A method for preparing kaempferol-3-o-p-coumaroyl glucorhamnoside from ginkgo biloba extract

Abstract

The invention discloses a method for preparing kaempferol-3-O-p-coumaric acyl glucosyl rhamnoside from a folium ginkgo extract, wherein the method includes the steps: dissolving the folium ginkgo extract in an ethanol-aqueous solution, to prepare a folium ginkgo extract solution; carrying out one-dimensional liquid chromatography separation of the extract solution, to obtain a one-dimensional crude product; dissolving the one-dimensional crude product in a methanol-aqueous solution or an ethanol-aqueous solution, to obtain a crude product solution; and carrying out two-dimensional liquid chromatography preparation of the crude product solution, to obtain the kaempferol-3-O-p-coumaric acyl glucosyl rhamnoside. The definite medicinal compound kaempferol-3-O-p-coumaric acyl glucosyl rhamnoside is prepared from the folium ginkgo extract; the preparation method is simple, the separation operation is convenient, the particular compound is separated by the two-dimensional liquid chromatography, and thus the index component is increased in research of quality control of the folium ginkgo extract, a detection object of an active substance is enriched, and improvement of quality standards of the folium ginkgo extract and medicines thereof is facilitated.

Description

technical field

[0001] The invention relates to a method for extracting and preparing an active ingredient in ginkgo leaf extract, in particular to a method for preparing kaempferol-3-O-p-coumaroyl glucorhamnoside from ginkgo leaf extract, which 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, C, J,...

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. One-dimensional liquid chromatography adopts silica gel as the matrix of hydrophilic chromatographic filler 50 × 250mm, 10 μm (Huapu New Innovation Technology Co., Ltd.), and the mobile phase adopts methanol containing 0.1% volume concentration of formic acid as the organic phase, containing 0.1% volume The water with formic acid concentration is the water phase, and the gradient elution method: the concentration of the organic phase decreases from 95% to 90% in step gradient in 0-15min. Use DAD ultraviolet detector 360nm to select the absorption wavelength, the preparation temperature is room temperature, the injection volume is 500μL / needle, the mobile phase flow rate is 90mL / min, the fractions coll...

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 200μL / needle, the flow rate of the mobile phase is 60mL / min, the fractions collected for 15-20 minutes are concentrated to dryness by rotary evaporation , for the one-dimensional preparation of crude kaempferol-3-O-p-coumaroyl glucorhamnoside. Dissol...

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 a hydrophilic chromatographic packing (50×250mm, 10 μ, Huapu Innovation Technology Co., Ltd.) based on silica gel, and the mobile phase adopts ethanol (containing 0.1% formic acid) as the organic phase, water (containing 0.1% % formic acid) is the aqueous phase, and is eluted by 95% organic equivalence. The DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is 30°C, the injection volume is 3000μL / needle, the flow rate of the mobile phase is 120mL / min, the fractions collected for 15-20 minutes are concentrated to dryness by rotary evaporation, and the one-dimensional Preparation of crude kaempferol-3-O-p-cou...

technical field

[0001] The invention relates to a method for extracting and preparing an active ingredient in ginkgo leaf extract, in particular to a method for preparing kaempferol-3-O-p-coumaroyl glucorhamnoside from ginkgo leaf extract, which 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, C, J,...

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. One-dimensional liquid chromatography adopts silica gel as the matrix of hydrophilic chromatographic filler 50 × 250mm, 10 μm (Huapu New Innovation Technology Co., Ltd.), and the mobile phase adopts methanol containing 0.1% volume concentration of formic acid as the organic phase, containing 0.1% volume The water with formic acid concentration is the water phase, and the gradient elution method: the concentration of the organic phase decreases from 95% to 90% in step gradient in 0-15min. Use DAD ultraviolet detector 360nm to select the absorption wavelength, the preparation temperature is room temperature, the injection volume is 500μL / needle, the mobile phase flow rate is 90mL / min, the fractions coll...

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 200μL / needle, the flow rate of the mobile phase is 60mL / min, the fractions collected for 15-20 minutes are concentrated to dryness by rotary evaporation , for the one-dimensional preparation of crude kaempferol-3-O-p-coumaroyl glucorhamnoside. Dissol...

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 a hydrophilic chromatographic packing (50×250mm, 10 μ, Huapu Innovation Technology Co., Ltd.) based on silica gel, and the mobile phase adopts ethanol (containing 0.1% formic acid) as the organic phase, water (containing 0.1% % formic acid) is the aqueous phase, and is eluted by 95% organic equivalence. The DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is 30°C, the injection volume is 3000μL / needle, the flow rate of the mobile phase is 120mL / min, the fractions collected for 15-20 minutes are concentrated to dryness by rotary evaporation, and the one-dimensional Preparation of crude kaempferol-3-O-p-cou...

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