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Method for detecting contents of folic acid and sulfur-containing amino acid in plasma

An amino acid and folic acid technology, applied in the field of analytical chemical detection, can solve the problems of low accuracy and high price, and achieve the effect of high accuracy and high accuracy, and solve the inaccurate measurement of cross-reaction

Inactive Publication Date: 2017-02-22
LIAONING RUNSHENG KANGTAI BIOMEDICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Enzyme-linked immunosorbent kit method has relatively low accuracy, high price and can only measure one substance in each determination, so there are certain limitations in its large-scale application

Method used

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  • Method for detecting contents of folic acid and sulfur-containing amino acid in plasma
  • Method for detecting contents of folic acid and sulfur-containing amino acid in plasma
  • Method for detecting contents of folic acid and sulfur-containing amino acid in plasma

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1, Ner reagent solution preparation method

[0047] Use methanol to configure Ner solution containing 2ppmNval, 100mmol / LDTT

[0048] 2. Optimization of mass spectrometry conditions

[0049] After derivatizing folic acid, cysteine, homocysteine, methionine and internal standard norvaline with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, inject directly with a needle pump, In the positive ion mode, optimize the multiple reaction monitoring parameters, in which the ion source temperature is 150°C, the capillary voltage is 3.0kv, the source temperature is 150°C, the desolvation temperature is 500°C, the desolvation gas flow rate is 800L / h, and the cone gas flow rate is 150L / h. The MRM conditions of each compound are as follows:

[0050]

[0051] 3. Optimization of liquid chromatography conditions

[0052] Liquid chromatography mobile phase conditions include A and B: A is an aqueous solution containing 20 mM ammonium formate, 0.5% formic acid, and 1% acetonitrile...

Embodiment 2

[0059] Take 50uL of plasma from the sample to be tested, add 200uL of Ner reagent solution, vortex and mix well, let stand for 10min, and centrifuge at 15000g high speed for 3min at 4°C. Take 10uL of the supernatant and use the Waters AccQ•Tag method for derivatization. The derivatized samples are stored at 4°C and injected uniformly. The processed samples and the mobile phase are separated by a chromatographic column, detected by mass spectrometry, and compared with the standard curve, and finally the contents of folic acid, cysteine, homocysteine ​​and methionine are calculated.

[0060] For the precision test, take matrix plasma and add different concentrations of reference substances, each concentration is 6 samples, according to the sample pretreatment method, add Ner reagent, mix and derivate and analyze, and obtain intra-day and inter-day samples based on the actual measured concentration of each compound. The precision is as follows:

[0061] .

[0062] The method ...

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Abstract

The invention relates to a method for detecting contents of folic acid and sulfur-containing amino acid in plasma and belongs to the technical field of analysis and detection. The method comprises the following steps: firstly adding a reducing agent for reducing cysteine and homocysteine in plasma; secondly, adding a precipitant for performing albumen precipitation; thirdly, taking supernatant, and adding a reaction solvent and a derivatization reagent for carrying out amino acid derivatization reaction; and finally, adopting a chromatographic column for performing separation and mass spectrum detection determination. According to the method provided by the invention, the folic acid, cysteine, homocysteine and methionine in plasma can be determined at one time, and the interference of other matters is eliminated. The method has the advantages of high precision, accuracy and sensitivity and wide linearity range and can be applied to detection of the contents of the folic acid, cysteine, homocysteine and methionine in large quantity of plasma samples.

Description

technical field [0001] The invention relates to an enrichment method for detecting hydrophilic amino acids, which belongs to the technical field of analytical chemistry detection. [0002] technical background [0003] Homocysteine ​​is an intermediate product of methionine cycle and cysteine ​​metabolism in the body, and can be converted into methionine, or converted into cysteine ​​through the transsulfuration pathway. The concentration of homocysteine ​​in human plasma is 5-15umol / L under normal fasting state. Studies have shown that elevated levels of homocysteine ​​in the human body are closely related to cardiovascular and cerebrovascular diseases, peripheral vascular diseases, neurodegenerative diseases, diabetes, pregnancy-induced hypertension, liver cirrhosis, and chronic kidney disease. . In recent years, a large number of studies have reported that folic acid can effectively reduce the concentration of homocysteine ​​in plasma. Therefore, detecting the contents ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/08
CPCG01N30/88G01N30/08
Inventor 孙宏治曹云峰朱志图高鹏葛广波房中则董军洪沫孙晓宇王天一
Owner LIAONING RUNSHENG KANGTAI BIOMEDICAL TECH CO LTD
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