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Method for extracting folic acid from soybean

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

Inactive Publication Date: 2017-03-08
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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)

Method used

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  • Method for extracting folic acid from soybean
  • Method for extracting folic acid from soybean
  • Method for extracting folic acid from soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Soybean sample preparation

[0032] The soybean seeds were crushed to obtain soybean powder, and 0.2 g of soybean powder was weighed and placed in a 2 mL AXYGEN EP tube, 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℃ for 15min and cool on ice, then add 5mm steel balls , Grind the sample in a tissue grinder (rotating speed is 1400rpm, time is 5min). All samples need to be protected from light during grinding.

Embodiment 2

[0033] Example 2 Soybean Folic Acid Extraction

[0034] Add 8μL of α-amylase and 750μL of folic acid extraction buffer to the sample homogenate to avoid stickiness, place at room temperature for 10 minutes, add 100μL of protease, and incubate at 37°C for 1hr; boil the above mixture at 100°C for 10 minutes to stop the enzyme action, and quickly cool on ice for 10 minutes ; Centrifuge for 10 min at 4℃ and 14000rpm; aspirate the supernatant, add 10μL of rat serum to the supernatant according to the volume of 2%, mix well, incubate at 37℃ for 2h; boil and inactivate at 100℃ for 10min, on ice Cool quickly for 10 minutes; centrifuge for 15 minutes at 4°C and 14000 rpm; pipette the supernatant into a sterile tube for direct determination or freeze storage at -80°C. The sample extraction process needs to be protected from light.

Embodiment 3

[0035] Example 3 Microbial detection method of soybean folic acid

[0036] (1) Reagent preparation

[0037] Lactobacillus liquid culture medium: 4.85g lactic acid bacteria culture medium is added to 100mL water, and sterilized at 121°C for 15min.

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

[0039] Analysis buffer: Weigh Na 2 HPO 4 .2H 2 O 0.38g, NaH 2 PO 4 0.93g, 1g ascorbic acid, dissolved in water and dilute to 100mL, sterilized through 0.22μm filter membrane for use.

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

[0041] (2) Preparation of folic acid standard samples

[0042] Weigh 28mg of folic acid in 4mL of 5% K 2 HPO 4 The solution is then...

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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, and specifically relates to a method for extraction and content determination of soybean seeds. 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 disease 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 for humans and animals to obtain this vitamin. 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, there are commonly used single-enzyme and three-enzyme methods. The single-enzyme method is effective for extracting folic acid fr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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