A kind of kudzu root polysaccharide and its preparation method and application
A technology of pueraria polysaccharides and polysaccharides is applied in the preparation of lipid-lowering drugs, the preparation of pueraria homogeneous polysaccharides, and the field of polysaccharides, and can solve the problems of unclear medicinal substances and the like.
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Embodiment 1
[0019] Example 1: Preparation of kudzu root polysaccharide QL
[0020] Pueraria 7.0Kg is pulverized, extracted 3 times with 90°C water at a solid-to-liquid ratio of 1:20, filtered, combined with extracts, and concentrated to a density of 1.1-1.2; then add 2 times the volume of absolute ethanol and let it stand for about 3 hours , filtered to obtain a precipitate, redissolve the precipitate in water, add 0.1% activated carbon to remove the pigment, centrifuge, concentrate the supernatant to 0.5-1L, freeze-dry to obtain fluffy kudzu root polysaccharide 100-244g; weigh Add 200g of crude polysaccharide with distilled water and magnetically stir to dissolve, centrifuge, add the supernatant to a macroporous resin column HP-20 for separation, elute with pure water, 10% ethanol, and 20% ethanol solution, collect each fraction, and analyze according to sulfuric acid- The same fractions were combined by phenol color development and 490 nm detection results, then concentrated, dialyzed a...
Embodiment 2
[0023] Example 2: Structural characterization of kudzu root polysaccharide (QL)
[0024] (1) Determination of molecular weight
[0025] Using TSK-GEL GMPWXL gel column (300×7.6 mm), the mobile phase is 0.01 mol / L NaNO3, the flow rate is 0.8 mL / min, the column temperature is 25°C, and a series of molecular weight dextran is used as standard music. The measured kudzu root Polysaccharide QL has a molecular weight of 10-60 KDa.
[0026] (2) Determination of total sugar, uronic acid and protein content
[0027] The sulfuric acid-phenol method determined the total sugar content of QL to be 98.7%;
[0028] The uronic acid content of QL measured by m-hydroxybiphenyl method is 0.0%;
[0029] The protein content of QL determined by Coomassie Brilliant Blue method was 0.64%;
[0030] (3) Sugar composition analysis
[0031] QL was hydrolyzed by 2.0 mol / L TFA at a constant temperature of 100°C for 6 hours, and the product obtained by complete hydrolysis was reduced by adding an approp...
Embodiment 3
[0036] Embodiment 3: In vivo lipid-lowering experimental method
[0037] Fifty-six male Wistar rats were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. (8 weeks, body weight 200±20 g). All animals were fed with normal diet for 1 week. Subsequently, 8 rats were used as the blank control group (abbreviated as the blank group (1) for short), fed with normal diet, and the remaining 48 rats were fed with high-fat diet (HF, 88.5% of common rat diet with 1.2% cholesterol, 0.3% cholic acid sodium, 10% lard) for 2 months to establish a hyperlipidemia model.
[0038]Rat plasma levels of TG, TC, LDL-C and HDL-C were sampled every half month. After 2 months, all rats were anesthetized with isoflurane and blood samples were collected from orbital veins using capillary tubes. Centrifuge the blood sample (4°C; 4,000 r×min-1; 15min) to obtain plasma samples, use the kit (purchased from Nanjing Jian Biotechnology Co., Ltd., China) to make a standard curve, and ...
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