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Method for measuring content of total folic acid and derivatives thereof in vegetables synchronously and quantitatively

A technology for quantitative determination of folic acid derivatives, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problems of inability to distinguish folic acid derivatives, cumbersome experimental operations, and different physiological functions of folic acid derivatives.

Inactive Publication Date: 2011-09-07
ZHEJIANG UNIV
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Problems solved by technology

At present, there are many methods for detecting folic acid. The microbial method has the advantages of high sensitivity and accurate results. It is the classic method for the determination of folic acid at present. However, this method has a long cycle and complicated experimental operations, and only the total folic acid data in the sample can be obtained. Cannot distinguish folic acid derivatives
However, because the physiological functions of each folic acid derivative are different, when the research needs to detect the content of some specific folic acid derivative components, this method is not suitable for such research
High-performance liquid chromatography (HPLC) can separate different forms of folic acid derivatives, but at present, most reports use high-performance liquid phase ultraviolet detectors to detect samples with a single composition and high folic acid content, such as the determination of folic acid in folic acid pharmaceuticals, which is not suitable for low folic acid content Determination of Vegetable Samples

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  • Method for measuring content of total folic acid and derivatives thereof in vegetables synchronously and quantitatively
  • Method for measuring content of total folic acid and derivatives thereof in vegetables synchronously and quantitatively
  • Method for measuring content of total folic acid and derivatives thereof in vegetables synchronously and quantitatively

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Embodiment 1

[0060] Embodiment 1. Description of the test case of the present invention to detect vegetable folic acid standard substance BCR-485 (from the Institute of Standard Reference Materials and Methods, Belgium).

[0061] 1. Linear relationship investigation and sensitivity test

[0062] Under the chromatographic conditions of the present invention, select standard folic acid derivative solution respectively for use: tetrahydrofolate (0.5,1,5,15,30,75,100ng / mL), 5-methyl-tetrahydrofolate (0.3,1, 5, 15, 30, 75, 100ng / mL), 5-formyl-tetrahydrofolate (10, 30, 50, 100, 200, 400, 600ng / mL) and folic acid (10, 30, 50, 100, 200 , 400, 600ng / mL), the injection volume was 20 μl, and the peak area integral value was measured, with the injection volume as the abscissa and the peak area as the ordinate, a linear regression equation was obtained, and the regression coefficients are shown in Table 1.

[0063]Standard folic acid derivatives tetrahydrofolate (0.5-100ng / mL), 5-methyl-tetrahydrofola...

Embodiment 2

[0078] Embodiment 2, with regard to the experimental example explanation of total folic acid and derivative content thereof in a spinach sample that the present invention detects the market to buy

[0079] Take a spinach sample purchased in the market, prepare the test solution according to the method of "2.1, 2.2, 2.3", and prepare the standard folic acid derivative product solution at the same time, measure "2.4" according to the above chromatographic conditions, sample injection volume: 20μl, record The peak area of ​​the main peak was obtained from the peak area according to the external standard method to obtain the content of folic acid derivatives (ng / ml). See Table 4 for the qualitative and quantitative analysis results of various forms of folic acid derivatives in spinach according to formula 2.5. The average value of the total folic acid content of a spinach sample in the market is 126.38 μg / 100g. The main folic acid derivatives in spinach are mainly 5-methyl-tetrahy...

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Abstract

The invention discloses a method for measuring the content of total folic acid and derivatives thereof in vegetables synchronously and quantitatively, which comprises the following steps of: 1) extracting folic acid in the vegetables; 2) adding mouse serum conjugase into the obtained vegetable folic acid extract, incubating and centrifugating to obtain vegetable folic acid deconjugation extract; 3) performing solid phase extraction on the vegetable folic acid deconjugation extract and purifying; 4) separating four folic acid derivatives comprising tetrahydrofolic acid, 5-methyl-tetrahydrofolic acid, 5-formyl-tetrahydrofolic acid and the folic acid in the vegetables by using a chromatographic column; and 5) measuring the folic acid quantitatively. By the method, the content of the total folic acid and different folic acid derivatives can be measured synchronously.

Description

technical field [0001] The invention relates to a method for synchronously determining the content of total folic acid and folic acid derivatives in vegetables by a high-performance liquid chromatography-ultraviolet / fluorescence detector. Background technique [0002] Folic acid refers to substances such as tetrahydrofolate and its derivatives. It belongs to the water-soluble B vitamins. As the main one-carbon unit donor, folic acid participates in the biosynthesis of nucleic acids, amino acids, and proteins. Folic acid deficiency in the human body can lead to neonatal neural tube defects, birth defects, mental deterioration, megaloblastic anemia and other diseases, and increase the risk of cardiovascular disease and cancer. Plants and microorganisms can synthesize folic acid by themselves, but humans and other higher mammals lack a complete folic acid synthesis system, so it is necessary to contain a certain amount of folic acid in the diet. For this reason, a series of pre...

Claims

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Application Information

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IPC IPC(8): G01N30/08G01N30/74
Inventor 杨肖娥玛德·加海德·伊斯拉姆·舒哈格韦燕燕
Owner ZHEJIANG UNIV
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