Method for rapidly preparing linseed polysaccharide with antiviral and immunomodulatory activities
An immunoregulatory activity, flaxseed technology, applied in the fields of biomedicine and functional food, can solve the problems of low efficiency of flaxseed polysaccharide, etc., and achieve the effects of high protein removal efficiency, mild conditions and simple method.
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Embodiment 1
[0031] Embodiment 1: Preparation of flaxseed polysaccharide FHP-2
[0032] A method for rapidly preparing flaxseed polysaccharides with antiviral and immunoregulatory activities, comprising the following steps:
[0033] 1) Separation of linseed husks and kernels: After cleaning and removing impurities, linseeds are fully crushed to obtain linseed powder, which is washed twice with ethanol and then sieved by wet method, and the intercepted part of the sieve is taken and fully dried to obtain linseed husks. The conditions for ethanol washing are: washing temperature 50°C, washing time 30 minutes, solid-liquid ratio 1:4, continuous stirring during the alcohol washing process; the specific operation of wet sieving is: fully stir the alcohol washing system mixture to make the solid material uniform Suspend in the ethanol system, then quickly pass the mixed material through a 425μm sieve, so that the ethanol extract and fine linseed kernel particles pass through the sieve holes, and...
Embodiment 2
[0037] Example 2: Characterization of flaxseed polysaccharide FHP-2
[0038] (1) Determination of relative molecular mass
[0039] FHP-2 was dissolved in ultrapure water to prepare a 2.5 mg / mL sample solution, and its purity and molecular weight were determined by GPC. The specific test conditions and methods are as follows: using TSK G-5000PW XL and TSK G-3000PW XL The chromatographic columns are connected in series, the 2414 type differential refractive index detector, the column temperature is 35°C, and the mobile phase is 0.02mol / L KH 2 PO 4 , the flow rate is 0.6mL / min. The results showed that the components of FHP-2 were uniform and the relative molecular weight was 1182kDa.
[0040] (2) Monosaccharide composition analysis
[0041] Hydrolysis of polysaccharide samples: Take 10mg of FHP-2 samples, add 4.0mL of 2mol / L trifluoroacetic acid to hydrolyze at 110°C for 8 hours, spin the hydrolyzate to dry, wash with methanol for 3 times, and wash with N while hot. 2 blow...
Embodiment 3
[0057] Embodiment 3: the anti-hepatitis B virus activity of FHP-2
[0058] (1) MTT experiment
[0059]Take the activated HepG2.2.15 cells in the logarithmic growth phase at 1×10 5 seed / mL concentration in 96-well plate, placed in 5% CO 2 , cultured at 37°C, and after 24 hours, 150 μL of the drug group added with FHP-2 was replaced, and the group without drug was used as a blank control, and the culture was continued. After 24 hours, 20 μL of MTT solution was added, the culture was terminated after continuing to cultivate for 4 hours, and the culture solution in the well was sucked off. Add 150 μL of dimethyl sulfoxide to each well, shake on a shaker at low speed for 10 minutes, and fully dissolve the crystals. Measure the absorbance value at a wavelength of 450nm with an enzyme-linked immunosorbent detector, and calculate the cell survival rate (SR) according to the following formula:
[0060] Survival rate SR (%) = A 1 / A 0 ×100%, where A 1 is the absorbance value of t...
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