Hepatitis B virus resistant active composition of polyprenols from ginkgo biloba leaves and preparation method thereof
A technology of ginkgo biloba polyprenol and polyprenol, which is applied in the direction of antiviral agents, drug combinations, organic active ingredients, etc., can solve the problems of large loss, high cost, and low yield of polyprenol, and achieve Good inhibitory effect and yield improvement effect
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Embodiment 1
[0035] The HPLC method of embodiment 1 polyprenol
[0036] This implementation is analyzed by HPLC, the chromatographic column is Kromasil C18 ODS-1 (5 μm, 150×4.6mm), the column temperature is room temperature, the ultraviolet detection wavelength is 210nm, the mobile phase is isopropanol:methanol=38:37, and the flow rate is 1.0mL / min. C95polyprenol was used as the standard control substance with a purity of 98% (provided by Larodan Fine Chemicals Company, Sweden).
[0037] (1) Preparation of standard reference solution
[0038] Accurately weigh 4.0 mg of polyprenol standard reference substance (C95 polyprenol) with an analytical balance, dissolve it in n-hexane, and set the volume to 10 mL. The concentration of the prepared standard solution is 0.40 mg / mL.
[0039] (2) Drawing of standard reference substance curve
[0040] Draw 1 μL, 2 μL, 3 μL, 4 μL and 5 μL of polyprenol standard reference solution (0.4 mg / mL) with a micro-syringe, inject samples according to the above...
Embodiment 290
[0052] Embodiment 290% ginkgo biloba polyprenol preparation method
[0053] Ginkgo biloba fat-soluble matter with a polyprenyl alcohol content of less than 40% is used as raw material, dissolved in petroleum ether at a solid-to-liquid ratio (g / mL) of 1: (1-30), and activated carbon and attapulgite are used as a mixture The decolorizing agent has a mass ratio of 1: (1-20), the mass ratio of the decolorizing agent to the crude polyprenol is (10-150): 100, and is stirred and decolorized at 50-120° C. for 10-80 minutes. Filtrate for the second time, combine the decolorized supernatant, and recover the solvent under vacuum at room temperature to prepare decolorized polyprenol. Adsorb the decolorized polyprenyl alcohol and silica gel at a mass ratio of 1: (15-50), the eluent is petroleum ether: ethyl acetate = 100: (1-30) mixed solvent, the silica gel column length is 20-300cm, the column The diameter is 2-30cm, the column pressure is 0.3-3MPa, the detection wavelength is 199-220nm...
Embodiment 3
[0054] The medium-pressure silica gel column purification method of embodiment 3 polypentenol
[0055] Take 20g of vegetable fat-soluble unsaponifiable matter of 25% polyprenol, add 250mL of n-hexane: ethyl acetate: acetone = 8:3:89 mixed solvent, freeze at -30°C for 3h, centrifuge and filter at -20°C, Once, the supernatants were combined, and the solvent was recovered in vacuo at room temperature to prepare 16 g of polyprenol ointment. Get 10g of polypentenol ointment with a purity of 31.8%, dissolve it in 100mL of petroleum ether, use activated carbon and attapulgite as a mixed decolorizing agent, the amount of decolorizing agent is 12g, the ratio of activated carbon to attapulgite is 1:5, and the decolorization temperature is 70 °C, stirred for 20 minutes to decolorize. After decolorization, the purity increased to 49.6%. Take 400 mesh of silica gel, dry-pack the column (column length 30cm, column diameter 2.5cm), pump air at -0.05 to -0.08MPa for 20-30min, add petroleum ...
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