Preparation method of high-purity soyasaponin monomers

A soybean saponin, high-purity technology is applied in the field of preparation in the field of biotechnology, and can solve the problems of lack and inability to quantitatively analyze soybean saponins, and achieve the effects of insensitivity to temperature changes, accurate quantitative detection results, and high sensitivity

Inactive Publication Date: 2010-11-24
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The fundamental reason is the lack of standard substances of soybean saponin monomer, w

Method used

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  • Preparation method of high-purity soyasaponin monomers
  • Preparation method of high-purity soyasaponin monomers
  • Preparation method of high-purity soyasaponin monomers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Utilize soybean germ powder to prepare soybean saponin crude extract

[0047] A. Degreasing: Weigh 20 g of soybean germ powder, degrease with petroleum ether (30-60° C.) for 3 hours, take it out and place it in a fume hood to dry naturally.

[0048] B, solvent extraction: 20g defatted germ powder, leaching 4hr with 70% ethanol aqueous solution 200ml by the liquid-solid ratio (mL / g) of 10:1, filter, rotary evaporation reclaims ethanol to obtain filtrate.

[0049] C, solid-phase extraction: the above-mentioned soybean saponin extraction filtrate is added in the adsorption column (Φ30mm * 800mm, resin bed height 550mm) that macroporous adsorption resin (AB-8, Tianjin Nankai University chemical plant) is housed, under room temperature, The flow rate is controlled at 1.5BV / h through the macroporous adsorption resin bed; 5BV of deionized water, at room temperature, the flow rate is controlled at 1.5BV / h to remove sugar and other impurities (the phenol sulfuric acid method...

Embodiment 2

[0064] (1) Utilize soybean germ powder to prepare soybean saponin crude extract

[0065] A, degreasing: same example one.

[0066] B, solvent extraction: 20g defatted germ powder, by 10: 1 liquid-to-solid ratio (mL / g) with 200ml of 70% ethanol aqueous solution supplemented with the ultrasonic wave pretreatment 45min that frequency is 20KHz, filter, rotary evaporation reclaims ethanol and obtains filtrate .

[0067] C. Solid phase extraction: Add the above soybean saponin extraction filtrate into an adsorption column (Φ30mm×800mm, resin bed height 550mm) equipped with a macroporous adsorption resin (Amberlite XAD-2), at room temperature, the flow rate is controlled at 1.5BV / h through the macroporous adsorption resin bed; 5BV of deionized water, at room temperature, the flow rate is controlled at 2.0BV / h to wash away impurities such as sugar (phenol sulfuric acid method to verify whether the sugar is washed); 4BV of ≥ 90% ethanol in Rinse the adsorption column at room tempera...

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Abstract

The invention provides a preparation and detection method of high-purity soyasaponin monomers, comprising the following steps: (1) using soybean germ powder to prepare a soyasaponin crude extract; (2) carrying out fractionation on soyasaponin and soybean isoflavone by using gel chromatography; (3) purifying and separating the monomer soyasaponins by using preparative liquid chromatography; (4) detecting the soyasaponin monomers by using liquid chromatography, an evaporative light scattering detector and a diode array detector; and (5) distinguishing classes of the soyasaponin monomers by using a liquid chromatography-mass spectrometry (LC-MS) method. In the invention, the fractionation on the soyasaponin and the soybean isoflavone is successfully carried out, the purified soyasaponin monomers are prepared and a detection baseline is relatively stable, thereby the soyasaponin is favorable in detection effect; quantitative detection results are more correct by using the diode array detector and the evaporative light scattering detector; and the soyasaponin Aa and Ab in class A, the soyasaponin Ba and Bb in class B and the soyasaponin Be in class E are successfully separated and prepared, the purity is high and peak areas of various samples are up to above 95%.

Description

technical field [0001] The invention relates to a preparation method in the field of biotechnology, in particular to a preparation method of a high-purity soybean saponin monomer that can be used as the basis for the quantitative analysis of soybean saponin and each monomeric saponin and the basis for establishing detection standards. technical background [0002] There are 2% glycosides such as saponins and isoflavones in soybeans, of which saponins account for about 0.5%. Soybean saponins have multiple physiological effects and good pharmacological effects such as anti-cancer, regulating immune function, lowering serum cholesterol levels, preventing and treating cardiovascular diseases, antibacterial, anti-virus, liver protection, weight loss, etc., and can be developed into health food or medicine. An important foaming agent, emulsifier, stabilizer and flavor improver with huge market potential. [0003] Japanese scholars Kitagawa Hoon and Okubo Kazuyoshi divided soybean...

Claims

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Application Information

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IPC IPC(8): C07J63/00C07H15/256C07H1/08G01N30/74G01N30/06
Inventor 赵大云黄玉艾严明霞
Owner SHANGHAI JIAO TONG UNIV
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