 805.6 μg/g FW 和 1114.9 μg/g FW。2‰ 和 4‰ 的 Al₂(SO₄)₃ 处理后,花瓣中 Al3+ 含量从对照的 2.24 μg/g FW 分别增加到 5.12 μg/g FW 和 11.83 μg/g FW;2‰ 和 4‰ 的 Al₂(SO₄)₃ 处理后,花瓣中质膜铝转运基因 (plasma membrane Al transporter, PALT) 和液泡膜铝转运基因 (vacuolar Al transporter, VALT) 表达水平显著提高 (P < 0.05),PALT 表达水平分别比对照提高了 88.5% 和 148.2%,VALT 表达水平分别比对照提高了 74.8% 和 135.7%。 【结论】Al₂(SO₄)₃ 处理诱导了 Al3+ 运输相关基因的表达,增加了花瓣中 Al3+ 积累,提高了飞燕草素苷含量,进而改变了花的颜色。     

Structures and antioxidant activity of anthocyanins in many accessions of eggplant and its related species

Anthocyanins were detected in extracts from the peels of 123 accessions of eggplant (Solanum melongena) and its related species. Their anthocyanin profiles were classified into four types, including known Japanese eggplant type (type 1) and non-Japanese eggplant type (type 2). Although most of the eggplant accessions had one of the two known profiles, one accession had a novel profile (type 3). Two accessions of related species showed another novel profile (type 4). The major anthocyanins were identified as delphinidin 3-(p-coumaroylrutinoside)-5-glucoside (nasunin) (type 1), delphinidin 3-rutinoside (type 2), delphinidin 3-glucoside (type 3), and petunidin 3-(p-coumaroylrutinoside)-5-glucoside (petunidin 3RGc5G) (type 4). Delphinidin 3-caffeoylrutinoside-5-glucoside (delphinidin 3RGcaf5G) was isolated from the hybrid (F1) plants of a type 1 cultivar and a type 3 germplasm. Among the five purified anthocyanins, delphinidin 3RGcaf5G showed the highest radical-scavenging activities toward both 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and linoleic acid radical, followed in order by nasunin and petunidin 3RGc5G.     

Isolation and identification of novel pyranoanthocyanins from black carrot (Daucus carota L.) juice

Six novel pyranoanthocyanins were identified by HPLC-ESI-MSn in black carrot (Daucus carota L. ssp. sativus var. atrorubens Alef.) juice. The two major compounds, namely, the vinylcatechol adducts of cyanidin 3-O-(6-O-feruloyl-beta-D-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside and cyanidin 3-O-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, respectively, were isolated by a combination of high-speed countercurrent chromatography with semipreparative HPLC. Their structures were fully elucidated by means of one- and two-dimensional NMR spectroscopy and high-resolution mass spectrometry. The four remaining pigments were characterized as the vinylphenol and vinylguaiacol adducts of cyanidin 3-O-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, the vinylguaiacol adduct of cyanidin 3-O-(6-O-feruloyl-beta-D-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, and the vinylcatechol adduct of cyanidin 3-O-(6-O-sinapoyl-beta-d-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside. These compounds are formed during storage of the juice through the direct reaction of either caffeic, ferulic, or coumaric acid with the respective genuine anthocyanins.     

Anthocyanins from black currants (Ribes nigrum L.)

Fifteen anthocyanin structures are reported from an extract of black currant berries (Ribes nigrum L.). These are the 3-O-glucosides and the 3-O-rutinosides of pelargonidin, cyanidin, peonidin, delphinidin, petunidin, and malvidin, cyanidin 3-O-arabinoside, and the 3-O-(6' '-p-coumaroylglucoside)s of cyanidin and delphinidin. The anthocyanins were characterized by means of size exclusion chromatography, high-performance liquid chromatography, UV-visible spectroscopy, and electrospray mass spectrometry. The four main pigments (the 3-O-glucosides and the 3-O-rutinosides of delphinidin and cyanidin) made up >97% of the total anthocyanin content. The minor pigments were enriched from the extract by successive partition against ethyl acetate and by gel fractionation. These chromatographic steps were successfully used to isolate the acylated anthocyanins from the ethyl acetate layer and to separate cyanidin 3-O-arabinoside from the mixture of anthocyanins. The amounts of anthocyanin rutinosides were found to be higher than the amount of the corresponding glucosides for all detected pigments having the same aglycon moiety.     

High-performance liquid chromatography with photodiode array detection (HPLC-DAD)/HPLC-mass spectrometry (MS) profiling of anthocyanins from Andean Mashua Tubers (Tropaeolum tuberosum Ruíz and Pavón) and their contribution to the overall antioxidant activity

Mashua (Tropaeolum tuberosum Ruíz and Pavón), an Andean tuber with high antioxidant activity, has sparked interest because of its traditional medicinal use. In this study, we evaluated the anthocyanin composition for three purple mashua genotypes and their contribution to the overall antioxidant activity of the tuber. Mashua anthocyanins, total phenolics, and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) antioxidant activity ranged from 45.5 to 131.9 mg of cyanidin 3-glucoside equivalents/100 g fresh weight (FW), 174.9 to 275.5 mg of gallic acid equivalents/100 g of FW, and 16.2 to 45.7 micromol of Trolox equivalents/g of FW, respectively. The high-performance liquid chromatography with photodiode array detection (HPLC-DAD) and HPLC-electrospray ionization tandem mass spectrometry (ESI/MS-MS) profiles revealed the presence of 11 different anthocyanins. The two major pigments (56.4-73.0% total area range at 520 nm) were identified as delphinidin 3-glucoside-5-acetylrhamnoside and delphinidin 3-sophoroside-5-acetylrhamnoside. Other pigments were delphinidin 3-glucoside-5-rhamnoside, delphinidin 3-sophoroside-5-rhamnoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, and cyanidin 3-sophoroside-5-rhamnoside. Cyanidin 3-glucoside and cyanidin 3-rutinoside were only found in two genotypes, while pelargonidin 3-sophoroside and pelargonidin 3-sophoroside-5-rhamnoside were only found in the third one. Anthocyanins from mashua were the major contributors to the total ABTS values for only one of the three genotypes, suggesting that other phenolics present are playing a major role in the antioxidant power of mashua tubers. Results from this study provide important information for the Nutraceutical and Functional Food Market for the use of mashua anthocyanins not only as a source of natural colorants but also as a source of phytonutrients.     

Reactivity of anthocyanins and pyranoanthocyanins. Studies on aromatic hydrogen-deuterium exchange reactions in methanol

Reactivity studies involving anthocyanin structures and their equilibrium forms will lead to better understanding of the properties of these antioxidants. Hydrogen-deuterium (H --> D) exchange reactions at various sites of the 3-glucosides of delphinidin (1), petunidin (2), malvidin (3), and the corresponding 3-glucosides of carboxypyranodelphinidin (4), carboxypyranopetunidin (5), carboxypyranomalvidin (6), and the flavonol quercetin 3-O-(6-alpha-rhamnopyranosyl-beta-glucopyranoside)(7) have been examined at room temperature in pure CD 3OD and in CD 3OD acidified with CF 3CO 2D. The H --> D exchange rate constants of H-6 and H-8 of 2 determined from (1)H NMR integration data were found to be independent upon pigment concentration (up to 4 x 10 (-2) M) and trifluoroactic acid concentration (0-15%, v/v), respectively. This suggest that these reactions follow first-order kinetics and unexpectedly to be independent of the acid concentration. H-6 and H-8 of the flavylium cation A-rings of 1- 3, and in the corresponding hydrogens of the hemiketal forms, exchanged with half-lives of approximately 100 h ( 1) and approximately 50 h ( 2 and 3), respectively. The pyranoanthocyanins (4-6) experienced no H --> D exchange for the analogous hydrogens, but H --> D exchange of H-beta (H-4)(t 1/2 approximately 25 h) for these compounds was observed. Only H-8 underwent significant H --> D exchange in 7. It is concluded that a stabilization of the sigma-complexes, assumed to be the intermediates in the reactions, takes place for the common anthocyanins (1-3) contrary to the pyranoanthocyanins (4-6).     

New oleanane saponins in Chenopodium quinoa

Six triterpenoid saponins were isolated from the seeds of Chenopodium quinoa (Chenopodiaceae). Their structures were as follows: phytolaccagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (1); spergulagenic acid 3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranoside (2); hederagenin 3-O-[beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (3); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (4); hederagenin 3-O-[beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (5); and spergulagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (6). Saponins 5 and 6 are new. The structures were characterized on the basis of hydrolysis and spectral evidence, including IR, UV, optical rotations, 1D- and 2D-NMR (HMQC and HMBC), ESIMS, and FABMS analyses.     

Structural identification of two major anthocyanin components of boysenberry by NMR spectroscopy

The major anthocyanins of boysenberry fruit, a cross between Rubus loganbaccus and Rubus baileyanus Britt., were isolated by preparative high-performance liquid chromatography (HPLC). The structures of cyanidin-3-[2-(glucosyl)glucoside] (1) and cyanidin-3-[2-(glucosyl)-6-(rhamnosyl)glucoside] (2) were determined by NMR in 1% DCOOD/D(2)O. An unusually high chemical shift (delta 2.5) is reported for H-5' ' of cyanidin-3-[2-(glucosyl)glucoside].     

Bioavailability and tissue distribution of anthocyanins in bilberry (Vaccinium myrtillus L.) extract in rats

To clarify how structural diversity of anthocyanins relates to their in vivo function, bioavailability was precisely studied in rats using bilberry (Vaccinium myrtillus L.) extract (Bilberon 25) as an anthocyanin source that contains 15 different anthocyanins. The bilberry extract was orally or intravenously administered to rats, and the plasma levels of each anthocyanin were determined by high-performance liquid chromatography. As the result, all anthocyanins except peonidin 3-O-alpha-L-arabinoside were detectable in the blood plasma. The plasma concentration of anthocyanins as a whole reached the maximum level of 1.2 microM at 15 min after oral administration of 400 mg/kg bilberry extract (153.2 mg/kg as anthocyanins) and then decreased with time. Uptake and decay profiles of each anthocyanin in the plasma were almost the same for all anthocyanins except a few with their maximum after 30 min. Among the anthocyanins carrying the same aglycone, the plasma level after 15 min of oral administration was as follows: galactoside > glucoside > arabinoside. Plasma clearance of anthocyanins after intravenous administration clearly showed that arabinoside disappeared more rapidly than glucoside and galactoside. On the other hand, when anthocyanins carrying the same sugar moiety were compared, the half disappearance time of plasma anthocyanins was in the following order: delphinidin > cyanidin > petunidin = peonidin > malvidin. The bioavailability of anthocyanins was in the range of 0.61-1.82% and was 0.93% as the anthocyanin mixture. The bioavailability of anthocyanins carrying the same aglycone was in the following order: Galactoside showed the highest followed by glucoside and arabinoside for cyanidin and delphinidin, but arabinoside and galactoside showed a higher bioavailability than glucoside for petunidin and malvidin. Anthocyanins recovered in urine and bile during the first 4 h after intravenous administration were only 30.8 and 13.4%, respectively. Anthocyanin profiles in tissues were quite different from those in blood plasma. The major anthocyanins distributed in liver and kidney were the O-methyl anthocyanins such as peonidin, malvidin, and other O-methyl anthocyanins derived from delphinidin, cyanidin, and petunidin-glycosides.     

Anthocyanin profile of Korean cultivated kidney bean (Phaseolus vulgaris L.)

This investigation was conducted to determine the structures and amounts of anthocyanins obtained from seed coats of kidney bean (Phaseolus vulgaris L.) cultivated in Korea. Anthocyanins in the seed coat of kidney bean were extracted with 1% HCl/20% CH(3)OH, and the crude anthocyanin extracts were purified by semipreparative HPLC. Five major anthocyanins were isolated, and their chemical structures were identified by spectroscopic methods (UV-vis, LC/ES-MS, and 1H and 13C NMR). The structures of these five anthocyanins were elucidated as cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-glucoside, petunidin 3-glucoside, and pelargonidin 3-glucoside. Using RP-HPLC with photodiode array detection, each of the five anthocyanins was separated within 12 min by using a gradient elution. It was proved that the application of RP-HPLC could be an excellent method for determining the composition and contents of anthocyanins in kidney bean. The preponderance of pelargonidin 3-glucoside and delphinidin 3-glucoside are observed in red and black kidney beans, respectively. However, in this study, it is reported for the first time that the contents and composition of anthocyanins in speckled seed depend on the classes of speckle color. The contents of cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-glucoside, petunidin 3-glucoside, pelargonidin 3-glucoside, and total anthocyanins in seed coats of 16 kidney beans cultivated in Korea were in the ranges of 0-0.04, 0-2.61, 0-0.12, 0-0.17, 0-0.59 and 0-2.78 mg/g of dried seed coats, respectively.     

Isolation and structural elucidation of two new glycosides from sage (Salvia officinalis L.)

Six compounds, 1-O-(2,3, 4-trihydroxy-3-methyl)butyl-6-O-feruloyl-beta-D-glucopyranoside, ethyl beta-D-glucopyranosyl tuberonate, p-hydroxybenzoic acid, (-)-hydroxyjasmonic acid, caffeic acid, and 4-hydroxyacetophenone 4-O-[5-O-(3, 5-dimethoxy-4-hydroxybenzoyl)-beta-D-apiofrunosyl]-(1-->2)-beta-D- glu copyranoside, were isolated from the n-butanol-soluble fraction of sage leaf extracts. Their structures were determined by spectral methods (MS, NMR, and 2D-NMR), and their antioxidant activities were measured. Among them, two new glycosides were elucidated. All of these compounds showed DPPH free radical scavenging activity at the concentration of 30 mM, and caffeic acid was the most active compound.     

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.     

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.     

New anthocyanidin and anthocyanin pigments from blue plumbago

Phytochemical investigations of blue plumbago ( Plumbago auriculata Poir. syn. Plumbago capensis Thunb.) flowers have led to the isolation of six new anthocyanins based on three new anthocyanidins with 5,7-dimethoxylated A-rings. Their structures were identified by 2D nuclear magnetic resonance and high-resolution mass spectrometry as the 3-O-β-galactopyranosides (1,2,4) and 3-O-α-rhamnopyranosides (3,5,6) of 5,7-dimethyldelphinidin, 5,7-dimethylpetunidin, and 5,7-dimethylmalvidin. Identification of 1-6 implies new structures for the previously reported anthocyanidins pulchellidin, europinidin, and capensinidin to be 5,7-dimethoxy-3,3',4',5'-tetrahydroxyflavylium, 5,7,3'-trimethoxy-3,4',5'-trihydroxyflavylium, and 5,7,3',5'-tetramethoxy-3,4'-dihydroxyflavylium cations, respectively. The anthocyanins (0.4 mg/g flowers) were accompanied by the dihydroflavonol taxifolin 3'-O-β-glucopyranoside (1.4 mg/g) and the flavonols 5-methylquercetin 3-O-α-rhamnopyranoside (8.8 mg/g) and 5-methylquercetin (0.4 mg/g). The anthocyanins 1-6 are the first reported natural anthocyanins with no free hydroxyl groups in their 5- and 7-positions on their A-rings. They have thus no possibility of forming the tautomeric quinonoidal bases (anhydrobases), which are related to the free hydroxyl groups in the 5- and 7-positions of previously reported anthocyanins. The genes behind the 5,7-dimethoxylated anthocyanins might be useful for making anthocyanins with special properties (colors, etc.).     

Triterpenoid glycosides from leaves of Medicago arborea L

Eighteen triterpene saponins (1-18) from Medicago arborea leaves have been isolated and their structures elucidated by spectroscopic, spectrometric (1D and 2D NMR, FAB-MS, ESI-MS/MS), and chemical methods. They have been identified as glycosides of medicagenic, zanhic, and 2beta-hydroxyoleanolic acids, soyasapogenol B, bayogenin, and 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid. Twelve of them, identified as 3-O-beta-D-glucopyranosyl-28-O-[alpha-L-arabinopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid (3), 3-O-beta-D-glucopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranosyl-(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid (4), 3-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranosyl(1-->2)-beta-D-glucopyranosyl]-2beta-hydroxyoleanolic acid (5), 3-O-beta-D-glucuronopyranosyl-28-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]medicagenic acid (6), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]bayogenin (9), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]-2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid (10), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (12), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranoside(1-->3)]-alpha-L-rhamnopyrano-syl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (13), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyrano-syl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (14), 3-O-[alpha-L-arabinopyranosyl-(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofurano-syl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (16), 3-O-[beta-D-glucopyrano-syl(1-->2)-beta-D-glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranosyl(1-->3)]-alpha-L-rhamno-pyranosyl (1-->2)-alpha-L-arabinopyranoside]zanhic acid (17), and 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyrano-side]medicagenic acid (18), are reported as new natural compounds. The presence of the aldehydic group on the sapogenin moiety of saponin 10 is discussed in the framework of a possible elucidation of the biosynthesis of these metabolites.     

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.