Achyranthes bidentata polysaccharide as well as preparation method and application thereof
A polysaccharide of Achyranthes and Achyranthes giganta are applied in the field of polysaccharide of Achyranthes giganta, which can solve the problems of low polysaccharide content and complex composition of total polysaccharides, and achieve the effects of simple preparation method, convenient process expansion and mild reaction conditions.
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Embodiment 1
[0030] Preparation method of refined Achyranthes bidentata polysaccharide (AB)
[0031] After pulverizing the Achyranthes bidentata medicinal material, weigh 500g of medicinal material powder and add 5 times of absolute ethanol, condense and reflux extraction at 80-90°C for 3 times, each time for 2 hours, remove small molecular impurities, filter, collect medicinal residues, and dry at 50°C. Add 10 times the volume of distilled water, condense and reflux at 70-80°C for 3 times, each time for 2 hours, filter the combined filtrate, concentrate under reduced pressure at 45-55°C to about 1.0L, add 4 times the volume of absolute ethanol to precipitate, and stand overnight at 4°C , remove the upper clarified solution to collect the precipitate, wash with absolute ethanol, acetone, and absolute ether successively, add an appropriate amount of distilled water to dissolve the polysaccharide precipitate, then use the Savage method to repeatedly deproteinize, collect the Achyranthes biden...
Embodiment 2
[0033] Preparation method of Achyranthes bidentata homogeneous polysaccharide AB-1-1
[0034] Weigh 3.0 g of Achyranthes bidentata polysaccharide dry powder and add appropriate amount of distilled water to prepare a saturated solution, conduct primary separation through DEAE-FF column (60×2.6cm), and carry out gradient washing with distilled water, 0.1, 0.2, 0.3, 0.4, 0.5M NaCl solution successively Elution, elution flow rate 2.0-3.0mL / min, automatic collection at 40-50mL / T, color development by sulfuric acid-anthrone method, ultraviolet detection in the range of 620-630nm, the absorbance value is the ordinate, and the number of collection tubes is the abscissa Draw the elution curve, collect the samples according to the elution curve, and freeze-dry; the components eluted with NaCl solution need to be dialyzed through the dialysis bag for 48 hours and then freeze-dried.
[0035] Take 400 mg of the above freeze-dried Achyranthes bidentata polysaccharide powder, dissolve in 2.0...
Embodiment 3
[0037] Determination of physicochemical properties of Achyranthes bidentata homogeneous polysaccharide AB-1-1
[0038] 1. Determination of purity and molecular weight
[0039] High performance gel permeation chromatography: Agilent 1290HPLC system equipped with G4260BELSD detector, AgilentPL aquagel-OH MIXED-H, Agilent PL aquagel-OH MIXED-M (300×7.5mm, 8um) series column, 25℃, 0.6mL / min ultra Pure water mobile phase detection, sample concentration 1.0mg / mL, injection 10uL, detection time 40min, such as figure 2 As shown, the results show that AB-1-1 is a high-purity homogeneous polysaccharide;
[0040]The SEC-MALS system was used to measure the weight average molecular weight, number average molecular weight and intrinsic viscosity of AB-1-1, 1200HPLC combined with MALS, ViscoStar, dRI detector, Shodex OHpak-806M HQ chromatographic column, physiological saline as mobile phase, sample concentration 4.5 mg / mL, inject 100uL. Such as image 3 and Figure 4 As shown, the resu...
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