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Method for preparing kaempferol-3-O-rutinoside from ginkgo leaf extract

A technology of rutinoside and crude rutinoside, which is applied in the field of medicine, can solve the problems of large consumption of organic solvents, cumbersome operation steps, time-consuming and laborious, etc., and achieve the effects of high degree of instrument automation, abundant resources, and convenient operation

Active Publication Date: 2013-05-22
北京华润高科天然药物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, for the preparation of kaempferol-3-O-rutinoside, the plants are mainly extracted by alcohol, and then separated and purified by multiple column chromatography steps such as macroporous resin or dextran gel. The operation steps are cumbersome and the process is complicated. , at the same time, it needs to consume a large amount of organic solvents, which is time-consuming and labor-intensive
The method of preparing kaempferol-3-O-rutinoside from Ginkgo biloba extract by two-dimensional liquid chromatography-mass spectrometry has not been reported yet

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042]Weigh 10 g of Ginkgo biloba extract, dissolve it in 50 mL of 40% methanol-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 preparation. The one-dimensional liquid chromatography adopts Xterra C18, the mobile phase adopts acetonitrile as the organic phase, water as the aqueous phase, and the gradient elution method: the concentration of the organic phase increases from 10% to 25% in 20 minutes. Adopt 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 collected for 28-30 minutes are concentrated to dryness by rotary evaporation, which is one-dimensional preparation Kaempferol-3-O-rutinoside crude product. The crude kaempferol-3-O-rutinoside was dissolved in 50% methanol-water so...

Embodiment 2

[0044] Weigh 1 g of Ginkgo biloba extract, dissolve it in 100 mL of 80% methanol-water solution to 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. The one-dimensional liquid chromatography adopts Xterra C18, the mobile phase adopts methanol as the organic phase, water as the aqueous phase, the organic phase concentration is 65% isocratic, the DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is 40°C, and the injection volume The flow rate of the mobile phase is 200 μL / needle, the mobile phase is 60 mL / min, the fractions collected for 10-11 minutes are concentrated to dryness by rotary evaporation, and the crude product of kaempferol-3-O-rutinoside is prepared in one dimension. Dissolve the crude kaempferol-3-O-rutinoside in 40% methanol-water solution with a concentration of 20 mg / mL, filter through...

Embodiment 3

[0046] Weigh 100 g of Ginkgo biloba extract and dissolve it in 200 mL of 50% methanol-water solution to obtain a Ginkgo biloba extract solution with a concentration of 500 mg / mL, pass through a 0.45 μm microporous membrane, and prepare by one-dimensional liquid chromatography. The one-dimensional liquid chromatography adopts Xterra C18, the mobile phase adopts methanol as the organic phase, water as the aqueous phase, and adopts 15% organic phase for elution. 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, and the fractions collected for 22-25 minutes are concentrated to dryness by rotary evaporation. Preparation of crude kaempferol-3-O-rutinoside. The crude kaempferol-3-O-rutinoside was dissolved in 60% methanol-water solution with a concentration of 80mg / mL, filtered through a microporous membrane, and prepared by two-...

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Abstract

The invention provides a method for preparing kaempferol-3-O-rutinoside from a ginkgo leaf extract, which comprises the following steps: by using a two-dimensional liquid phase chromatography-mass spectroscopy combination technology and taking methanol-water or acetonitrile-water as a mobile phase and a reversed phase C18 chromatographic column as a one-dimensional preparative chromatographic column, performing component cutting on a ginkgo leaf extract, and collecting a target component which is a kaempferol-3-O-rutinoside crude product; and by taking a hydrophilic TECys as a two-dimensional preparative chromatographic column, performing component cutting on the kaempferol-3-O-rutinoside crude product, collecting, and concentrating through rotary evaporation to obtain the high-purity kaempferol-3-O-rutinoside, wherein the purity can be up to 80% or above. According to the invention, the preparation process is high in repetitiveness and favorable in operability; and meanwhile, ginkgo leaves are abundant in resources and easy to acquire. Thus, the invention meets the requirements for large-scale production and can be used for the preparation of raw materials for a Shuxuening injection.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular to a method for preparing kaempferol-3-O-rutinoside from ginkgo biloba extract. Background technique [0002] Kaempferol-3-O-rutinoside is a flavonoid compound that exists in plants such as Asteraceae, Safflower, Impatiaceae, and Ginkgo biloba. Kaempferol-3-O-rutinoside has the effect of lowering systolic and diastolic blood pressure, mean arterial blood pressure and heart rate, and has anti-allergic, analgesic and blood sugar-lowering effects, and has a great effect on the treatment of cardiovascular and cerebrovascular diseases and diabetes application potential. At present, for the preparation of kaempferol-3-O-rutinoside, the plants are mainly extracted by alcohol, and then separated and purified by multiple column chromatography steps such as macroporous resin or dextran gel. The operation steps are cumbersome and the process is complicated. , At the same time, it needs to c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/07C07H1/08
Inventor 丰加涛付冬梅王海涛冯志琼
Owner 北京华润高科天然药物有限公司
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