Technology method for preparing sea cucumber saponin Holotoxin A1 comparison product by utilizing fresh sea cucumber processing waste liquid

A technology of sea cucumber saponin and processing waste liquid, applied in the directions of steroids, organic chemistry, etc., can solve the problem of no report on the preparation of pure HolotoxinA1, and achieve the effect of high utilization value and simple process

Inactive Publication Date: 2011-05-18
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this paper, sea cucumber saponins were isolated and prepared from sea cucumber waste liquid, but for Holotoxin A 1 The preparation of pure products has not been reported

Method used

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  • Technology method for preparing sea cucumber saponin Holotoxin A1 comparison product by utilizing fresh sea cucumber processing waste liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Collect 2000L of fresh japonicus processing waste liquid, concentrate under reduced pressure for the first time at 65°C, and concentrate to a volume of 200L, with a solid content of 16%, then add 45L of 95% ethanol, and take the supernatant after standing.

[0019] (2) The supernatant passes through the AB-8 type macroporous adsorption resin column with a flow rate of 2 times the column volume. After passing through the column, rinse with 50L of pure water and 20L of 20% ethanol solution; then use 30L of 70% ethanol solution Rinse the macroporous adsorption resin column and collect the eluate from this washing, and concentrate the eluate at 60°C to obtain extract.

[0020] (3) Extraction mixed with sample silica gel powder, adopt methanol: dichloromethane: ethyl acetate: water=2: 2: 4: 1, take the lower layer and carry out silica gel column chromatography, and then utilize methanol: dichloromethane: ethyl acetate : water = 2: 2: 4: 1 is the developing system, take t...

Embodiment 2

[0024] (1) Collect 2000L of fresh japonicus processing waste liquid, concentrate under reduced pressure for the first time at 65°C, and concentrate to a volume of 200L, with a solid content of 13%, then add 70L of 95% ethanol, and take the supernatant after standing still.

[0025] (2) The supernatant passes through the AB-8 type macroporous adsorption resin column with a flow rate of 2 times the column volume. After passing through the column, rinse with 50L of pure water and 20L of 20% ethanol solution; then use 30L of 70% ethanol solution Rinse the macroporous adsorption resin column and collect the eluate from this washing, and concentrate the eluate at 50°C to obtain extract.

[0026] (3) Extraction mixed with sample silica gel powder, using methanol: dichloromethane: ethyl acetate: water = 2: 2: 4: 1 to take the lower layer and carry out silica gel column chromatography, and then use methanol: dichloromethane: ethyl acetate: Water=2:2:4:1 was used as the developing syste...

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Abstract

The invention relates to a technology method for preparing a sea cucumber saponin Holotoxin A1 comparison product by utilizing fresh sea cucumber processing waste liquid. The technology method is characterized by comprising the following steps of: (1) concentrating the fresh sea cucumber processing waste liquid at reduced pressure and carrying out alcohol precipitation, and drawing supernate; (2) leading the supernate to flow through a macropore adsorbent resin column, washing and collecting eluent after absorption until reaching saturation, and concentrating the eluent at reduced pressure to obtain extract; (3) mixing the extract with silica gel powder, and carrying out silicagel column chromatography by using a solution system consisting of methyl alcohol, methylene, ethyl acetate and water; using a system in which the ratio of methyl alcohol to methylene to ethyl acetate to water is 2:2:4:1 as an expanding system, taking a lower layer to carry out thin-layer chromatography (TLC), collecting components with silicagel column chromatography mobility of 0.2-0.3, concentrating the components at reduced pressure, and carrying out vacuum drying on the concentrated components to obtain white powder; and (4) dissolving the white powder with acetonitrile aqueous solution, carrying out ultraviolet absorption by using high performance liquid chromatography (HPLC) at the wavelength of 203nm, carrying out purification by utilizing acetonitrile aqueous solution as a flow phase, collecting main peaks, and carrying out the vacuum drying to obtain the sea cucumber saponin Holotoxin A1 comparison product. The technology method is mainly used for preparing sea cucumber saponin Holotoxin A1.

Description

technical field [0001] The invention relates to a method for preparing sea cucumber saponin Holotoxin A by utilizing fresh sea cucumber processing waste liquid 1 The process method of the reference substance. Background technique [0002] Sea cucumber saponins have important biological activities, such as antifungal, antitumor, anti-inflammatory, antiviral, and immunity-enhancing, etc. In 1976, Japanese scholar Kitagawa isolated sea cucumber (stichopus japonicus shlenka) with significant antifungal activity The saponins, holotoxin A and B, have now become the treatment of athlete's foot and athlete's foot. Since then, the research on sea cucumber saponins has attracted the attention of scholars. [0003] my country's research on sea cucumber saponins began in the 1980s, and more than 100 kinds of sea cucumber saponins with different structures have been isolated and purified. There are many methods for the separation and purification of sea cucumber saponins, including th...

Claims

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Application Information

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IPC IPC(8): C07J71/00
Inventor 袁文鹏刘昌衡刘新张永刚夏雪奎王小军孟秀梅张绵松贾爱荣
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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