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Method for detecting content of homocysteine in blood or urine

A technology of homocysteine ​​and content, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of inability to large-scale application, low accuracy, and high price, and achieve elimination of matrix effects, accurate quantification, and improved recovery. Effect

Active Publication Date: 2014-07-02
郑州和合医学检验实验室有限公司
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Problems solved by technology

At present, the quantitative determination of homocysteine ​​mainly adopts the enzyme immunoassay kit method, but the enzyme immunoassay kit method has the disadvantages of long time, high price, low accuracy, and cannot be applied on a large scale.

Method used

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  • Method for detecting content of homocysteine in blood or urine
  • Method for detecting content of homocysteine in blood or urine
  • Method for detecting content of homocysteine in blood or urine

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Embodiment Construction

[0016] Step 1: Use a pipette to pipette the standard product and internal standard solution, and vortex to mix; at the same time, pipette the sample to be tested and the internal standard solution in parallel and vortex to mix;

[0017] Step 2: Add matrix solution (4% by mass bovine serum albumin solution, normal saline or blank serum solution) to the above mixed solution, and vortex to mix;

[0018] Step 3: Add mercaptoethylamine, mercaptoethanol, dithiothreitol or 120g / L TCEP respectively, and mix thoroughly;

[0019] Step 4: Stand still for 10 minutes;

[0020] Step 5: Add protein removal reagent (10% trichloroacetic acid by mass), vortex and mix well, and centrifuge at high speed;

[0021] Step 6: Take the supernatant and dilute it with deionized water;

[0022] Step 7: Take the supernatant solution, inject it with a liquid phase autosampler, separate it through a liquid chromatography column, analyze it with mass spectrometry, and finally quantitatively analyze the cont...

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Abstract

The invention discloses a method for detecting the content of homocysteine in blood or urine. The method comprises the following steps: 1, respectively mixing a standard substance and a sample to be tested with an internal standard solution, adding a matrix solution, and uniformly mixing; 2, adding mercaptoethanol, dithiothreitol or TCEP (trichloroethyl phosphate), uniformly mixing, and allowing the obtained solutions to stand for 10min; 3, adding trichloroacetic acid with the mass percentage of 10%, centrifuging at a high speed, taking the obtained supernatants, diluting the supernatants by deionized water, carrying out liquid phase-mass spectrometry by respectively allowing the diluted supernatants and a mobile phase to flow through a chromatographic column, and finally determining the content of the homocysteine in the blood or the urine. The method for detecting the content of the homocysteine in the blood or the urine of the invention has the advantages of simplicity, rapidness and sensitivity, can be applied to the screening of a large amount of samples, and makes accurate quantification and strong specificity be realized.

Description

technical field [0001] The invention relates to a method for detecting homocysteine ​​content in blood or urine. Background technique [0002] Homocysteine ​​has been widely accepted as an independent cardiovascular risk indicator, and it is another risk factor besides hyperlipidemia, smoking, and diabetes. In addition to being related to vascular diseases, homocysteine ​​is also related to neural tube defects, preeclampsia, Parkinson's disease, slow fetal growth, chronic renal failure and other diseases, as well as the effects of certain drugs and tumors. At present, the quantitative determination of homocysteine ​​mainly adopts the enzyme immunoassay kit method, but the enzyme immunoassay kit method has the disadvantages of long time, high price, low accuracy, and cannot be applied on a large scale. The liquid phase tandem mass spectrometry detection method we established is a simple, efficient, sensitive and highly accurate quantitative determination method, which provid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/72
Inventor 韩鎏崔学峰倪君君宋彦强李玮
Owner 郑州和合医学检验实验室有限公司
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