Antihyaluronidase and Antioxidant Potential of Atriplex sagittata Borkh. in Relation to Phenolic Compounds and Triterpene Saponins.

The genus Atriplex provides species that are used as food and natural remedies. In this work, the levels of soluble phenolic acids (free and conjugated) and flavonoids in extracts from roots, stems, leaves and flowers of the unexplored Atriplex sagittata Borkh were investigated by LC-ESI-MS/MS, together with their antioxidant and antihyaluronidase activity. Phenolic acids were present in all parts of A. sagittata; and were most abundant in the leaves (225.24 µg/g dw.), whereas the highest content of flavonoids were found in the flowers (242.71 µg/g dw.). The most common phenolics were 4-hydroxybenzoic and salicylic acids, kaempferol-3-glucoside-7-rhamnoside, kaempferol-3-rutinoside and the rare narcissoside, which was present in almost all morphotic parts. The stem extract had the highest antioxidant activity and total phenolic content (611.86 mg/100 g dw.), whereas flower extract exerted the most potent antihyaluronidase effect (IC50 = 84.67 µg/mL; control-quercetin: IC50 = 514.28 µg/mL). Phytochemical analysis of the flower extract led to the isolation of two triterpene saponins that were shown to be strong hyaluronidase inhibitors (IC50 = 33.77 and 168.15 µg/mL; control-escin: IC50 = 307.38 µg/mL). This is the first report on the presence of phenolics and saponins in A. sagittata. The results suggest that both groups of metabolites may contribute to the overall activity of this plant species.

Simultaneous Quantification of Ursolic and Oleanolic Acids in Glechoma hederacea and Glechoma hirsuta by UPLC/MS/MS.

The content of ursolic acid and oleanolic acid was determined in different plant parts of two Glechoma species, G. hederacea and G. hirsuta. To achieve optimal extraction conditions of ursolic acid and oleanolic acid from plant material, several methods including maceration, heat reflux, Soxhlet, and ultrasonic extraction, as well as various solvents (methanol, dichloromethane, ethyl acetate), were investigated and compared.For the simultaneous quantification of pentacyclic triterpenes in extracts from Glechoma sp., an UPLC-MS/MS was developed and validated. The method exhibited good linearity, precision, and recovery, and it also was simple, specific, and fast. We developed the method for future application in the quality control of plant materials and botanical extracts containing ursolic acid and oleanolic acid. With regard to the triterpene constituents, both G. hederacea and G. hirsuta can be used equally, and the aboveground parts of both species, but the leaves especially, are abundant sources of ursolic acid (7.1 - 7.5 mg/g dry weight [DW]). Dichloromethane as an extractant provided the best extraction efficiency as well as selectivity to obtain Glechoma extracts rich in triterpenes as compared to methanol and ethyl acetate, regardless of the particular extraction technique. Dry dichloromethane extracts from aerial parts of Glechoma sp. obtained by the heat reflux method resulted in products with a high content of UA (17 - 25% w/w) are considered to be convenient and rich sources of this compound.

Anti-inflammatory activity of saponins from roots of Impatiens parviflora DC.

Two triterpene saponins (IPS-1, IPS-2) for the first time were isolated from the roots of Impatiens parviflora DC. (Balsaminaceae). Their anti-inflammatory activity was evaluated by means of two in vitro models: anti-hyaluronidase and anti-denaturation assays. Both saponins were shown to be potent hyaluronidase inhibitors that affect the enzyme in a dose-dependent manner. The anti-hyaluronidase effect of IPS-2 (IC50 = 286.7 µg/mL) was higher than that of the reference drug: escin (IC50 = 303.93 µg/mL). Both saponins protected bovine serum albumin from heat-induced denaturation in a dose-dependent manner. IPS-1 demonstrated higher anti-denaturation effect (IC50 = 86.7 µg/ml) than IPS-2 (IC50 = 109.76 µg/mL) or the standard drug: acetylsalicylic acid (IC50 = 262.22 µg/mL). In conclusion, potent activity of IPS-1, IPS-2 in both in vitro assays shows that saponins from I. parviflora have anti-inflammatory activity. The obtained results allow to suggest that such compounds may be beneficial in inflammatory conditions, especially associated with excessive degradation of hyaluronic acid.

Enhanced accumulation of harpagide and 8-O-acetyl-harpagide in Melittis melissophyllum L. agitated shoot cultures analyzed by UPLC-MS/MS.

Harpagide and its derivatives have valuable medicinal properties, such as anti-inflammatory, analgesic and potential antirheumatic effects. There is the demand for searching plant species containing these iridoids or developing biotechnological methods to obtain the compounds. The present study investigated the effects of methyl jasmonate (MeJa, 50 µM), ethephon (Eth, 50 µM) and L-phenylalanine (L-Phe, 2.4 g/L of medium), added to previously selected variant of Murashige and Skoog medium (supplemented with plant growth regulators: 6-benzylaminopurine 1.0 mg/L, α-naphthaleneacetic acid 0.5 mg/L, gibberellic acid 0.25 mg/L) on the accumulation of harpagide and 8-O-acetyl-harpagide in Melittis melissophyllum L. agitated shoot cultures. Plant material was harvested 2 and 8 days after the supplementation. Iridoids were quantitatively analyzed by the UPLC-MS/MS method in extracts from the biomass and the culture medium. It was found that all of the variants caused an increase in the accumulation of harpagide. In the biomass harvested after 2 days, the highest harpagide content of 247.3 mg/100 g DW was found for variant F (L-Phe and Eth), and the highest 8-O-acetyl-harpagide content of 138 mg/100 g DW for variant E (L-Phe and MeJa). After 8 days, in some variants, a portion of the metabolites was released into the culture medium. Considering the total amount of the compounds (in the biomass and medium), the highest accumulation of harpagide, amounting to 619 mg/100 g DW, was found in variant F, and the highest amount of 8-O-acetyl-harpagide, of 255.4 mg/100 g DW, was found in variant H (L-Phe, MeJa, Eth) when harvested on the 8th day. These amounts were, respectively, 24.7 and 4.8 times higher than in the control culture, and were, respectively, 15 and 6.7 times higher than in the leaves of the soil-grown plant. The total amount of the two iridoids was highest for variant F (0.78% DW) and variant H (0.68% DW) when harvested on the 8th day. The results indicate that the agitated shoot cultures of M. melissophyllum can be a rich source of harpagide and 8-O-acetyl-harpagide, having a potential practical application. To the best of our knowledge we present for the first time the results of the quantitative UPLC-MS/MS analysis of harpagide and 8-O-acetyl-harpagide in M. melissophyllum shoot cultures and the enhancement of their accumulation by means of medium supplementation with elicitors and precursor.

Design, synthesis, and anticonvulsant activity of some derivatives of xanthone with aminoalkanol moieties.

A series of new xanthone derivatives have been synthesized and evaluated for their anticonvulsant properties in the maximal electroshock, subcutaneous metrazole tests and for neurotoxicity in the rotarod in mice, i.p. and rats, p.o. Compound 9: R,S-2-{2-[(1-hydroxybutan-2-yl]amino)ethoxy}-9H-xanthen-9-one and compound 12: R,S-2-{3-[(1-hydroxybutan-2-yl)amino]propoxy}-9H-xanthen-9-one exerted activity in rats, p.o. 2 and 4 h after administration, respectively. Therefore, metabolic stability of the compounds was evaluated with use of rat microsomes, resulting in half-life t1/2 136 and 108 min, respectively, indicating that either the metabolites are very active or the parent compounds exert ADME properties other than metabolism which influence the late onset of activity.

In vitro anti-denaturation and anti-hyaluronidase activities of extracts and galactolipids from leaves of Impatiens parviflora DC.

The in vitro anti-denaturation and anti-hyaluronidase activities of Impatiens parviflora extracts and isolated galactolipids (MGDG-1, DGDG-1) were investigated. This is the first report on these compounds in I. parviflora. All extracts showed anti-hyaluronidase activity, but only methanolic extract from fresh leaves exhibited significant activity against heat-induced denaturation of BSA in a dose-dependent manner. At 500 µg/mL, the extract and the reference drug showed 79.05% and 99.81% inhibition of protein denaturation, respectively. These results indicate that fresh leaves of I. parviflora may be beneficial in inflammatory conditions, especially those associated with protein denaturation, such as rheumatoid arthritis. The study revealed that only MGDG-1 showed weak activity in anti-denaturation assay but both galactolipids were potent inhibitors of hyaluronidase. MGDG-1 completely inhibited the enzyme activity at the concentration of 127.9 µg/mL. These results indicate the potential of galactolipids in the treatment of diseases associated with the loss of hyaluronic acid.

Photocatalytic degradation of sulfamethoxazole in aqueous solution using a floating TiO2-expanded perlite photocatalyst.

Photocatalytic degradation of an antibiotic, sulfamethoxazole (SMX), in aqueous solution using a novel floating TiO2-expanded perlite photocatalyst (EP-TiO2-773) and radiation from the near UV spectral range was studied. The process is important considering that SMX is known to be a widespread and highly persistent pollutant of water resources. SMX degradation was described using a pseudo-first-order kinetic equation according to the Langmuir-Hinshelwood model. The products of the SMX photocatalytic degradation were identified. The effect of pH on the kinetics and mechanism of SMX photocatalytic degradation was explained.

Synthesis and evaluation of antidepressant-like activity of some 4-substituted 1-(2-methoxyphenyl)piperazine derivatives.

A series of new derivatives of N-(2-methoxyphenyl)piperazine have been synthesized for their affinity toward serotonergic receptors and for their potential antidepressant-like activity. They have been evaluated toward receptors 5-HT1A , 5-HT6 , and 5-HT7 , as well as in vivo in the tail suspension, locomotor activity, and motor co-ordination tests. All the tested compounds proved very good affinities toward 5-HT1A and 5-HT7 receptors. The most promising compound was 1-[(2-chloro-6-methylphenoxy)ethoxyethyl]-4-(2-methoxyphenyl)piperazine hydrochloride, exhibiting affinity toward receptors Ki <1 nm (5-HT1A ) and Ki = 34 nm (5-HT7 ). Antidepressant-like activity (tail suspension test) was observed at 2.5 mg/kg b.w. (mice, i.p.), and the effect was stronger than that observed for imipramine (5 mg/kg b.w.). Sedative activity was observed at ED50 (locomotor test, mice, i.p.) = 17.5 mg/kg b.w. and neurotoxicity was observed at TD50 (rotarod, mice, i.p.) = 53.2 mg/kg b.w.

Minor triterpene saponins from underground parts of Lysimachia thyrsiflora: structure elucidation, LC-ESI-MS/MS quantification, and biological activity.

Two minor triterpene saponins, one previously not reported, were isolated from the underground parts of Lysimachia thyrsiflora L. The structures were determined based on a combination of one- and two-dimensional NMR techniques, and mass spectrometry, as 30-O-beta-D-glucopyranosyl-3beta,16alpha,30-trihydroxy-olean-12-en-28-yl acetate 3-O-beta-D-glucopyranosyl-(1--> 4)-O-[beta-D-glucopyranosyl-(1-->2)-]-O-alpha-L-arabinopyranoside (denoted as thyrsiloside A), and davuricoside L. Quantitative determination of these two compounds, along with other saponins present in L. thyrsiflora underground parts, was performed by a developed UPLC-ESI-MS/MS method, validated according to the ICH guidelines. The relative total content of triterpene saponins amounted to 1.19 microg/mg dw. Both 4 and 5 were present in minor quantities, 0.031 and 0.077 microg/mg dw, respectively. Assays performed on a panel of human cancer cell lines showed antiproliferative activity against prostate carcinoma PC-3 (GI50 after 48 h = 20 and 24 microg/mL, respectively).

Determination of moxifloxacin and its oxidation products with kinetic evaluation under potassium permanganate treatment in acidic solution by ultra-performance liquid chromatography/tandem mass spectrometry.

A simple, sensitive, and reproducible ultra-performance LC method for the determination of moxifloxacin (MOXI) oxidation stability under permanganate treatment in acidic conditions (pH 3.0-6.0) was developed. Besides the MOXI peak [retention time (RT) = 2.58], four additional products (RT = 0.86, 0.91, 1.42, and 1.89) were observed in all conditions tested. The oxidation process followed second-order reaction kinetics and depended upon solution acidity. The highest reaction rate constant was observed at pH 3.0, and this value decreased as the pH was increased to 6.0. The oxidation products were characterized, and their fragmentation pathways, derived from MS/MS data, were proposed. Two of these products were identified as hydroxyl derivatives of MOXI and two others as their oxidation product analogs with molecular ions of 418.4 and 416.4 m/z, respectively.