A method for preparing quercetin-3-o-2", 6"-dirhamnosyl glucoside from ginkgo biloba extract

Abstract

The present invention discloses a method for preparing quercetin-3-O-2",6"-dirhamnosylglucoside from a ginkgo leaf extract. The method comprises: dissolving a ginkgo leaf extract into an ethanol-water solution to prepare a ginkgo leaf extract solution; carrying out one-dimensional liquid chromatography separation on the extract solution to obtain a one-dimensional crude product; dissolving the one-dimensional crude product in a methanol-water solution or ethanol-water solution to obtain a crude product solution; and carrying out two-dimensional liquid chromatography preparation on the crude product solution to obtain the quercetin-3-O-2",6"-dirhamnosylglucoside. According to the present invention, the clear medicinal compound quercetin-3-O-2",6"-dirhamnosylglucoside is prepared from the ginkgo leaf extract, the preparation method is simple, the separation operation is convenient, and the specific compound is separated by using the two-dimensional liquid chromatography method, such that the index component is increased from the ginkgo leaf extract quality control research, the detection object of the active substance is enriched, and the quality standards of the ginkgo leaf extract and the drug can be easily improved.

Description

technical field

[0001] The present invention relates to a method for extracting and preparing an active ingredient in Ginkgo biloba extract, in particular to a method for preparing quercetin-3-O-2", 6"-dirhamnosyl glucoside from Ginkgo biloba extract , belongs to the technical field of compound preparation. Background technique

[0002] Ginkgo biloba is the dried leaf of Ginkgo biloba plant, Ginkgo biloba extract is the enriched product of active substances extracted from Ginkgo biloba through modern extraction technology, which can be used for Alzheimer's disease, depression, diabetes, neurological diseases, impotence , memory impairment, peripheral vascular disease, intermittent claudication, brain turning tinnitus and other diseases. Its main active ingredients are flavonoids and terpenes. Flavonoids include monoflavones, flavonol glycosides, acetylated flavonol glycosides, biflavones, flavan-3-ols, and proanthocyanidins. Terpenoid ginkgolides include ginkgolides A, B,...

Examples

Embodiment 1

[0030]Weigh 10 g of Ginkgo biloba extract, dissolve it in 50 mL of 40% ethanol-water solution to obtain a Ginkgo biloba extract solution with a concentration of 200 mg / mL, pass it through a 0.45 μm microporous membrane, and perform one-dimensional liquid chromatography separation. The one-dimensional liquid chromatography adopts the hydrophilic chromatographic filler 50×250mm with silica gel as the substrate, 10 μm (Huapu New Technology Co., Ltd.), the mobile phase adopts acetonitrile (containing 0.1% formic acid) as the organic phase, water (containing 0.1% Formic acid) is the water phase, and the gradient elution method: the concentration of the organic phase is reduced from 95% to 90% in step gradient for 0-15min. The DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is room temperature, the injection volume is 500μL / needle, the flow rate of the mobile phase is 90mL / min, the fractions collected for 28-30 minutes are co...

Embodiment 2

[0040] Weigh 1 g of Ginkgo biloba extract, dissolve it in 100 mL of ethanol-water solution with a volume concentration of 80%, and obtain a Ginkgo biloba extract solution with a concentration of 10 mg / mL, pass it through a 0.45 μm microporous membrane, and perform one-dimensional liquid chromatography. One-dimensional liquid chromatography adopts silica gel-based hydrophilic chromatographic packing (50×250mm, 10μm, Huapu Innovation Technology Co., Ltd.), the mobile phase uses ethanol as the organic phase, water as the aqueous phase, and the concentration of the organic phase is 90 % isocratic, using a DAD ultraviolet detector at 360nm to select the absorption wavelength, the preparation temperature is 40°C, the injection volume is 200 μL / needle, the flow rate of the mobile phase is 60 mL / min, the fractions collected for 28-32 minutes are concentrated by rotary evaporation to Dry, for one-dimensional preparation of crude quercetin-3-O-2",6"-dirhamnosyl glucoside. Dissolve the c...

Embodiment 3

[0043] Weigh 100g of Ginkgo biloba extract, dissolve it in 200mL of 50% ethanol-water solution to obtain a Ginkgo biloba extract solution with a concentration of 500mg / mL, pass it through a 0.45μm microporous membrane, and prepare it by one-dimensional liquid chromatography. One-dimensional liquid chromatography adopts silica gel-based hydrophilic chromatographic packing (50×250 mm, 10 μ, Huapu New Innovation Technology Co., Ltd.), mobile phase uses ethanol (containing 0.1% formic acid) as the organic phase, water (containing 0.1% formic acid) as the aqueous phase, eluted with 95% organic equivalence. The DAD ultraviolet detector was used to select the absorption wavelength at 360nm, the preparation temperature was 30°C, the injection volume was 3000 μL / needle, the flow rate of the mobile phase was 120mL / min, the fractions collected for 28-32 minutes were concentrated to dryness by rotary evaporation, and obtained as a Preparation of crude quercetin-3-O-2”, 6”-dirhamnosyl gluc...

technical field

[0001] The present invention relates to a method for extracting and preparing an active ingredient in Ginkgo biloba extract, in particular to a method for preparing quercetin-3-O-2", 6"-dirhamnosyl glucoside from Ginkgo biloba extract , belongs to the technical field of compound preparation. Background technique

[0002] Ginkgo biloba is the dried leaf of Ginkgo biloba plant, Ginkgo biloba extract is the enriched product of active substances extracted from Ginkgo biloba through modern extraction technology, which can be used for Alzheimer's disease, depression, diabetes, neurological diseases, impotence , memory impairment, peripheral vascular disease, intermittent claudication, brain turning tinnitus and other diseases. Its main active ingredients are flavonoids and terpenes. Flavonoids include monoflavones, flavonol glycosides, acetylated flavonol glycosides, biflavones, flavan-3-ols, and proanthocyanidins. Terpenoid ginkgolides include ginkgolides A, B,...

Examples

Embodiment 1

[0030]Weigh 10 g of Ginkgo biloba extract, dissolve it in 50 mL of 40% ethanol-water solution to obtain a Ginkgo biloba extract solution with a concentration of 200 mg / mL, pass it through a 0.45 μm microporous membrane, and perform one-dimensional liquid chromatography separation. The one-dimensional liquid chromatography adopts the hydrophilic chromatographic filler 50×250mm with silica gel as the substrate, 10 μm (Huapu New Technology Co., Ltd.), the mobile phase adopts acetonitrile (containing 0.1% formic acid) as the organic phase, water (containing 0.1% Formic acid) is the water phase, and the gradient elution method: the concentration of the organic phase is reduced from 95% to 90% in step gradient for 0-15min. The DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is room temperature, the injection volume is 500μL / needle, the flow rate of the mobile phase is 90mL / min, the fractions collected for 28-30 minutes are co...

Embodiment 2

[0040] Weigh 1 g of Ginkgo biloba extract, dissolve it in 100 mL of ethanol-water solution with a volume concentration of 80%, and obtain a Ginkgo biloba extract solution with a concentration of 10 mg / mL, pass it through a 0.45 μm microporous membrane, and perform one-dimensional liquid chromatography. One-dimensional liquid chromatography adopts silica gel-based hydrophilic chromatographic packing (50×250mm, 10μm, Huapu Innovation Technology Co., Ltd.), the mobile phase uses ethanol as the organic phase, water as the aqueous phase, and the concentration of the organic phase is 90 % isocratic, using a DAD ultraviolet detector at 360nm to select the absorption wavelength, the preparation temperature is 40°C, the injection volume is 200 μL / needle, the flow rate of the mobile phase is 60 mL / min, the fractions collected for 28-32 minutes are concentrated by rotary evaporation to Dry, for one-dimensional preparation of crude quercetin-3-O-2",6"-dirhamnosyl glucoside. Dissolve the c...

Embodiment 3

[0043] Weigh 100g of Ginkgo biloba extract, dissolve it in 200mL of 50% ethanol-water solution to obtain a Ginkgo biloba extract solution with a concentration of 500mg / mL, pass it through a 0.45μm microporous membrane, and prepare it by one-dimensional liquid chromatography. One-dimensional liquid chromatography adopts silica gel-based hydrophilic chromatographic packing (50×250 mm, 10 μ, Huapu New Innovation Technology Co., Ltd.), mobile phase uses ethanol (containing 0.1% formic acid) as the organic phase, water (containing 0.1% formic acid) as the aqueous phase, eluted with 95% organic equivalence. The DAD ultraviolet detector was used to select the absorption wavelength at 360nm, the preparation temperature was 30°C, the injection volume was 3000 μL / needle, the flow rate of the mobile phase was 120mL / min, the fractions collected for 28-32 minutes were concentrated to dryness by rotary evaporation, and obtained as a Preparation of crude quercetin-3-O-2”, 6”-dirhamnosyl gluc...

Images