Method and kit for simultaneously detecting various metabolites on homocysteine metabolic pathway and application of method and kit

A technology for homocysteine ​​and metabolites, applied in the field of simultaneous detection of multiple metabolites on homocysteine ​​metabolic pathways, can solve the problems of complex serum matrix, affecting the sensitivity and accuracy of results, etc.

Pending Publication Date: 2021-09-03
JINAN YING SHENG BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the complexity of the serum matrix, there are many endogenous substances similar to the physical and chemical properties to be tested, which will affect the sensitivity and accuracy of the results

Method used

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  • Method and kit for simultaneously detecting various metabolites on homocysteine metabolic pathway and application of method and kit
  • Method and kit for simultaneously detecting various metabolites on homocysteine metabolic pathway and application of method and kit
  • Method and kit for simultaneously detecting various metabolites on homocysteine metabolic pathway and application of method and kit

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Embodiment 1

[0072] A method for simultaneously detecting multiple metabolites on the homocysteine ​​metabolic pathway, comprising the steps of:

[0073] Pre-processing method:

[0074] 1) Add 20μg / mLVC to 200uL serum sample, mix well and set aside.

[0075]2) Preparation of calibrator working solution: prepare with physiological saline containing 5% BSA as the matrix, add the mixed standard to the above matrix, so that the final concentrations of Hcy, FA, 5MT, Met, PLP, VB12, Cys, and VC are respectively 50 μmol / L, 20ng / mL, 50ng / mL, 100μmol / L, 80ng / mL, 10ng / mL, 100μmol / L, 20μg / mL. The normal saline containing 5% BSA was used as the diluent to sequentially dilute the above-mentioned mixed standard solution 1:1, and a total of 6 concentration gradients were used as the calibrator working solution. Remarks: VC is used as a protective agent and does not participate in quantification. Take 500 μL of the prepared calibrator working solution of each concentration and divide it into 1.5mL fre...

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Abstract

The invention provides a method and a kit for simultaneously detecting various metabolites on a homocysteine metabolic pathway and application of the method and the kit, and belongs to the technical field of metabolite detection. The method comprises the following steps: detecting a pretreated sample to be detected on the basis of LC-MS / MS; wherein the pretreatment method comprises the step of adding vitamin C into a sample to be detected. By optimizing a pretreatment method, accurate detection of seven substances such as homocysteine, folic acid, 5-methyltetrahydrofolic acid, methionine, vitamin B6, vitamin B12 and cysteine is realized, and the method can be applied to detection items of important metabolites on a clinical homocysteine metabolic pathway. The real level of the real concentration of the metabolite in a subject can be provided, clinical use or laboratory use is facilitated, and the method has important guiding significance on prevention or treatment of hyperhomocysteinemia, so that the method has good practical application value.

Description

technical field [0001] The invention belongs to the technical field of metabolite detection, and in particular relates to a method, a kit and an application for simultaneously detecting multiple metabolites on a homocysteine ​​metabolic pathway. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Homocysteine ​​(Hcy) is a sulfur-containing amino acid, which is mainly produced during the metabolism of methionine (Met). There are three metabolic pathways in the body: ①Homocysteine Under the catalysis of cystathionine β synthase and serine, vitamin B6 is used as a coenzyme to generate cystathionine, and cystathionine is finally generated into cysteine ​​(cysteine, Cys) under the...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 冯振景叶松弭兆元
Owner JINAN YING SHENG BIOTECH
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