Structural determination of C-glycosylflavones by mass spectrometry of their permethyl ethers: O-glycosyl-6-C-glycosylflavones

Permethylated O-glycosyl-C-glycosylflavones give well defined MS including an important molecular peak. Permethyl 6-C-glycosylflavones O-glycosylated on a phenolic hydroxyl group are easily distinguished from the isomeric permethyl 6-C-diholosylflavones. In both types, the position of the O-glycosidic bond can be deduced from the MS, eventually after acid hydrolysis. 2″-O-glycosyl-6-C-glycosylflavones can be differentiated from their 8-C isomers.     

Sugar ring isomerization in C-arabinosylflavones

6-C-α-l-Arabinopyranosyl- and furanosylacacetins have been synthesized. They are isomerized by short acid treatment to give a mixture of the four anomer/ring size combinations without any Wessely-Moser isomerization. In the same conditions molludistin (8-C-α-l-arabinopyranosylgenkwanin) led only to a mixture of molludistin and 8-C-α-l-arabinofuranosylgenkwanin. This is the first demonstration of ring sugar isomerization in C-glycosylflavones. In usual solvent systems, α-anomers are easily separated from β-anomers, whereas corresponding pyranosyl and furanosyl anomers are not. However, they are easily separated after permethylation and characteristic features are found in the mass spectra of PM 6-C-arabinofuranosyl isomers.     

Fragmentation pattern of permethyl 6-C-glycosylflavones in electron impact mass spectrometry

A mass spectral fragmentation pattern of permethyl 6-C-glycosylflavones is proposed from the MS data of permethyl derivatives mono-O-deuteriomethylated in the 2″-, 3″-, 4″- or 6″-positions. The synthesis of these compounds via O″-glycosyl-6-C-glucosylflavones is described.     

Structural determination of 6-C-diglycosyl-8-C-glycosyl-flavones and 6-C-glycosyl-8-C-diglycosylflavones by mass spectrometry of their permethyl ethers

Permethylated 6-C-diglycosyl-8-C-glycosylflavones and 6-C-glycosyl-8-C-diglycosylflavones gave well defined EIMS including the molecular peak and a fragmentation pattern characteristic of the 6-C-glycosyl residue. X″′-O-glycosides (8-C-disaccharides) are thus easily distinguished from X″-O-glycosides (6-C-disaccharides) and, in the latter, the position of the O-glycosidic bond should be deduced from the MS, after acid hydrolysis. Three new C-glycosylflavones have been characterized in this way from Spergularia rubra and Stellaria holostea.     

C-glycosylanthocyanidins synthesized from C-glycosylflavones

Nine C-glycosyldeoxyanthocyanidins, 6-C-β-glucopyranosyl-7-O-methylapigeninidin, 6-C-β-glucopyranosyl-7-O-methylluteolinidin, 6-C-β-(2″-O-β-glucopyranosylglucopyranosyl)-7-O-methylapigeninidin, 6-C-β-(2″-O-β-glucopyranosylglucopyranosyl)-7,4′-di-O-methylapigeninidin, 8-C-β-glucopyranosylapigeninidin, 8-C-β-(2″-O-α-rhamnopyranosylglucopyranosyl)apigeninidin, 8-C-β-(2″-O-α-(4″′-O-acetylrhamnopyranosyl)glucopyranosyl)apigeninidin, 6,8-di-C-β-glucopyranosylapigeninidin (8), 6,8-di-C-β-glucopyranosyl-4′-O-methylluteolinidin (9), have been synthesized from their respective C-glycosylflavones (yields between 14% and 32%) by the Clemmensen reduction reaction using zinc-amalgam. The various precursors (C-glycosylflavones) of the C-glycosylanthocyanidins were isolated from either flowers of Iris sibirica L., leaves of Hawthorn ‘Crataegi Folium Cum Flore’, or lemons and oranges. This is the first time C-glycosylanthocyanidins have been synthesized. The structures of all flavonoids including the flavone rotamers were elucidated by 2D NMR techniques and high-resolution electrospray MS. The distribution of the various structural forms of 8 and 9 are different at pH 1.1, 4.5, and 7.0, however, the two pigments undergoes similar structural transformations at the various pH values. Pigments 8 and 9 with C-C linkages between the sugar moieties and the aglycone, were found to be far more stable towards acid hydrolysis than pelargonidin 3-O-glucoside, which has the typical anthocyanidin C-O linkage between the sugar and aglycone. This stability may extend the present use of anthocyanins as nutraceuticals, pharmaceuticals or colorants.     

Isocarlinoside,a di-C-glycosylflavone from Lespedeza capitata

Six di-C-glycosylflavones isolated from Lespedeza capitata leaves were identified as schaftoside, neoschaftoside, isoschaftoside, carlinoside, neocarlinoside and a new natural compound: isocarlinoside (6-C-α-l-arabinopyranosyl-8-C-β-d-glucopyranosylluteolin).     

The flavonoids of Trichophorum cespitosum

Fifteen flavonoids were isolated from Trichophorum cespitosum, including two new di-C-glycosylflavones, 6-C -arabinosyl-8-C-glucosylchr     

New C-glycosylflavones from Mollugo distica

Four C-glycosylflavones isolated from Mollugo distica were identified as 8-C-β-d-glucopyranosyl-genkwanin, 8-C-α-l-arabinopyranosylgenkwanin and their 2″-rhamnosides.     

Effect of conformation and configuration of arabinose residues on the circular dichroism of C-glycosylflavones

Examination of a variety of arabinose containing C-glycosylflavones has shown that the sign and intensity of the CD band at 250–275 nm (charge-transfer band) reflect not only the point of attachment of the sugar to the flavone but also depend upon the absolute and anomeric configuration, ring-size and ring-conformation in addition to the preferred rotameric conformation of the sugar about the C-aryl, C-l″ bond. A change in stereochemistry of arabinose from the α to β anomer resulted in sign inversion of the 250–275 nm CD band for 6-C-l-arabinosylflavones. Furthermore, a 6-C-arabinosylflavone containing α-l-arabinose exhibited an oppositely signed charge-transfer CD band in comparison to one which contained α-d-arabinose. 6,8-Di-C-glycosylflavones containing arabinose and glucose exhibited CD bands resulting from contributions due to both sugars, if the arabinose was not present as the β-pyranose form (¹C₄, conformation).     

Flavone di-C-glycosides from Scutellaria baicalensis

From the roots of Scutellaria baicalensis two new di-C-glycosylflavones have been isolated. Their structures have been established on the basis of mass, ¹H and ¹³C NMR spectroscopy as chrysin 6-C-glucoside-8-C-arabinoside and chrysin 6-C-arabinoside-8-C-glucoside.     

New C-glycosylflavones from Cerastium arvense

Three C-glycosylflavones isolated from Cerastium arvense have been identified as 6-C-xylosyl-apigenin (cerarvensin), its 7-O-glucoside and isovitexin 7,2″-di-O-glucoside.     

Antinociceptive activity of the HPLC- and MS-standardized hydroethanolic extract of Pterodon emarginatus Vogel leaves

Several studies have demonstrated the analgesic and anti-inflammatory effects of fruit and seed extracts from Pterodon emarginatus Vogel (Fabaceae). The objective of this study was to evaluate the antinociceptive activity of the hydroethanolic extract of P. emarginatus leaves in mice and characterize its chemical composition using HPLC coupled to UV–vis diode array detection and mass spectrometry with electrospray ionization. Our results showed that the doses of 500 and 1000 mg/kg produced an antinociceptive effect, as observed in the hot plate test and writhing induced by acetic acid. The chromatographic profile and spectral mass data suggest the presence of di-C-glycosylflavones (e.g., vicenin-2 and schaftoside), C,O-glycosylflavones (e.g., chrysoeriol-8-C-glucosyl-2″-O-glucuronide-6-C-arabinoside) and luteolin-7-O-rutinoside as the main constituents. Lower levels of oleanane-type saponins, such as soyasaponin Bb and Be, and the saponin derivatives hederagenin and aglycone B, which are typical of Fabaceae family, were also found. From this study, it is suggested that the analgesic effect observed is not due to the terpenoids previously reported from fruit and seed extracts, but could be attributed to flavones and the hederagenin derivatives that were identified as main constituents of the hydroethanolic extract from the leaves.     

Flavonoids and xanthones from the leaves of Amorphophallus titanum (Araceae)

The flavonoids and xanthones in the leaves of Amorphophallus titanum, which has the largest inflorescence among all Araceous species, were surveyed. Eight C-glycosylflavones, five flavonols, one flavone O-glycoside and two xanthones were isolated and characterized as vitexin, isovitexin, orientin, isoorientin, schaftoside, isoschaftoside, vicenin-2 and lucenin-2 (C-glycosylflavones), kaempferol 3-O-robinobioside, 3-O-rutinoside and 3-O-rhamnosylarabinoside, and quercetin 3-O-robinobioside and 3-O-rutinoside (flavonols), luteolin 7-O-glucoside (flavone), and mangiferin and isomangiferin (xanthones). Although the inflorescence of this species has been surveyed for flavonoids, those of the leaves were reported for the first time.     

Isomollupentin-O-glucosides from Cerastium arvense

Eight C-glycosylflavone O-glycosides including three new compounds: isomollupentin 7-O-glucoside, isomollupentin 4′-O-glucoside and isomollupentin 2″-O-glucoside have been isolated from the leaves and flowers of Cerastium arvense. The 27 C-glycosylflavones identified in this plant are tabulated.     

Flavonoid diversification in Achillea ptarmica and allied taxa

More than 50 collections of 12 species forming the A. ptarmica group have been analysed for their leaf flavonoids. C-Glycosylflavones (iso-orientin and derivatives, vicenins and lucenins) were found to be the main components, whereas flavonol 3-O-glycosides (based on quercetin and kaempferol) and flavone 7-O-glycosides (based on luteolin and diosmetin) were of restricted distribution. Infraspecific variability regarding C-glycosylflavones was observed in most of the taxa investigated. By contrast, flavonol 3-O-glycosides appeared to be stable characters and were sometimes accumulated instead of C-glycosylflavones. In addition to the flavonoids, the geographical distribution patterns and the possible origin of the A. sibirica in Eastern Asia are briefly discussed.     

Flavonoid pattern and systematics of the genus Leucocyclus

The flavonoid pattern of the monotypic Turkish genus Leucocyclus consists of C-glycosylflavones (isovitexin; isoorientin and derivatives; several di-C-glycosylapigenins; schaftoside, isoschaftoside and vicenin-3; lucenin-2), of flavonol 3-O-glycosides (quercetin and kaempferol 3-O-rhamnoglucoside) and trace amounts of luteolin 7-O-rhamnoglucoside. The systematic significance of the flavonoid diversification within Leucocyclus as well as possible relationships to other genera of the Anthemideae are discussed.     

The flavonoids of phragmites australis flowers

The major flavonoid constituents of Phragmites australis flowers are the C-glycosylflavones swertiajaponin, isoswertiajaponin and two new O-glycosides, the 3′-O-gentiobioside and the 3′-O-glucoside of swertiajaponin. Two unusual flavonol glycosides, rhamnetin 3-O-rutinoside and rhamnetin 3-O-glucoside, were also characterized from the same tissue.     

Flavonoids in the leaves of Hillebrandia and Begonia species (Begoniaceae)

The fresh leaves of Hillebrandia sandwicensis and 126 Begonia taxa were chemotaxonomically surveyed for flavonoids. Of their taxa, H. sandwicensis and 119 species, one variety and three hybrids were analyzed for flavonoids for the first time. Ten flavonols and eleven C-glycosylflavones were isolated and characterized as quercetin 3-O-rutinoside (1), kaempferol 3-O-rutinoside (2), isorhamnetin 3-O-rutinoside (3), quercetin 3-O-glucoside (4), quercetin 3-methyl ether 7-O-rhamnosylglucoside (5), quercetin 3,3'-dimethyl ether 7-O-rhamnosylglucoside (6), quercetin glycoside (13), quercetin glycoside (acylated) (14), kaempferol glycoside (17) and quercetin 3-O-rhamnoside (18) as flavonols, and isovitexin (7), vitexin (8), isoorientin (9), orientin (10), luteolin 6-C-pentoside (11), luteolin 8-C-pentoside (12), schaftoside (15), isoschaftoside (16), chrysoeriol 6,8-di-C-pentoside (19), apigenin 6,8-di-C-arabinoside (20) and isovitexin 2''-O-glucoside (21) as C-glycosylflavones. Quercetin 3-O-rutinoside (1) alone was isolated from H. sandwicensis endemic to Hawaii. Major flavonoids of almost Begonia species was also 1. Begonia species were divided into two chemotypes, i.e. flavonol containing type and C-glycosylflavone containing type. Of 14 section of the Begonia, almost species of many section, i.e. sect. Augustia, Coelocentrum, Doratometra, Leprosae, Loasibegonia, Monopteron and Ruizoperonia, were flavonol types. On the other hand, C-glycosyflavone type was comparatively most in sect. Platycentrum.     

C-glycosylflavones from roots of glycine max

Chemical investigation of the roots of Glycine max yielded six C-glycosylflavones identified on the basis of spectral data as carlinoside, isocarlinoside, vitexin, vitexin 2″-O-rhamnoside, isoschaftoside and a new compound 6,8-di-C-hexosylgenkwanin.     

Flavonoids in the leaves of twenty-eight polygonaceous plants

Flavonoids in the leaves of twenty-eight species belonging to the Polygonaceae were studied. Thirty-three kinds of flavonoids were isolated, and eighteen kinds were obtained as crystals. Quercetin glycosides were commonly found in the family. In the quercetin glycosides, 3-O-rhamnoside was most frequently found: 3-O-glucuronide is also distributed widely. Myricetin glycosides were rare. Methylated flavonols were found in some species of the sectionsEchinocaulon andPersicaria. Eleven kinds ofC-glycosylflavones were found in the present survey, andC-glycosylflavones were distributed in all species of the genusRheum and in almost all species of the section Tiniaria.Rumex Acetosella andPolygonum suffultum are exceptional, the former contains flavone glycoside and the latterC-glycosylflavones only, as main components.