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Method for simultaneously detecting homocysteine and metabolism-related substances thereof

A technology for homocysteine ​​and related substances, applied in the field of simultaneous detection of homocysteine ​​and its metabolism-related substances, can solve the problem of low reliability of clinical detection, large blood volume required for detection, long time consumption, etc. problems, to reduce the cost of detection, reliable results, and reduce the effect of matrix effects

Pending Publication Date: 2021-05-25
质谱生物科技有限公司
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Problems solved by technology

The antibodies used in immunoassay detection can also recognize other vitamin derivatives, and the antibody-antigen reaction is usually less than 100%, so the results vary greatly and the specificity is poor, and the reliability of clinical detection is not high, especially for the detection of content lower vitamin
There are the following disadvantages in this way: the detection cost is high, and the operation is complicated, the time spent is long, and the blood volume required for detection is large
Currently there is no effective method for the simultaneous detection of homocysteine, methionine and water-soluble vitamins in serum / plasma

Method used

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  • Method for simultaneously detecting homocysteine and metabolism-related substances thereof
  • Method for simultaneously detecting homocysteine and metabolism-related substances thereof
  • Method for simultaneously detecting homocysteine and metabolism-related substances thereof

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Embodiment

[0034] A method for simultaneously detecting homocysteine ​​and its metabolic related substances, the steps are as follows:

[0035] (1) Sample treatment: Take 120 μL of serum sample, add it to a 2 mL centrifuge tube, add 20 μL of internal standard solution, then add 20 μL of dithiothreitol solution, vortex and incubate for 30 minutes, add 460 μL of methanol to precipitate protein, and vortex After 3 minutes, draw 500 μL of sample mixed solution, filter through a 96-hole protein phospholipid filter plate, collect the filtrate, then blow dry the filtrate with nitrogen, and reconstitute it with ultrapure water after drying. Through this treatment, the precipitated protein can be effectively separated, impurities can be removed, ion interference can be reduced, and the matrix effect can be reduced. At the same time, the analyte can be retained to the greatest extent, and its decomposition or oxidation can be avoided, so as to detect the truest analyte content.

[0036](2) Prepara...

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Abstract

The invention relates to a method for simultaneously detecting homocysteine and metabolism-related substances thereof. The method comprises the following steps: pre-treating a blood sample, analyzing and detecting by adopting liquid chromatography-tandem mass spectrometry, and finally quantifying by adopting an internal standard method. The method can be used for simultaneously detecting the contents of homocysteine and metabolism-related substances (methionine, pyridoxine, folic acid and 5-methyltetrahydrofolic acid) in human serum, and carrying out accurate qualitative and quantitative analysis, and is a detection method which is simple in sample treatment, high in flux and reliable in result.

Description

technical field [0001] The invention belongs to the technical field of combined detection of amino acids and vitamins, and in particular relates to a method for simultaneously detecting homocysteine ​​and its metabolic related substances. Background technique [0002] Homocysteine ​​is a sulfur-containing amino acid, which is mainly derived from the metabolism of methionine ingested in the diet. It is an important intermediate product in the process of methionine metabolism, and it does not participate in protein synthesis. Methionine is one of the essential amino acids that constitute the human body and participates in protein synthesis. If the lack of methionine will lead to blockage of protein synthesis in the body, resulting in damage to the body. The metabolism of homocysteine ​​in cells mainly has two metabolic pathways: one is the heavy methylation of methionine, that is, homocysteine ​​re-synthesizes methionine through the cycle of folic acid; the other is Transsul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/884
Inventor 刘霞王猛米文珍张继钢孙海燕孙翠霞靳海娇时瑞海
Owner 质谱生物科技有限公司
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