Occurrence of steryl glycosides and acylated steryl glycosides in some marine algae

There is some controversy concerning the presence of steryl glycosides and acylated steryl glycosides in eucaryotic algae. These two classes of sterol compounds were investigated in species belonging to the three major groups of eucaryotic algae: green algae (Ulva gigantea, Cladophora rupestris), brown algae (Fucus vesiculosus, Ascophyllum nodosum), and red algae (Rhodymenia palmata, Porphyridium sp.). All these algae contain both steryl glycosides and acylated steryl glycosides. The sterol components of these compounds vary according to the alga but they are always the same as the free sterols of the alga in question. The most common sugar moiety is glucose. In the acylated steryl glycosides, the fatty acid is mainly palmitic acid. The percentage of these compounds (as a percentage of the total sterol content) is often low.     

Cardiac glycosides from Yellow Oleander (Thevetia peruviana) seeds

Thevetia cardiac glycosides can lead to intoxication, thus they are important indicators for forensic and pharmacologic surveys. Six thevetia cardiac glycosides, including two with unknown structures, were isolated from the seeds of the Yellow Oleander (Thevetia peruviana (Pers.) K. Shum., Apocynaceae). LC-ESI⁺–MS(/MS) analysis under high-resolution conditions used as a qualitative survey of the primary glycosides did not lead to fragmentation of the aglycones. Acid hydrolysis of the polar and non-volatile thevetia glycosides under severe conditions yielded the aglycones of the thevetia glycosides and made them amenable to GC–MS analysis. Comparison of mass spectral fragmentation patterns of the aglycones, as well as high-resolution mass spectrometric and NMR data of four of the primary thevetia glycosides including the two unknowns, revealed the structures of the complete set of six thevetia glycosides. The identified compounds are termed thevetin C and acetylthevetin C and differ by an 18,20-oxido-20,22-dihydro functionality from thevetin B and acetylthevetin B, respectively. The absence of an unsaturated lactone ring renders the glycosides cardio-inactive. The procedures developed in this study and the sets of analytical data obtained will be useful for screening and structure assessment of other, particularly polar, cardiac glycosides.     

Secoiridoid glycosides production by Centaurium maritimum (L.) Fritch hairy root cultures in temporary immersion bioreactor

Secoiridoid glycosides are naturally occurring phytochemicals of great importance for the food and pharmaceutical industry because of their various biological activities. Certain Gentiana and Centaurium species, which are recognized as the most important sources of these compounds, have become critically endangered due to overexploitation. In this study we describe a laboratory-scale approach for further implementation in large-scale production of secoiridoid glycosides, using a hairy root culture system of Centaurium maritimum L. Fritch, an underutilized and phytochemically unexplored species. Hairy roots were induced by Agrobacterium rhizogenes strain A40M70GUS and grown in Erlenmeyer flasks, as well as in RITA^® temporary immersion bioreactors (TIBs). About 2–4 times higher biomass production rate and up to 8 times higher total secoiridoid glycosides production rate were achieved in RITA^® bioreactors. Among the selected hairy root lines, line HR3 cultured in RITA^® TIBs proved to be the most efficient considering both biomass and secoiridoid glycosides production rate.     

Identification of cardiac glycosides in fractions from Periploca forrestii by high-performance liquid chromatography/diode-array detection/electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/nuclear magnetic resonance

Cardiac glycosides are a class of naturally occurring compounds that are characterized by some interesting biological activities and are widely distributed in the plant kingdom and can also be found in some animals. There is an interest in the chemical characterization of these molecules due to their toxicity and their use in medicines. In the study reported here, a combination of electrospray ionization tandem mass spectrometry with high-performance liquid chromatography equipped with diode-array detector (HPLC-DAD/ESI-MSⁿ), and hyphenation to both liquid chromatography and nuclear magnetic resonance spectroscopy (HPLC/NMR) were utilized for the on-line analyses of cardiac glycosides from Periploca forrestii. The fragmentation patterns and ¹H NMR spectra of nine isolated cardiac glycosides were investigated; their fragmentation rules and ¹H NMR spectral characteristics were summarized and applied to the structural identification of similar constituents in fractions from P. forrestii. As a result, a total of nine trace cardiac glycosides were tentatively determined by analyses of accurate molecular masses, representative fragment ions and characteristic ¹H NMR signals provided by HPLC/high-resolution mass spectrometry (HRMS), HPLC-DAD/ESI-MSⁿ and HPLC/¹H NMR experiments, respectively. Of these, eight (2–9) are new compounds and one (1) is reported from P. forrestii for the first time. Results of the present study can benefit the rapid identification and targeted isolation of new cardiac glycosides from crude plant extracts.     

Changes in citrus leaf flavonoid concentrations resulting from blight-induced zinc-deficiency

Increased flavonoid concentrations were found to correlate with the elevated levels of leaf phenolic compounds occurring in blight-induced zinc-deficient citrus. In orange (Citrus sinensis L.) leaves, the increases occurred primarily in hesperidin and diosmin, whereas in grapefruit (C. paradisi Macf.) the largest increases occurred in naringin and rhoifolin. Zinc-deficiency occurring in the blighted citrus leaves appeared to be the important contributing factor to the increased flavonoid content. Although the leaves from trees with blight were typically smaller than leaves from unaffected trees, the increased flavonoid content was not significantly due to a concentration effect. Large differences occurred in the percent increases in concentrations of certain citrus leaf flavonoids. While large increases occurred for a number of flavanone and flavone glycosides, much smaller percent increases occurred for other minor flavone glycosides, and the polymethoxyflavone aglycones. The parallel increases occurring in the concentrations of certain flavone glycosides and their flavanone analogs provide a further indication that flavanone glycosides are precursors in the biosynthesis of flavone glycosides in citrus.     

Cardiotonic glycosides from the latex of naucleopsis mello-barretoi,a dart-poison plant from North-West Brazil

Poison scraped from blowpipe darts made by the Makti Indians of the Upper Rio Uneiuxi, Rio Negro basin, and prepared from the latex of Naucleopsis mello-barretoi (Standl.) C. C. Berg, contained ca. 4.4% cardiotonic glycosides; one of the major glycosides present wasα-antiarin. Latex fromN. mello-barretoi yielded ca. 1.3% of a complex mixture of cardiotonic glycosides; the major component was β-antiarin and it was accompanied by small amounts of α-antiarin, probably antioside, and unidentified glycosides. The bark of the plant had only traces of cardiotonic glycosides; some components appeared to be the same as those in the latex, but the antiarins and antioside were absent. Cardiotonic glycosides were not detected in the leaves.     

Diosgenin saponins from Dioscorea floribunda

Five spirostanol glycosides and two furostanol glycosides were isolated from Dioscorea floribunda. In addition to the IR spectra of the free glycosides and the MS of the peracetates and permethyl ethers, the most effective method for structural determination proved to be the NMR spectra of the free saponins in pyridine-d₅.     

Flavonoid glycosides of Tribulus pentandrus and T. Terrestris

Twenty-five flavonoid glycosides were detected in Tribulus pentandrus and T. terrestris. The glycosides belong to the common flavonols, kaempferol, quercetin and isorhamnetin, with the 3-gentiobiosides as the major glycosides. Traces of a flavone (tricin) glycoside was also present in T. pentandrus. The separation of Tribulaceae as a distinct family from Zygophyllaceae is discussed.     

Flavones and anthocyanins from the leaves and flowers of Japanese Ajuga species (Lamiaceae)

Flavones and anthocyanins were isolated from the leaves and flowers of 14 Ajuga taxa (Lamiaceae), which are all native or naturalized in Japan. Of 13 flavones obtained from the leaves, 11 were characterized as apigenin, luteolin, 6-hydroxyluteolin and acacetin glycosides. Ten flavones were isolated from the flowers. Ten anthocyanins were isolated from the flowers. Six of these anthocyanins were identified as acylated delphinidin glycosides and four were shown to be acylated cyanidin glycosides. Japanese Ajuga taxa were chemotaxonomically discussed by their distribution patterns, especially foliar flavonoids.     

Impact of the dual defence system of Plantago lanceolata (Plantaginaceae) on performance,nutrient utilisation and feeding choice behaviour of Amata mogadorensis larvae (Lepidoptera,Erebidae)

Iridoid glycosides are plant defence compounds with potentially detrimental effects on non-adapted herbivores. Some plant species possess β-glucosidases that hydrolyse iridoid glycosides and thereby release protein-denaturing aglycones. To test the hypothesis that iridoid glycosides and plant β-glucosidases form a dual defence system, we used Plantago lanceolata and a polyphagous caterpillar species. To analyse the impact of leaf-age dependent differences in iridoid glycoside concentrations and β-glucosidase activities on insect performance, old or young leaves were freeze-dried and incorporated into artificial diets or were provided freshly to the larvae. We determined larval consumption rates and the amounts of assimilated nitrogen. Furthermore, we quantified β-glucosidase activities in artificial diets and fresh leaves and the amount of iridoid glycosides that larvae feeding on fresh leaves ingested and excreted. Compared to fresh leaves, caterpillars grew faster on artificial diets, on which larval weight gain correlated positively to the absorbed amount of nitrogen. When feeding fresh young leaves, larvae even lost weight and excreted only minute proportions of the ingested iridoid glycosides intact with the faeces, indicating that the hydrolysis of these compounds might have interfered with nitrogen assimilation and impaired larval growth. To disentangle physiological effects from deterrent effects of iridoid glycosides, we performed dual choice feeding assays. Young leaves, their methanolic extracts and pure catalpol reduced larval feeding in comparison to the respective controls, while aucubin had no effect on larval consumption. We conclude that the dual defence system of P. lanceolata consisting of iridoid glycosides and β-glucosidases interferes with the nutrient utilisation via the hydrolysis of iridoid glycosides and also mediates larval feeding behaviour in a concentration- and substance-specific manner.     

Flavonol glycosides of Euphorbia retusa and E. sanctae-catharinae

The flavonoid glycosides of Euphorbia retusa and E. sanctae-catharinae are reported. Besides a number of common flavonol glycosides, kaempferol and quercetin 3-glucuronide-7-glucosides are reported for the first time.     

A chemotaxonomic study of flavonoids from european teucrium species

A survey of aerial tissues of 42 European taxa of the genus Teucrium has revealed the widespread presence of five surface flavonoids: cirsiliol, cirsimaritin, cirsilineol, salvigenin and 5-hydroxy-6,7,3′,4′-tetramethoxyflavone. The latter two compounds are useful taxonomic markers in that salvigenin is characteristic of species of section Polium, while 5-hydroxy-6,7,3′,4′-tetramethoxyflavone is completely confined to species of the other five sections surveyed. Eleven flavone glycosides, four flavonol glycosides and the glycoflavone vicenin-2 were found to occur as vacuolar constituents. One of the flavone glycosides, cirsimaritin 4′-glucoside, only occurs in the species T. arduini, while two others, hypolaetin and isoscutellarein 7-acetyl-allosylglucosides, are characteristic of the closely related T. chamaedrys and T. webbianum. 6-Hydroxyluteolin is widely present as the 7-glucoside and 7-rhamnoside, the latter compound being a new glycoside. In general, the chemical results are correlated with sectional classification and usefully indicate that at least one taxon, T. compactum, is misplaced within the genus. Phyletically, the restriction of flavonol glycosides mainly to section Teucrium suggests that this may be the basic group within the genus.     

Potential benefits of iridoid glycoside sequestration in Longitarsus melanocephalus (Coleoptera,Chrysomelidae)

Whenever potentially noxious plant compounds are taken up and recycled by herbivorous insects, a protective function of these sequestered compounds is assumed. The flea beetle Longitarsus melanocephalus sequesters iridoid glycosides from its host plant up to a concentration of 2% DW, yet so far it remained unknown whether the insects gain protection from natural enemies by sequestering plant compounds at these comparatively low concentrations. Here we tested whether iridoid glycosides might deter or inhibit various soil and litter-dwelling potential enemies and pathogens. In choice experiments presenting L. melanocephalus pupae together with Tribolium castaneum pupae, the predator Lithobius forficatus was deterred by the iridoid glycoside containing pupae, while Forficula auricularia as well as the nematode Heterorhabditis bacteriophora were not deterred. L. forficatus also avoided artificial baits doted with 2% iridoid glycosides while F. auricularia showed no aversion to iridoid glycosides at these concentrations and H. bacteriophora did not suffer any toxic effect. Of the pathogens tested, the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae were not inhibited in their growth by iridoid glycosides ranging up to 2%. However, an inhibitory effect could be observed against the entomopathogenic bacterium Bacillus thuringiensis, even at the relatively small concentrations that are common in L. melanocephalus. The antibacterial effect might thus be another important selective value of iridoid glycoside sequestration in this species.     

Flavonoid glycosides of Lathyrus pratensisleguminosae

Eight flavonoid glycosides have been identified in the leaves of Lathyrus pratensis, two of which are new glycosides.     

La systématique infraspécifiquede l'Anthyllis vulneraria (Leguminosae) vue à la lumière de la biochimie flavonique

Twenty-six experimental populations of A. vulneraria were surveyed for leaf flavonoids. The distribution of 34 flavonol glycosides showed, after factor analysis, the existence of for distinct chemical taxa, each exhibiting characteristic glycosides of 7-methylkaempferol. These results, and others obtained from further factor analysis, are discussed in relation to Couderc's taxonomic studies, and allow a better definition of infraspecific units of A. vulneraria. Flavonoid and morphological data are generally correlated.     

Molecular design of N-linked tetravalent glycosides bearing N-acetylglucosamine,N,N′-diacetylchitobiose and N-acetyllactosamine: Analysis of cross-linking activities with WGA and ECA lectins

Two types of nonspacer- and spacer-N-linked tetravalent glycosides bearing N-acetylglucosamine (GlcNAc), N,N′-diacetylchitobiose [(GlcNAc)₂] and N-acetyllactosamine (LacNAc) were designed and prepared as glycomimetics. The interactions of wheat germ (Triticum vulgaris) agglutinin (WGA) and coral tree (Erythrina cristagalli) agglutinin (ECA) with a series of tetravalent glycosides and related compounds were studied using a hemagglutination inhibition assay, a precipitation assay, double-diffusion test, and an optical biosensor based on surface plasmon resonance (SPR). The tetravalent glycosides were found to be capable of binding and precipitating the lectins as tetravalent ligands. Strong interactions with WGA, due to a combination of multivalency effects and spacer effects, were observed for tetravalent glycosides bearing flexible tandem GlcNAc. The chelate effect leads to large rate enhancement for the tetravalent system with favorable orientation of ligands. Our simple strategy produced multivalent glycosides with strong cross-linking activity for lectin as a specific coagulant.     

Preparation of 6-aminohexyl d-aldopyranosides

6-Aminohexyl glycosides covalently linked to solid matrices are effective reagents for the isolation of proteins that bind to carbohydrates [Schnaar and Lee, Biochemistry, 14 (1975) 1535–1541], and for the study of interactions between intact cells and immobilized carbohydrates [Weigel et al., J. Biol. Chem., 253 (1978) 330–333]. The preparation of the 6-aminohexyl glycosides of the following D-pyranoses is now described: β-glucose, β-galactose, 2-acetamido-2-deoxy-β-glucose, α-mannose, β-maltose, β-melibiose, β-lactose, and β-cellobiose. These glycosides were prepared by glycosylation of 6-(trifluoroacetamido)hexanol with the appropriate acetylated glycosyl halide in 1:1 (v/v) benzene-nitromethane, with mercuric cyanide as the catalyst. Deacylation of the glycosides was achieved in two steps: use of sodium methoxide for O-deacetylation, and of an anion-exchange resin for N-de(trifluoroacetyl)ation.     

Veronica: Acylated flavone glycosides as chemosystematic markers

HPLC/DAD and LC–MS of an extract of Veronica spicata (subgenus Pseudolysimachium, Plantaginaceae) revealed the presence of six 6-hydroxyluteolin glycosides acylated with phenolic acids, three of which are new compounds and which we called spicosides. A flavonoid survey of seven more species belonging to V. subgenus Pseudolysimachium and eight species of V. subgenus Pentasepalae showed that all the Pseudolysimachium species and four of the Pentasepalae species produced 6-hydroxyflavone glycosides, whereas the remaining four Pentasepalae species contained acetylated 8-hydroxyflavone glycosides instead. Spicosides appeared to be common in subgenus Pseudolysimachium (detected in five out of eight species), but we did not find them in subgenus Pentasepalae. Previously, acetylated 8-hydroxyflavone glycosides have been isolated from or detected in eight species of V. subgenus Pentasepalae (in 13 species altogether including the new results) and seven species of V. subgenus Pocilla. However, 6-hydroxyflavone glycosides occur in some other species of these subgenera. The results are discussed in an evolutionary context.     

The occurrence of acylated flavonol glycosides in the cruciferae

Nine acylated glycosides of kaempferol or quercetin were identified in Sisymbrium gilliesii, and in three Crambe spp. They were usually present together with the related unacylated glycosides. Acylation is a very common characteristic of the four crucifer species studied.     

Effect of chito-oligosaccharide on the intestinal absorptions of phenylethanoid glycosides in Fructus Forsythiae extract

Phenylethanoid glycosides, the main active ingredients in Fructus Forsythiae extract possesses strong antibacterial, antioxidant and antiviral effects, and their contents were higher largely than that of other ingredients such as lignans and flavones, but their absolute bioavailability orally was significantly low, which influenced clinical efficacies of its oral preparations seriously. In the present study, the absorption mechanism of phenylethanoid glycosides was studied using in vitro Caco-2 cell model. And the effect of chito-oligosaccharide (COS) on the intestinal absorption of phenylethanoid glycosides in Fructus Forsythiae extract was investigated using in vitro, in situ and in vivo models. The pharmacological effects such as antiviral activity improvement by COS were verified by MDCK cell damage inhibition rate after influenza virus propagation. The observations from in vitro Caco-2 cell showed that the absorption of phenylethanoid glycosides in Fructus Forsythiae extract so with that in monomers was mainly restricted by the tight junctions, and influenced by efflux transporters (P-gp and MRP2). Meanwhile, the absorption of phenylethanoid glycosides in Fructus Forsythiae extract could be improved by COS. Besides, COS at the same low, medium and high concentrations caused a significant, concentration-dependent increase in the P_app-value for phenylethanoid glycosides compared to the control group (p < 0.05), and was all safe for the Caco-2 cells. The observations from single-pass intestinal perfusion in situ model showed that the intestinal absorption of phenylethanoid glycosides can be enhanced by COS. Meanwhile, the absorption enhancing effect of phenylethanoid glycosides might be saturable in different intestine sites. In pharmacokinetics study, COS at dosage of 25 mg/kg improved the bioavailability of phenylethanoid glycosides in Fructus Forsythiae extract to the greatest extent, and was safe for gastrointestine from morphological observation. In addition, treatment with Fructus Forsythiae extract with COS at dosage of 25 mg/kg prevented MDCK cell damage upon influenza virus propagation better than that of control. All findings above suggested that COS at dosage of 25 mg/kg might be safe and effective absorption enhancer for improving the bioavailability of phenylethanoid glycosides and the antiviral activity in vitro in Fructus Forsythiae extract.