Sansevieria trifasciata polysaccharides as well as preparation method and application thereof
A technology of tiger piranha and blue polysaccharides, applied in the field of tiger piranha polysaccharides and its preparation, can solve the problems that there is no major breakthrough in the structure and structure-activity relationship of polysaccharides
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Embodiment 1
[0037] 1) Take tiger Pilan dry powder, extract with petroleum ether, reflux at 90°C until the extract is almost colorless, and then reflux with 80% ethanol at 60°C for 2h.
[0038] 2) Use DK-98-1 constant temperature water bath (Tianjin Taisite Instrument Co., Ltd.) to extract, under the conditions of extraction temperature of 90°C, extraction time of 160min, and solid-liquid ratio of 1:30, hot water extraction is used.
[0039] 3) The extract was concentrated under reduced pressure by a RE-52C rotary evaporator (Shanghai Qingpu Instrument Factory), and 3 times the volume of 95% ethanol was added to precipitate overnight at 4°C.
[0040] 4) The precipitate was centrifuged in a DL-5-B centrifuge (Shanghai Anting Scientific Instrument Factory), washed with absolute ethanol, acetone, and anhydrous ether in sequence, and finally passed through a FD-1B-50 vacuum freeze dryer (Beijing Boyikang) Experimental Instrument Co., Ltd.) freeze-dried to obtain the crude polysaccharide of Tiger...
Embodiment 2
[0041] Example 2 The crude polysaccharide obtained by hot water extraction is deproteinized by enzymatic method combined with Sevage method
[0042] 1) Adjust the pH of the polysaccharide solution to be deproteinized to 8.0, add trypsin in an amount of 2%, act at 37°C for 2 hours, stir at intervals, and then rise to 80°C to inactivate the enzyme.
[0043] 2) Centrifuge the treated extract at 4000r / min for 10min, take the supernatant and concentrate, deproteinize by Sevag method, repeat 3 times.
[0044] 3) Use the dialysis method to dialyze the deproteinized tiger Piran polysaccharide concentrate with distilled water for 48 hours at 4°C, change it several times in the middle, concentrate after dialysis, precipitate with 3 times 95% ethanol, centrifuge and vacuum freeze-dry to obtain preliminary Purification of Tiger Piran polysaccharide TPS.
Embodiment 3
[0045] Example 3 The deproteinized polysaccharide is purified by DEAE-52 column chromatography:
[0046] 1) Select a chromatographic column with a specification of 3.2cm×40cm, use 3-5 times the column volume of deionized water to run the column under constant pressure, and pass through the chromatographic column at a flow rate of 1ml / min.
[0047] 2) Take the deproteinized crude polysaccharide and dissolve it in a small amount of distilled water, use 0.5g sample volume, and the sample volume is 10ml.
[0048] 3) Elute with deionized water and 0.05, 0.1, 0.2, 0.3mol / L NaCl solution in sequence, control the flow rate at 0.75ml / min, and collect the eluate with SBS-100 automatic partial collector (Shanghai Huxi Instrument Factory), Collect approximately 10ml per tube.
[0049] 4) Combine and collect the elution peaks of I, II, III, IV, and V respectively, concentrate, and dialyze to AgNO 3 Vacuum freeze-dry until no white precipitate is detected. The five elution peaks are re...
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