Oct 3, 2017 - Polysaccharides in blackberry seeds were extracted, purified and identified. Extraction and purification of polysaccharide.
The blackberry seed was typically removed as a byproduct and waste from blackberry fruits for juices. Developing value-added utilization of berry seeds will significantly expand the market for berry products as well as improve benefit to berry producers. However, the research on blackberry seed is limited.
The objective of this paper was to research antithrombotic mechanism of polysaccharides in blackberry seeds. Polysaccharides in blackberry seeds were extracted, purified and identified by high-performance gel permeation chromatography (HPSEC), gas chromatography (GC), fourier transform infrared (FT-IR) spectrometer and nuclear magnetic resonance spectra (NMR).
Anticoagulant activities were evaluated in vivo by measuring activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin time (PT), fibrinogen (FIB) and plasma recalcification time (RRT). Four polysaccharides named BSP-1a, BSP-1b, BSP-2 and BSP-3 were isolated from Blackberry ( Rubus spp.) seeds. The results indicated that BSP-1b, BSP-2 and BSP-3 exhibited the anticoagulant activity.
Therefore, the anti-thrombosis effects of BSP-1b, BSP-2 and BSP-3 were investigated in vivo by 6-Keto-PGF 1α, thromboxane B 2 (TXB 2), endothelial nitric oxide synthase (eNOS), endothelin-1 (ET-1), whole blood viscosity (WBV), plasma viscosity (PV), hematocrit (Hct), erythrocyte sedimentation rate (ESR), APTT, TT, PT and FIB. The results suggested that BSP-1b, BSP-2 and BSP-3 had the inhibition effect on thrombus formation, and the antithrombotic effects were associated with the regulation of vascular endothelium active substance, activating blood flow and anticoagulation effect.
Introduction Blackberry ( Rubus spp.) is a widely distributed shrub with high commercial value due to its sensory and chemical characteristics,such as sugars, malic acid, amino acids, VC, VE, anthocyanins, superoxide dismutase (SOD) and flavonoids []. Therefore, it is usually used as raw material for the production of jams, candy, wine and juice beverages, and so on. The seeds of the blackberry fruits were typically removed during juice processing. In our previous research, 13 compounds were isolated and identified from blackberry seeds, pharmacology research indicated that blackberry seeds had antioxidant [], cholesterol-lowering [], liver-protection effects [], and could rectify the derangement in lipide metabolism [].
Font collection mac torrent. However, there is no research on polysaccharides from blackberry seeds in literature. Microsoft encarta 2016 free download full version. Plant polysaccharides display a wide variety of biological activity, including immunostimulatory [], anti-cancer [], anti-diabetic [] and antithrombotic activities []. As already shown for algae polysaccharides, so do plant polysaccharides have anticoagulant activity [–], and can be used as a prevention and treatment of cardiovascular disease []. The aim of this study was to isolate, characterize and evaluate the anticoagulant and antithrombotic effects of the polysaccharides of blackberry seeds.
The results may provide a reference for the further research and development of Blackberry ( Rubus spp.). Extraction and purification of polysaccharide The dried Blackberry seeds (200 g) were ground into powder and defatted three times by petroleum ether at room temperature. The organic solvent was volatilized to obtain a dry powder. Then the powder was extracted three times with 70% ethanol and filtered. Finally, the residue was lyophilized.
Subsequently, the dried powder was extracted with 20 volumes of distilled water at 80° every 3 h for three times. The aqueous extract was filtered and the supernatant was treated with 95% ethanol (final concentration 70%) at 4° overnight, and centrifuged at 10,000 rpm for 10 min. The precipitation was added with Sevage reagent (chloroform/1-butanol, 1:4 v/v) for deproteinization deproteinisation []. The crude polysaccharide was obtained through precipitation with 95% ethanol (final concentration 70%) and centrifuged. Then the precipitate was redissolved in water and dialyzed for 2 days to remove small molecular weight impurity (molecular weight cut off 3500 Da). Finally, the aqueous extract was lyophilized in vacuum to give the crude polysaccharide (9.7 g). The 300 mg crude polysaccharides were dissolved in 10 mL distilled water, and then the water was filtered through 0.45 μm microporous membrane.
Then the filtered water was fractioned by DEAE-52 column (2.5 × 60 cm). The column was eluted with distilled water at 0.8 mLmin −1 followed by 0.1 M NaCl and 0.2 M NaCl, respectively. The fractions were collected using an automated step-by-step fraction collector and guided for total carbohydrate using the phenol-sulfuric acid method []. Three main fractions were collected, dialyzed, lyophilized and named as BP-1(74 mg), BP-2 (104 mg)and BP-3(95 mg).