Isolation of antioxidants from Alchemilla xanthochlora

The aim of this study was to find conditions for isolation of antioxidants from Alchemilla xanthochlora. The solid-liquid extraction of plant material with different solvents (n-hexane, chloroform, ethylacetate, methanol, and water) was used for preparation of crude extracts of antioxidants. For additional purification of crude extract, liquid-liquid extraction in different organic solvents (chloroform, ethylacetate, methanol) and water, and column chromatography with silica gel as stationary phase and mixture of chloroform : methanol as mobile phase were used. The content of active compounds was monitored by photometric method and special TLC analysis using DPPH as basic reagent. TLC-DPPH analysis of herbal extracts showed that antioxidants from Alchemilla xanthochlora were separated into two groups with different polarity. Finally, for the isolation of antioxidants from Alchemilla xanthochlora were successfully used the following procedures: extraction of dried grounded plant material with methanol at 50°C during 1 hour, subsequent liquid-liquid extraction in isopropanol and water and fractionation of isopropanol extract by column chromatography. Fraction with isolated antioxidants showed antioxidant activity 535.2 mg to g of fraction residue.

Polyphenol Extract from Evening Primrose (Oenothera paradoxa) Inhibits Invasion Properties of Human Malignant Pleural Mesothelioma Cells

Extracts from the defatted evening primrose (Oenothera paradoxa Hudziok) seeds are the source of a range of stable polyphenolic compounds, including ellagic acid, gallic acid, and catechin. Our studies evaluate, for the first time, the influence of evening primrose isopropanol extract (EPE) on malignant pleural mesothelioma (MPM) cells. MPM is rarely diagnosed, its high aggressiveness and frequently noted chemoresistance limit its treatment schemes and it is characterized by low prognostic features. Here, we demonstrate that EPE inhibited MPM growth in a dose-dependent manner in cells with increased invasion properties. Moreover, EPE treatment resulted in cell cycle arrest in the G2/M phase and increased apoptosis in invasive MPM cell lines. Additionally, EPE strongly limited invasion and MMP-7 secretion in MPM cancer cells. Our original data provide evidence about the potential anti-invasive effects of EPE in MPM therapy treatment.

THE SIBERIAN LARCH GROUP COMPOSITION AND ACID NEEDLES AT DIFFERENT VEGETATION PERIODS

The research results of the Siberian larch needle extractives are presented in paper. The content of extractive substances from green and yellow needles extracted by organic solvents with different polarities is determined. It is found out that the double increase of the yellow needle extractive substances content soluble in hydrocarbon extractant is observed, whereas it is decreased by 30 per cent when extracted with propan-2-ol. The group composition of needle extracts derived from isopropanol extract by petroleum ether is determined. The neutral substances content in both needle collections is practically the same, although the yellow needles contain less acid “bound” in the form of esters. There is a high content of waxes in the needles, which mainly consist of secondary nonacosanol-10 alcohol. The composition of free and “bound” acids in Siberian larch needles of different vegetation periods is established. The needles are dominated by groups of higher unsaturated C-18 fatty acids. Resin acids are represented by isopimaric acid (isopimaric and sandaracopimaric) and abietic acids: abietic, dehydroabietic and levopiric acids. In yellow needles, the content of dehydroabietic acid is increased quadrupled and the hydroxy acids are identified: 15-hydroxyabietic acid and 15-hydroxyhydroabietic acid. The tri- and tetraenic C-20 acids are identified among higher fatty acids.

Gastroprotective effects of the isopropanol extract of Artemisia princeps and its gastroretentive floating tablets on gastric mucosal injury

In this study, we investigated the gastroprotective effect of an isopropanol extract from the aerial parts of Artemisia princeps (IPAP) and developed a gastroretentive floating tablet of IPAP (IPAP-FR) for maximized local gastroprotective effects. Pre-treatment with IPAP ameliorated the gastric mucosal hemorrhagic lesions in ethanol/HCl- or indomethacin- treated rats. IPAP decreased mucosal hemorrhage of gastric ulcers induced by ethanol or indomethacin plus pyloric ligation in rats. The optimized floating tablet, IPAP-FR, floated on medium surface with more sustained eupatilin release compared to the non-floating control tablet. X-ray photographs in beagle dogs showed that IPAPFR was retained for > 2 h in the stomach. In the ethanol-induced gastric ulcer rat model, the gastric hemorrhagic lesion was improved more substantially with IPAP-FR compared to the non-floating control tablet. Based on these data, our data suggest that IPAP-FR has an improved therapeutic potential for the treatment of gastric ulcer.

Purification and Fractionation of Natural Petroleum Sulfonates by Liquid-Liquid & Soxhlet Extraction

Natural petroleum sulfonates (PS) was separated, purified and divided into several fractions by a new method liquid-liquid & soxhlet extraction. The effects of heating time, volume ratio of extractants (isopropanol aqueous solution-50/50 (by volume) : n-pentane), gradient solvent types, extracting time & times on the separation efficiency are discussed, respectively. And the extracted mineral oil was anlalyzed by IR spectra, the average equivalent weight of each extracted PS fraction was measured by ESI-MS. The experimental results show that the volatiles in natural plant-manufactured PS can evaporate thoroughly in 3.5h at 130°C, and when the volume ratio of stated extractants is 3:1, after several times extraction & anti-extraction, the extracted mineral oil is PS-free, and the purified PS is successfully separated from inorganic salt, meanwhile the purified PS was divided into four fractions, whose average equivalent weight are (toluene extract) 595.03, (dichloromethane extract) 486.00, (isopropanol extract) 474.85 and (alcohol extract) 384.5g/mol, respectively. The proposed method is superior to the traditional liquid-liquid extraction for simplified steps and bifunction with PS purifying & fraction dividing, can separate efficiently industrial PS and other similar mixtures.

Evaluation of Traditional Indian Antidiabetic Medicinal Plants for Human Pancreatic Amylase Inhibitory EffectIn Vitro

Pancreaticα-amylase inhibitors offer an effective strategy to lower the levels of post prandial hyperglycemia via control of starch breakdown. Eleven Ayurvedic Indian medicinal plants with known hypoglycemic properties were subjected to sequential solvent extraction and tested forα-amylase inhibition, in order to assess and evaluate their inhibitory potential on pancreaticα-amylase. Analysis of 91 extracts, showed that 10 exhibited strong Human Pancreatic Amylase (HPA) inhibitory potential. Of these, 6 extracts showed concentration dependent inhibition withIC50values, namely, cold and hot water extracts fromFicus bengalensisbark (4.4 and 125 μgmL-1),Syzygium cuminiseeds (42.1 and 4.1 μgmL-1), isopropanol extracts ofCinnamomum verumleaves (1.0 μgmL-1) andCurcuma longarhizome (0.16 μgmL-1). The other 4 extracts exhibited concentration independent inhibition, namely, methanol extract ofBixa orellanaleaves (49 μgmL-1), isopropanol extract fromMurraya koenigiileaves (127 μgmL-1), acetone extracts fromC. longarhizome (7.4 μgmL-1) andTribulus terrestrisseeds (511 μgmL-1). Thus, the probable mechanism of action of the above fractions is due to their inhibitory action on HPA, thereby reducing the rate of starch hydrolysis leading to lowered glucose levels. Phytochemical analysis revealed the presence of alkaloids, proteins, tannins, cardiac glycosides, flavonoids, saponins and steroids as probable inhibitory compounds.

Lipid Mobilization in Dormant and Nondormant Caryopses of Yellow Foxtail (Setaria lutescens)

The major reserve lipid of yellow foxtail [Setaria lutescens(Weigel) Hubb.] caryopses is triacylglyceride stored in lipid bodies. Lipid bodies are heavily concentrated in the cells of the embryo axis, scutellum, and aleurone layer and are roughly spherical (average diam 0.5μm). Percentage lipid (hot isopropanol extract) on a fresh-weight basis of mature, unimbibed seed and caryopses collected locally was about 6.4 and 11%, respectively. About 95% of the total (0.23 mg/caryopsis) was saponifiable and distributed among fatty acid classes as follows: 16:0, 5.8%; 18:0, 1.5%; 18:1, 16.2%; 18:2, 73.7%; and 18:3, 2.8%. In nondormant caryopses, saponifiable lipid content declined (18:1 and 18:2 only) about 23% during the first 6 days after imbibition in the dark at 25 C. This modest decline was preceded by the appearance of an alkaline lipase at about 24 h after the onset of imbibition. Free fatty acids accumulated during these early stages of germination and accounted for about 22% of the total saponifiable lipid remaining (0.17 mg/caryopsis) 6 days after the onset of imbibition. Lipid content and lipase activity did not change in dormant caryopses treated similarly. The delayed initiation of lipid mobilization in germinating caryopses suggests that triacylglyceride degradation is not a pivotal metabolic pathway controlling dormancy.

Determination of Starch and Soluble Carbohydrates. I. Development of Method for Grains, Stock Feeds, Cereal Foods, Fruits, and Vegetables

Starch is determined in samples containing not > 400 rag available carbohydrate by the following procedure: The fat is extracted with isopropanol, the solution is diluted to 70% alcohol concentration, NaCl and Celite are added, and the soluble sugars are extracted. Starch is hydrolyzed with Rhozyme-S, the solution is clarified with Zn(OH)2 , and the reducing sugar content is determined by reduction of potassium ferricyanide. Maltose and other soluble saccharides are present in the 70% isopropanol extract. The solubility of enzymatically hydrolyzable oligosaccharides, minus maltose, of three commercial soluble starches and three commercial dextrins in 0 - 7 0% isopropanol-1 % NaCl was determined. In 70% isopropanol, the respective solubilities of the hydrolyzable oligosaccharides were as follows: starches—0.4, 3.4, and 0.5%; dextrins—4.9, 3.3, and 6.1%. In 40% isopropanol, the respective solubilities were 12.7, 9.8, and 62.3% of the starches and 92.3, 88.2, and 89.4% of the dextrins. Although the soluble starches and dextrins were water-soluble, they contained up to 9.5% (6.5% average) oligosaccharides that were not hydrolyzable by enzyme but were hydrolyzable by l.OA' HCl. Use of 40% isopropanol is recommended for extraction of oligosaccharides of low DP (dextrins, maltose, etc.) and determination of the insoluble oligosaccharides of relatively high DP. Retrogradation of starch and its prevention during the analysis is discussed. Starch and soluble sugars were determined in 21 samples of grains and breakfast cereals and in 12 samples of roughages.