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Preparation of Fumonisins b by High Speed ​​Countercurrent Chromatography 1 Standard method

A high-speed countercurrent chromatography and fumonisin technology, which is applied in the field of high-speed countercurrent chromatography to prepare fumonisin B1 standard products, can solve the problems of low toxin content, less than 2,000 yuan, and high cost, so as to reduce the loss rate of toxins and improve product recovery. , The effect of reducing the cost of purification

Active Publication Date: 2021-10-26
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent 201510253888.4 discloses a simultaneous preparation of fumonisin B 1 , B 2 and B 3 The method of the standard product, the method extracts fumonisins from the medium of corn, wheat, rice, etc., the document first uses the cation exchange column to purify the toxin, and then adopts the method of medium-pressure preparative chromatography to refine and purify the toxin, and the purity is higher than 98%. Fumonisin B 1 , B 2 , B 3 Each 10mg, the cost is less than 2000 yuan
However, the toxin content prepared by this method is too low to meet the requirements for a large amount of toxins used in toxin detoxification and degradation studies, toxicology animal experiments, etc. Although the cost of a single experiment of this scheme is low, if a large number of toxin standards are prepared, It needs to be prepared and purified many times, and the cost is still high and consumes a lot of human resources

Method used

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  • Preparation of Fumonisins b by High Speed ​​Countercurrent Chromatography <sub>1</sub> Standard method
  • Preparation of Fumonisins b by High Speed ​​Countercurrent Chromatography <sub>1</sub> Standard method
  • Preparation of Fumonisins b by High Speed ​​Countercurrent Chromatography <sub>1</sub> Standard method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation steps are as follows:

[0033] 1. Preparation of volt horse toxin culture: Fusarium proliferatum 195647 inoculated to PDA medium 25 ° C activation Pressure, inoculated into a triangular bottle equipped with 300 g of rice medium, a total of 300 g of rice medium, 25 ° C, cultured in a triangular bottle, 25 ° C, 15 days, and obtained rice culture in a ventilator. Dry, crush to all particles less than 1 mm, and obtain volt horse toxin culture, spare;

[0034] Determination of the content of Voltamotoxin B1, according to GB5009.240-2016 Determination of voltamin in the first method of food: Immunuspracdition purification - post-column derived high-performance liquid chromatography detection of rice cultivation in the concentration of Vioma B1 Then, according to the formula (the weight (kg) of the concentration (mg / kg) × rice culture in the rice culture (Mg / kg) × rice culture, the weight of the prodeamin B1 is calculated. After testing, the FB1 weight = 368.76...

Embodiment 2

[0055] The preparation steps are as follows:

[0056] 1. Preparation of Voltamotoxin Culture: Fusarium ProLiferatum 195647 inoculated to PDA medium 25 ° C activation In accordance with the amount of inoculation of 10 bacteria disc / bottles, a total of 300 g of rice medium, a total of 3 bottles, 25 ° C, cultured 30D, and rice culture is obtained; rice culture is placed in a fume hood. , Crushing all particles less than 1 mm, obtaining volt horse toxin culture, spare;

[0057] 2. Determination of the content of Voltamotoxin B1, according to GB5009.240-2016 Determination of volts in the first method of food: Immunogene column purification - post-column derived high-performance liquid chromatography detection rice culture in rice cultivation B1 The concentration, then according to the formula (FB1 weight (mg) = Voltaic toxin B1 (mg / kg) × rice culture weight (kg)), the rats B1 is calculated. The weight, after testing, the first embodiment in step 1 of the rice culture in the rice cu...

Embodiment 3

[0071] The preparation steps are as follows:

[0072] 1. Preparation of volt horse toxin culture: Fusarium proliferatum 195647 inoculated to PDA medium 25 ° C activation Pressure, inoculated in a triangular bottle equipped with 300 g of rice medium, a total of 300 g of rice medium, 25 ° C, culture 40D, and rice culture in a ventilator, a total of 300 g of rice medium. Dry, crush to all particles less than 1 mm to obtain vpoderotoxin culture;

[0073] 2. Determination of the content of Voltamotoxin B1, according to GB5009.240-2016 Determination of volts in the first method of food: Immunogene column purification - post-column derived high-performance liquid chromatography detection rice culture in rice cultivation B1 The concentration, then according to the formula (FB1 weight (mg) = Voltaic toxin B1 (mg / kg) × rice culture weight (kg)), the rats B1 is calculated. Weight, after testing, the first embodiment in step 1 of the rice culture is = 327.93 mg;

[0074] 3. Removation of im...

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Abstract

The invention relates to a method for preparing fumonisins B by high-speed countercurrent chromatography 1 The method for a standard product belongs to the technical field of agricultural product safety detection; the method includes sample extraction, rough purification by reverse-phase column chromatography, and purification by high-speed countercurrent chromatography; the preparation of fumonisin B by high-speed countercurrent chromatography in the present invention 1 The standard method is stable and can be applied to fumonisin B in various grain matrices such as rice and wheat 1 The preparation of the method has the advantages of large preparation amount, high recovery rate and low cost, and can prepare more than 250 mg of fumonisin B with a purity greater than 95% in a single time 1 For standard products, the single cost is only about 500 yuan.

Description

Technical field [0001] The present invention relates to a high-speed countercurrent chromatography prepared to Satin B. 1 The method of standard products belongs to the field of agricultural safety testing technology. Background technique [0002] Fumonisins (FB) is a sickle toxin, which is mainly generated by F.Proliferatum and F. Verticillioides. It is the earliest to South African scientist Gelderblom et al. 1992. The year is separated from corn. The vpodumotoxin is a class of diyl compounds with different polyhydrol and a propylene carboxylic acid, there are 11 different kinds, but mainly in FB. 1 , FB 2 , FB 3 Three forms exist, where FB 1 The largest harm is the most distributed. Voltaic toxins will contaminate a variety of food crops, existence in grains and agricultural and sideline products around the world, especially corn. FB in 100 corn foods in Brazil 2007-2010 1 The concentration of 0.126 mg / kg ~ 4.348 mg / kg, the detection rate reached 82% (Food Control, 2012, 2...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C213/10C07C219/06C12P13/00C12R1/77
CPCC07C213/10C12P13/001C07C219/06
Inventor 徐剑宏王刚胡俊强史建荣
Owner JIANGSU ACAD OF AGRI SCI