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Method for separating cerebroside from sea cucumber by high speed counter current chromatography

A high-speed countercurrent chromatography and cerebroside technology, which is applied in the field of separation and preparation of cerebroside from sea cucumbers by high-speed countercurrent chromatography, can solve the problems of low yield, limited research, and long preparation time of cerebroside

Active Publication Date: 2010-11-24
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually adopt the method separation and preparation of forward and reverse silica gel column chromatography, to remove other lipid components such as phospholipids, triglycerides, too many chromatographic steps make the preparation time of cerebroside long, and loss is bigger, obtains low rate, limiting further research

Method used

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  • Method for separating cerebroside from sea cucumber by high speed counter current chromatography

Examples

Experimental program
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Effect test

Embodiment 1

[0012] Extraction and preparation of cerebroside from Cucumaria frondosa: (1) Preparation of crude extract of cerebroside: Weigh 1 kg of Cucumaria frondosa dry powder, add 4 L of chloroform-methanol with a volume ratio of 1:1 and mix solution, soak overnight at room temperature, filter, repeat this step 5 times, combine the filtrates obtained from each time, concentrate under reduced pressure to a paste, and obtain the crude extract of ginseng cerebrosides, which is set aside. (2) Preparation of the solvent used: put petroleum ether, methanol and water with a volume ratio of 5:4:1 in a separatory funnel, shake vigorously to balance the two phases, separate after standing and layering, and take The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase. The upper and lower phases were ultrasonically degassed for 20 minutes respectively, and cooled for later use. (3) Preparation of sample solution: Accurately weigh 168 mg of the above crud...

Embodiment 2

[0014] Extract and prepare cerebrosides from Haitian melon (Acaudina molpadioides): (1) Preparation of Haitian melon cerebroside crude extract: Weigh 1 kg of Haitian melon dry powder, add 3 L of chloroform-methanol mixture with a volume ratio of 2:1, and store at room temperature Soak overnight, filter, repeat this step 3 times, combine the filtrates obtained from each time, and concentrate under reduced pressure to a paste to obtain the crude extract of guacamole cerebrosides, which is set aside. (2) Preparation of the solvent used: put sherwood oil, methanol and water with a volume ratio of 5.5:3.5:1 in a separatory funnel, shake vigorously to balance the two phases, separate after standing and layering, and take The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase. The upper and lower phases were ultrasonically degassed for 20 minutes respectively, and cooled for later use. (3) Preparation of sample solution: Accurately weigh 10...

Embodiment 3

[0016] Extraction and preparation of cerebrosides from Apostichopus japonicus: (1) Preparation of crude extract of cerebrosides from Apostichopus japonicus: Freeze-dry fresh Apostichopus japonicus and pulverize. Weigh 1 kg of imitation japonicus dry powder, add 3.6L of chloroform-methanol mixture with a volume ratio of 3:1, soak overnight at room temperature, filter, repeat this step 4 times, combine the filtrates obtained each time, concentrate under reduced pressure to a paste, The crude extract of cerebrosides from sea cucumber was obtained and set aside. (2) Preparation of the solvent used: put petroleum ether, methanol and water with a volume ratio of 5:3.5:1 in a separatory funnel, vibrate vigorously to balance the two phases, separate after standing for stratification, and take The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase. The upper and lower phases were ultrasonically degassed for 20 minutes respectively, and cooled...

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Abstract

The invention discloses a method for separating cerebroside from sea cucumber by a high-speed countercurrent chromatography, which is characterized by comprising the following steps of: adding mixed solution of chloroform and methanol into sea cucumber dry powder, soaking at room temperature, and concentrating filter liquor to obtain a cerebroside crude extract; mixing petroleum ether, methanol and water, and respectively performing ultrasonic degasification on the formed stationary phase and mobile phase; dissolving the crude extract by using the mixed solvent, and filtering to obtain samplesolution; and pumping the stationary phase into a chromatographic column of the high-speed countercurrent chromatography, adjusting the rotating speed of a main machine and the flow rate of the mobile phase, pumping in the mobile phase, after two phase equilibria, pumping the sample solution into the chromatographic column through an introduction valve, continuously pumping in the mobile phase, collecting by steps, detecting by using a thin layer chromatography, merging the target object, concentrating, and freeze-drying to obtain the cerebroside. The preparation process has the advantages ofsimpleness, simple and convenient operation, high purification efficiency, short separation time, high stability, and suitability for separation of the cerebroside from various sea cucumber samples and industrial production.

Description

technical field [0001] The invention relates to a method for purifying edible and medicinal active ingredients in sea cucumbers, in particular to a method for separating and preparing cerebrosides from sea cucumbers by high-speed countercurrent chromatography. Background technique [0002] Sea cucumber (Holothurian, sea cucumber) is an animal of the class Holothuroidea in the phylum Echinodermata, and is an important marine food and drug resource. Sea cucumbers contain sea cucumber polysaccharides, sea cucumber saponins, sea cucumber collagen peptides, sea cucumber cerebrosides (cerebrosides from sea cucumber), gangliosides and other biologically active substances, which have the functions of improving immune function, anti-tumor, anti-coagulation, and lowering blood lipids And enhance the brain's ability to learn and remember. Finding and developing new drugs and functional factors from various biologically active substances of sea cucumbers has become an important directi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H15/10C07H1/08
Inventor 徐杰薛长湖李兆杰冯婷玉董平王静凤孙通
Owner OCEAN UNIV OF CHINA
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