Anemarrhena polysaccharide and its preparation method, identification method and application
A technology of Anemarrhena and polysaccharide, applied in the field of Anemarrhena polysaccharide and its preparation, can solve the problems such as insufficient research on the structure of Anemarrhena polysaccharide, and achieve the effects of enhanced neuroprotective activity, clear structure and significant effect
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Embodiment 1
[0048] Embodiment 1 The preparation method of Anemarrhena polysaccharide
[0049] The preparation method of described Anemarrhena polysaccharide comprises the following steps:
[0050] S1 Cutting: Cut 5 kg of dry root of Zhimu to 1~3 cm with scissors, wash it quickly with cold water, dry it, and get the mother section;
[0051] S2 water extraction: add water 10 times its weight to the mother section obtained in step S1, heat to 80°C for extraction, extract for 3 hours, collect the extract, dry the dregs to obtain the extract and dregs;
[0052] S3 graded alcohol precipitation:
[0053] Concentrate the extract obtained in step S2 under reduced pressure at 60°C, add ethanol until the volume concentration of ethanol is 50%, let it stand at room temperature for 24 hours, centrifuge, collect the precipitate and supernatant, and obtain crude polysaccharide AA1 and supernatant 1;
[0054] After the supernatant 1 was concentrated under reduced pressure at 60°C, ethanol was added until...
Embodiment 2
[0057] Example 2 Anemarrhena polysaccharide AA2-1
[0058] The Anemarrhena polysaccharide AA2-1 is obtained by secondary purification of the polysaccharide AA1 obtained in Example 1, and the specific method is as follows:
[0059] 1) Ion-exchange column chromatography: take 200 mg of the parent polysaccharide AA2 obtained in Example 1, dissolve it in 5 mL of deionized water, load it on a DEAE-Sepharose FF column, and one appears under the eluent conditions of different salt concentrations peak, where the elution peak is the 0.1M NaCl elution part (the elution curve is tracked by the phenol-sulfuric acid method during the elution process, and the sugar part is collected separately according to the elution curve), and the obtained eluate is concentrated and freeze-dried , to obtain a polysaccharide;
[0060] 2) Molecular sieve gel chromatography: dissolve the above-mentioned freeze-dried polysaccharide sample in water, centrifuge, take the supernatant, put it on a Sephadex G-75...
Embodiment 3
[0061] Example 3 Structural analysis of Anemarrhena polysaccharide AA2-1
[0062] (1) Test material: Anemarrhena polysaccharide AA2-1.
[0063] (2) Test method:
[0064] 1. Monosaccharide composition analysis (PMP pre-column derivatization method)
[0065] Sample handling:
[0066] Accurately weigh 4.0 mg of the test material into a stoppered test tube, add 2.0 mL of 2 M trifluoroacetic acid (TFA), and place it in an oil bath for hydrolysis at 120 °C for 6 h. Cool to room temperature, repeatedly add methanol and spin dry, remove TFA, dissolve to 1 mL with deionized water, centrifuge, absorb 100 mu L sample solution, add 100 mu L 0.3 M NaOH solution, then add 100 mu L 0.5 MPMP methanol solution, mix well, react on 70°C water bath for 30 min, cool, add 105 mu Neutralize with L 0.3 M HCL solution, add deionized water to 1 mL, then add an equal volume of chloroform solution, shake vigorously, centrifuge, remove the chloroform phase, repeat this extraction twice, take the...
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