UPLC-MS/MS Profile of Alkaloids with Cytotoxic Properties of Selected Medicinal Plants of the Berberidaceae

Cancer is one of the most occurring diseases in developed and developing countries. Plant-based compounds are still researched for their anticancer activity and for their quantity in plants. Therefore, the modern chromatographic methods are applied to quantify them in plants, for example, UPLC-MS/MS (ultraperformance liquid chromatography tandem mass spectrometry). Therefore, the aim of the present study was to evaluate the content of sanguinarine, berberine, protopine, and chelidonine in Dicentra spectabilis (L.) Lem., Fumaria officinalis L., Glaucium flavum Crantz, Corydalis cava L., Berberis thunbergii DC., Meconopsis cambrica (L.) Vig., Mahonia aquifolium (Pursh) Nutt., Macleaya cordata Willd., and Chelidonium majus L. For the first time, N,N-dimethyl-hernovine was identified in M. cambrica, B. thunbergii, M. aquifolium, C. cava, G. flavum, and C. majus; methyl-hernovine was identified in G. flavum; columbamine was identified in B. thunbergii; and methyl-corypalmine, chelidonine, and sanguinarine were identified in F. officinalis L. The richest source of protopine among all the examined species was M. cordata (5463.64 ± 26.3 μg/g). The highest amounts of chelidonine and sanguinarine were found in C. majus (51,040.0 ± 1.8 μg/g and 7925.8 ± 3.3 μg/g, resp.), while B. thunbergi contained the highest amount of berberine (6358.4 ± 4.2 μg/g)

Phenolics in aerial parts of Persian clover Trifolium resupinatum

The nutritional quality of Persian clover (Trifolium resupinatum), an important pasture crop, depends not only on a high protein content but also on the occurrence of animal health and welfare promoting phytochemicals. Nine phenolic constituents present in the aerial parts of this species were isolated and their structures confirmed by NMR and ESI-MS analyses. The compounds included two chlorogenic acids, four quercetin and two kaempferol glycosides, as well as the isoflavone formononetin-7-glucoside. The concentration of isoflavone was low, not exceeding 1.2 mg/g of dry matter. The concentration of flavonols ranged between 5.9 and 11.8 mg/g, depending on the sampling dates, with the highest concentration occurring in the first cut. A similar trend in the concentration was found for chlorogenic acids, which ranged from 2 mg/g in summer to 7.3 mg/g in spring

The anti-adhesive and anti-aggregatory effects of phenolics from Trifolium species in vitro

Abstract The present in vitro study includes a compar-ative evaluation of anti-platelet (anti-thrombotic) proper-ties of plant phenolics, isolated from nine different clover (Trifolium) species. The analysis covered phenolic frac-tions isolated from T. alexandrinum L., T. fragiferum L., T. hybridum L., T. incarnatum L., T. pallidum Waldst et Kit., T. resupinatum L. var. majus Boiss, T. resupinatum L. var. resupinatum, T. scabrum L., and T. pratense L. (red clo-ver). The inhibitory effects of plant preparations (1–50 lg/ ml) on hemostatic functions of blood platelets were assessed by measurements of thrombin- or ADP-induced platelet adhesion to fibrinogen, platelet aggregation in platelet-rich plasma (activated with ADP or collagen), and by the determination of PF-4 secretion from platelet a-granules. The influence of T. phenolics on arachidonic cascade in blood platelets was also determined. T. resupinatum var. majus, T. resupinatum var. resupinatum, and T. scabrum had the strongest anti-platelet effects. These preparations displayed the most evident anti-adhe-sive and anti-aggregatory effects in response to all of the used agonists: thrombin (0.2 U/ml), ADP (10 lM), and collagen (2 lg/ml), and their inhibitory properties were also confirmed by an analysis of PF-4 secretion. T. scabrum and some of other examined clover species possess sig-nificantly higher concentrations of both isoflavones and other bioactive phenolics, when compared to red clover. The obtained results suggest that these clovers contain substances with potent anti-platelet properties

Assessment of allelopathic potential of Solidago gigantea Aiton on dry weight of Echinochloa crus-galli (L.) Beauv. and Amaranthus retroflexus L.

Laboratory analyses using the 1st generation bioassay were conducted in the years 2013-2014 to investigate the allelopathic potential of wateralcoholic and aqueous extracts from dry weight of rhizomes and roots as well as stems and leaves of Solidago gigantea. Analysed acceptors were two weed species, i.e. monocotyledonous Echinochloa crus-galli and dicotyledonous Amaranthus retroflexus. When the acceptors (E. crus-galli and A. retroflexus) reached the 2-leaf stage (BBCH 12) they were sprayed with wateralcoholic and aqueous extracts (at concentrations of 12.5%, 10%, 5% and 2.5%) obtained from the donor, i.e. S. gigantea. Results indicate an inhibitory effect of wateralcoholic extracts from aboveground parts (leaves and stems) of S. gigantea in relation to dry weight of E. crus-galli and A. retroflexus. The volume of dry weight reduction in acceptors was dependent on the concentration of extracts produced from the donor plant S. gigantea. Dry weight of E. crus-galli and A. retroflexus was reduced most effectively by two concentrations: 12.5% and 10%. In turn, aqueous extracts from rhizomes and roots of S. gigantea, irrespective of the applied concentration, caused an increase in dry weight of E. crus-galli and A. retroflexus. Only aqueous extracts produced from leaves and stems of S. gigantea, irrespective of their concentration, reduced dry weight in only E. crus-galli

Variation in Flavonoids in Leaves, Stems and Flowers of White Clover Cultivars:

In the present study, the major flavonoids of white clover ( Trifolium repens L.) cv. Sonja were extracted, isolated and identified. The major flavonoids in leaves and stems were the four flavonol glycosides: kaempferol-3- O-{Xyl(1→2)-Gal} (kaempferol-Xyl-Gal), kaempferol-3- O-{Rha(1→6)-[Xyl(1→2)]-Gal} (kaempferol-Rha-Xyl-Gal), quercetin-3- O-{Xyl(1→2)-Gal} (quercetin-Xyl-Gal), and quercetin-3- O-{Rha(1→6)-[Xyl(1→2)]-Gal} (quercetin-Rha-Xyl-Gal). Quercetin-Rha-Xyl-Gal has never been reported before and kaempferol-Rha-Xyl-Gal has not previously been identified in clover aerial parts. Concentrations of those compounds, together with aglyconic flavonoids previously described in white clover, as well as their glycosides, were quantified in leaves/stems and flowers of four white clover cvs Rabani, Klondike, Ramona and Aran using tandem mass spectrometry. There were significant differences in flavonoid concentrations in the two plant parts, with the highest concentrations of most aglycones in flowers and the highest concentrations of most glycosides in leaves/stems. This distribution of compounds may indicate different ways of storage and/or different mechanisms of action of the compounds. The cultivars were selected for genetic diversity, which resulted in distinctly different amounts of flavonoids in the plants. Concentrations of 17 of 24 compounds varied significantly – for some compounds up to a factor of 10 – among cultivars. Total flavonoid concentrations in flowers did not vary greatly among cultivars, at 28.9–35.8 mmol/g dry material (DM). In contrast, in leaves/stems, the cvs Rabani and Klondike had lower concentrations of most flavonoids (total concentrations 10.0 and 12.7 mmol/gDM, respectively) compared to cvs Aran and Ramona (32.3 and 22.1 mmol/gDM, respectively). There is a potential for breeding/selection of cultivars with targeted concentrations of particular flavonoids

Effects of two sources of tannins (Quercus L. and Vaccinium vitis idaea L.) on rumen microbial fermentation: an in vitro study

The aim of the experiment was to determine the effect of different sources of tannins on the in vitro rumen fermentation with focus on methane production. In the experiment, a rumen simulation system (RUSITEC) equipped with 4 fermenters (1 L) was used in three replicated runs (6 d of adaptation and 4 d of sampling) to study the effects of Quercus cortex extract (QC), Vaccinium vitis idaea (VVI) dried leaf extract and a mixture of VVI/QC on rumen microbial fermentation. Fermenters were fed 10.9 g/d of dry matter (DM) of a 600:400 forage:concentrate diet. Treatments were control, QC (2.725 mL), VVI leaves 0.080 g) and mixture of QC/VVI (1.362 mL+0.040 g) and were randomly assigned to fermenters within periods. The equivalent of 2.5 g of tannins/kg dietary DM from three sources of tannins was evaluated. All tannin sources decreased CH4 and ammonia concentrations, as well as protozoa and methanogen counts (P<0.001). Vaccinium vitis idaea and QC/VVI tended (P=0.005) to reduce the acetate to propionate ratio. There were no changes in nutrient digestion. Results suggest that these sources of tannins, especially VVI have the potential to reduce rumen CH4 production and ammonia concentration without negative effects on in vitro DM digestibility, total volatile fatty acids and pH

Isolation and Structural Determination of Triterpenoid Glycosides from the Aerial Parts of Alsike Clover (Trifolium hybridum L.)

Five azukisapogenol glycosides (1−5) have been isolated from the aerial parts of alsike clover (Trifolium hybridum L.), and their structures were elucidated by combined spectroscopic, spectrometric (1D and 2D NMR; HRESIMS, ESI−MS/MS), and chemical methods. Three of them are new compounds and were identified as 3-O-[-α-L-arabinopyranosyl(1→2)]-β-Dglucuronopyranosyl azukisapogenol (1), 3-O-[-β-D-glucuronopyranosyl(1→2)-β-D-glucuronopyranosyl]-29-O-β-D-glucopyranosyl azukisapogenol (2), and 3-O-[-α-L-arabinopyranosyl(1→2)-β-D-glucuronopyranosyl]-29-O-β-D-glucopyranosyl azukisapogenol (3). The remaining two (4, 5) are known compounds but have not been previously described as saponins constituents of the genus Trifolium. Also, azukisapogenol is reported here as a triterpenoid aglycone for the first time in this genus. Finally, the main chemotaxonomic features that may be recognized as specific of Trifolium species were discussed

Bioactive steroidal saponins from Agave offoyana flowers

Bioguided studies of flowers of Agave offoyana allowed the isolation of five steroidal saponins never described previously, Magueyosides A–E (1–5), along with six known steroidal saponins (6–11). The structures of compounds were determined as (25R)-spirost-5-en-2a,3b-diol-12-one 3-O-{b-D-xylopyranosyl-(1-3)-O-b-D-glucopyranosyl-(1-2)-O-[b-D-xylopyranosyl-(1-3)]-O-b-D-glucopyranosyl-( 1-4)-O-b-D-galactopyranoside} (1), (25R)-spirost-5-en-2a,3b-diol-12-one 3-O-{b-D-glucopyranosyl-(1-2)-O-[b-D-xylopyranosyl-(1-3)]-O-b-D-glucopyranosyl-(1-4)-O-b-D galactopyranoside} (2), (25R)-spirost-5-en-2a,3b,12b-triol 3-O-{b-D-glucopyranosyl-(1-2)-O-[b-D-xylopyranosyl-(1-3)]- O-b-D-glucopyranosyl-(1-4)-O-b-D-galactopyranoside} (3), (25R)-5a-spirostan-2a,3b-diol-12-one 3-O-{b-D-xylopyranosyl-(1-3)-O-b-D-glucopyranosyl-(1-2)-O-[b-D-xylopyranosyl-(1-3)]-O-b-D-glucopyranosyl-(1-4)-O-b-D-galactopyranoside} (4), and (25R)-5a-spirostan-2a,3b-diol-9(11)-en-12-one 3-O-{b-D-xylopyranosyl-(1-3)-O-b-D-glucopyranosyl-(1-2)-O-[b-D-xylopyranosyl-(1-3)]-O-b-D-glucopyranosyl-( 1-4)-O-b-D-galactopyranoside} (5), by comprehensive spectroscopic analysis, including one- and two-dimensional NMR techniques, mass spectrometry and chemical methods. The bioactivities of the isolated compounds on the standard target species Lactuca sativa were evaluated. A dosedependent phytotoxicity and low dose stimulation were observed

Phytotoxic steroidal saponins from Agave offoyana leaves

A bioassay-guided fractionation of Agave offoyana leaves led to the isolation of five steroidal saponins (1–5) along with six known saponins (6–11). The compounds were identified as (25R)-spirost-5-en-2α,3β-diol-12-one 3-O-{α-L-rhamnopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-O-β-D-glucopyranosyl-(1→4)-O-β-D-galactopyranoside} (1), (25R)-spirost-5-en-3β-ol-12-one 3-O-{α-L-rhamnopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-O-β-D-glu copyranosyl-(1→4)-O-β-D-galactopyranoside} (2), (25R)-spirost-5-en-3β-ol-12-one 3-O-{β-D-xylopyrano syl-(1→3)-O-β-D-glucopyranosyl-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-O-β-D-glucopyranosyl-(1→4)-O-β -D-galactopyranoside} (3), (25R)-26-O-β-D-glucopyranosylfurost-5-en-3β,22α,26-triol-12-one 3-O- {α-L-rhamnopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-O-β-D-glucopyrano syl-(1→4)-O-β-D-galactopyranoside} (4) and (25R)-26-O-β-D-glucopyranosylfurost-5-en-3β,22α,26-triol- 12-one 3-O-{β-D-xylopyranosyl-(1→3)-O-β-D-glucopyranosyl-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-O-β- D-glucopyranosyl-(1→4)-O-β-D-galactopyranoside} (5) by comprehensive spectroscopic analysis, including one- and two-dimensional NMR techniques, mass spectrometry and chemical methods. The phytotoxicity of the isolated compounds on the standard target species Lactuca sativa was evaluated