Method for extracting folic acid from soybeans

A soybean and folic acid technology, applied in the direction of organic chemistry, color/spectral property measurement, etc., can solve the problems of low sensitivity, difficult folic acid quantification, interference with folic acid determination, etc., and achieve the effect of reducing experimental errors.

Inactive Publication Date: 2016-03-30
JILIN ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

High-performance liquid chromatography can quickly and efficiently separate different folic acid derivatives with a small amount of sample, but the sensitivity of this method is lower than that of microbial methods, and endogenous folic acid will interfere with the determination of folic acid, making it difficult to quantify some forms of folic acid (Zhang et al., 2005; Poo-Prieto et al., 2006)

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  • Method for extracting folic acid from soybeans
  • Method for extracting folic acid from soybeans
  • Method for extracting folic acid from soybeans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 soybean sample preparation

[0032] Soybean seeds were crushed to obtain soybean powder, and 0.2 g of soybean powder was weighed, placed in a 2 mL LAXYGENEP tube, and repeated three times. Add 750μL of folic acid extraction buffer (50mM phosphate buffer, PH=6.5, 1% sodium ascorbate, 0.1% 2-mercaptoethanol) to each sample, incubate at 95°C for 15min and place on ice to cool, then add 5mm steel beads , grind the sample in a tissue grinder (1400 rpm for 5 min). The sample grinding process needs to be protected from light.

Embodiment 2

[0033] Example 2 Soybean folic acid extraction

[0034] Add 8 μL α-amylase and 750 μL folic acid extraction buffer to the sample homogenate to avoid viscousness, place at room temperature for 10 minutes, add 100 μL protease, and incubate at 37°C for 1 hour; boil the above mixture at 100°C for 10 minutes to terminate the enzyme action, and quickly cool on ice for 10 minutes Centrifuge at 14,000 rpm for 10 minutes at 4°C; absorb the supernatant, add 10 μL of rat serum to the supernatant according to the volume of 2%, mix well, and incubate at 37°C for 2 hours; boil at 100°C for 10 minutes, put on ice Cool rapidly for 10 minutes; centrifuge at 4°C and 14,000 rpm for 15 minutes; draw the supernatant and put it in a sterile tube for direct measurement or freeze at -80°C. The sample extraction process needs to be protected from light.

Embodiment 3

[0035] The microbial detection method of embodiment 3 soybean folic acid

[0036] (1) Reagent preparation

[0037] Lactic acid bacteria liquid culture medium: add 4.85g of lactic acid bacteria culture medium to 100mL of water, and sterilize at 121°C for 15min.

[0038] Low folic acid medium: Weigh 4.7g of folic acid medium powder, 0.03g of ascorbic acid, and 0.3mL of folic acid standard stock solution, add water to dissolve to 100mL, and sterilize through a 0.22μm filter membrane for later use.

[0039] Analysis buffer: weigh Na 2 HPO 4 .2H 2 O0.38g, NaH 2 PO 4 Dissolve 0.93g and 1g of ascorbic acid in water to make up to 100mL, pass through a 0.22μm membrane filter to sterilize for later use.

[0040] Folic acid medium: Weigh 9.4g of folic acid medium powder, add water to dissolve to a volume of 100mL, boil for 5min, pass through a 0.22μm membrane filter to sterilize for later use.

[0041] (2) Folic acid standard sample preparation

[0042] Weigh 28 mg of folic acid ...

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Abstract

The invention belongs to the field of preparation of plant extracts, and in particular relates to a method for extracting folic acid from soybean seeds. Human and animals cannot synthesize folic acid per self, and various crops are main resources for obtaining the vitamin by human and animals. A single-enzyme method and a three-enzyme method are commonly used for extraction of folic acid from the crops. The extraction conditions of the three-enzyme method are different in different food, and optimization of folic acid extraction is required to be performed on each kind of food respectively. The invention provides an optimized method for extracting the folic acid from the soybean seeds, by using the method, hundreds of samples can be extracted at the same time, and the extracting process is rapid and effective.

Description

technical field [0001] The invention belongs to the preparation of plant extracts, in particular to a soybean seed extraction and content determination method. Background technique [0002] Folic acid is a water-soluble B vitamin. Folic acid deficiency can cause serious health problems such as neural tube defects, cancer and cardiovascular diseases in infants (Kim et al., 1998; Geisel, 2003; Choi & Friso, 2005; Marisa et al., 2005). Humans and animals cannot synthesize folic acid by themselves, and various crops are the main sources of this vitamin for humans and animals. However, the content of folic acid in crops is usually very low, especially in cereals, how to obtain reliable data on the content of various folic acid derivatives in crops is a key issue in this research field. For the extraction of folic acid from crops, single-enzyme method and three-enzyme method are commonly used. The single-enzyme method is effective for extracting folic acid from foods such as spi...

Claims

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Application Information

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IPC IPC(8): C07D475/04G01N21/31
CPCC07D475/04G01N21/31
Inventor 张玲孟繁磊张原宇王玉民李海云邢国杰董英山
Owner JILIN ACAD OF AGRI SCI
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