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A method for preparing kaempferol-3-o-rutinoside from ginkgo biloba extract

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

Active Publication Date: 2015-09-23
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 and dissolve it in 50 mL of 40% methanol-aqueous solution to prepare a Ginkgo biloba extract solution with a concentration of 200 mg / mL, pass through a 0.45 μm microporous membrane, and prepare by one-dimensional liquid chromatography. One-dimensional liquid chromatography uses Xterra C18, the mobile phase uses acetonitrile as the organic phase, and water as the aqueous phase. Gradient elution method: the concentration of the organic phase is increased from 10% to 25% in 20 minutes. Use DAD ultraviolet detector to select the absorption wavelength at 360nm, the preparation temperature is room temperature, the injection volume is 500μL / needle, the mobile phase flow rate is 90mL / min, the fractions are collected for 28-30 minutes, and concentrated to dryness by rotary evaporation, which is a one-dimensional preparation Kaempferol-3-O-rutinoside crude product. Dissolve the crude kaempferol-3-O-rutinoside in 50% methanol-water solution at a ...

Embodiment 2

[0044] Weigh 1 g of Ginkgo biloba extract and dissolve it in 100 mL of 80% methanol-aqueous solution to prepare a solution of Ginkgo biloba extract with a concentration of 10 mg / mL, pass through a 0.45 μm microporous membrane, and prepare by one-dimensional liquid chromatography. One-dimensional liquid chromatography uses Xterra C18, the mobile phase uses methanol as the organic phase, water as the aqueous phase, the concentration of the organic phase is 65% isocratic, the DAD ultraviolet detector is used to select the absorption wavelength at 360nm, the preparation temperature is 40°C, and the sample volume It is 200μL / needle, the flow rate of the mobile phase is 60mL / min, and the fractions of 10-11 minutes are collected and concentrated to dryness by rotary evaporation to prepare crude kaempferol-3-O-rutinoside in one dimension. Dissolve the crude kaempferol-3-O-rutinoside in 40% methanol-water solution at a concentration of 20mg / mL, filter through a microporous membrane, and ...

Embodiment 3

[0046] Weigh 100 g of Ginkgo biloba extract and dissolve it in 200 mL of 50% methanol-aqueous solution to prepare a Ginkgo biloba extract solution with a concentration of 500 mg / mL, pass through a 0.45 μm microporous membrane, and perform one-dimensional liquid chromatography preparation. One-dimensional liquid chromatography adopts Xterra C18, the mobile phase adopts methanol as the organic phase, and water as the aqueous phase, and elution is carried out in a 15% organic phase. Use DAD ultraviolet detector to select the absorption wavelength at 360nm, the preparation temperature is 30℃, the injection volume is 3000μL / needle, the mobile phase flow rate is 120mL / min, and the 22-25 minute fractions are collected and concentrated to dryness by rotary evaporation, which is one-dimensional The crude kaempferol-3-O-rutinoside was prepared. Dissolve the crude kaempferol-3-O-rutinoside in 60% methanol-water solution at a concentration of 80 mg / mL, filter through a microporous membrane...

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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, and specifically relates to a method for preparing kaempferol-3-O-rutinoside from ginkgo biloba extract. Background technique [0002] Kaempferol-3-O-rutinoside is a flavonoid compound, which exists in plants of the Compositae family, such as Safflower, Impatiens and Ginkgo biloba. Kaempferol-3-O-rutinoside has the effect of lowering systolic and diastolic blood pressure, mean arterial blood pressure and heart rate. It also has anti-allergic, analgesic and blood sugar lowering effects. It has great effects on the treatment of cardiovascular and cerebrovascular diseases and diabetes. Application potential. At present, for the preparation of kaempferol-3-O-rutinoside, 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 complicated and the process is complicated. At the same ti...

Claims

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Application Information

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Patent Type & Authority Patents(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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