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Method and kit for accurately detecting folic acid in blood

A detection kit and kit technology, applied in the field of biotechnology detection, can solve the problems of undetectable, high detection cost, low folic acid, etc., and achieve the effects of easy clinical promotion and application, efficient removal of impurities, and strong specificity

Active Publication Date: 2020-12-15
大连博源医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been reports in the literature on the quantitative analysis of folic acid and its analogs using LC-MS / MS, for example: 1. Lisa Striegel published a method for the quantitative analysis of 5-methyltetrahydrofolate in plasma by isotope dilution mass spectrometry method, although it has extremely high accuracy and precision, but the method only detects 5-methyltetrahydrofolate, and the lower limit of quantitation is as high as 4.51nmol / L (2ng / mL) (Striegel, L., Brandl, B., etal.Quantitation of 5-methyltetraydrofolic acid in plasma for determination of folate status and clinical studies by stable isotope dilution assays.PLOS ONE,2019,14(2)); 2.Susanne H published an isotope dilution UPLC-MS / MS assay for serum Although this method can determine the main folic acid subtypes, the extraction of folic acid from a single sample consumes about 7ml of reagents (Kirsch, S.H., Knapp, J.-P., et al. Quantification of key folate forms in serum using stable-isotope dilution ultra performance liquid chromatography–tandem massspectrometry. Journal of Chromatography B, 2010, 878(1), 68–75). Quantitative analysis methods, but this type of method is still based on basic scientific research, and most of them have the following disadvantages: high detection cost, complicated operation, long time consumption, high quantification limit or inability to detect folic acid and 5-formyltetrahydro folic acid, so it cannot be truly applied in clinical laboratories to achieve accurate detection of folic acid

Method used

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  • Method and kit for accurately detecting folic acid in blood
  • Method and kit for accurately detecting folic acid in blood
  • Method and kit for accurately detecting folic acid in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: A precise detection kit for folic acid in blood

[0037] A precise detection kit for folic acid in blood, which consists of the following table:

[0038]

[0039] Among them, the concentration of each concentration in the calibration solution is as follows:

[0040]

Embodiment 2

[0041] Example 2: A precise detection method for folic acid in serum

[0042] The detection method mainly comprises the following main steps:

[0043] 1. Folic acid extraction process based on kit products:

[0044] A. Mix 200 μL serum sample with 20 μL internal standard mixture, and let it stand for 10 minutes to balance;

[0045] B. The specific adsorption material (n is equal to 6; the particle size of the solid phase extraction material silica gel is 50 μm, and the aperture is ) as a filler, take 100mg and put it in a 1mL column tube, activate the obtained specific solid phase extraction column with 300μL activation solution, equilibrate with 300μL balance solution, add the above serum sample, use 500μL eluent 1 and eluent 2 for rinsing (is it rinsing with 500 μL eluent and 500 μL eluent 2 in sequence), then elute with 200 μL extract, dry the collected eluate with nitrogen at 40°C, and finally use Reconstitute with 100μL equilibration solution.

[0046] 2. Liquid mass...

Embodiment 3

[0057] Example 3: Accuracy and precision of an accurate detection method for folic acid in blood

[0058] The implementation plan for the accuracy and precision evaluation of the kit-based detection method is as follows:

[0059] Use mixed serum samples with low, medium and high concentrations as the samples to be tested, and detect 6 samples of each batch of 3 concentrations of samples, and measure 3 batches. The measurement implementation process is the same as that in Example 2, and the calculations are accurate. Accuracy, within-assay CV and between-assay CV, the results are shown in the table below, showing that the method has excellent accuracy and precision performance.

[0060]

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Abstract

The invention discloses a method and a kit for accurately detecting folic acid in blood. 1) The method for accurately detecting folic acid in blood comprises the steps of 1) adding an internal standard into a blood sample, enriching and purifying folic acid by using a specific solid-phase extraction column, and analyzing the purified liquid by using tandem mass spectrometry, and establishing a calibration curve through an internal standard method for quantification by taking the retention time and the ion pair abundance ratio as a qualitative basis, thereby being accurately detecting folic acid, 5-methyltetrahydrofolic acid and 5-formyl tetrahydrofolic acid in blood. 2) The invention also discloses a kit for accurately detecting folic acid in blood, which comprises a specific solid-phase extraction column, an internal standard mixed solution, a calibration solution, an activation solution, an equilibrium solution, leacheate 1 and leacheate 2, an extraction solution and a quality control sample. According to the invention, accurate quantitative detection of folic acid, 5-methyltetrahydrofolic acid and 5-formyl tetrahydrofolic acid in blood can be realized, and good precision and accuracy are achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to a precise detection method and kit for folic acid in blood. [0002] technical background [0003] Folic acid, also known as pteroyl glutamic acid, or vitamin B9, is an important nutrient for the human body. Folic acid in a broad sense is the general term for folic acid and other related compounds with folic acid biological activity. There are about 100 subtypes, and the main forms are folic acid, dihydrofolate, tetrahydrofolate, 5-methyltetrahydrofolate and 5-methylfolate. Acyl tetrahydrofolate, etc., 5-methyltetrahydrofolate is the main form of folic acid in the blood, accounting for about 82% to 93% of the total, and is the main methyl donor in the human body. [0004] Folic acid is an essential vitamin for cell growth and DNA synthesis in the body. It directly affects the synthesis of nucleic acids and amino acid metabolism, and plays an extremely important role in cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/08G01N30/88
CPCG01N30/08G01N30/88
Inventor 曹云峰刘明莉黄婷赵福荣陈建兴许晶
Owner 大连博源医学检验实验室有限公司
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