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Method for detecting organic acid/amino acid metabolic product by filter paper shect gas chromatography-mass spectrum analysis

A technology of gas chromatography and mass spectrometry, which is applied in the detection field of organic acids or amino acid metabolites, can solve the problem of easy contamination of gas chromatography columns and mass spectrometry ion sources, insufficient diagnosis and research of complex amino acid diseases, and many miscellaneous peaks in gas chromatography, etc. problems, to achieve the effect of easy computer spectrum library identification, short sample analysis time and high recovery rate

Inactive Publication Date: 2004-09-29
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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Problems solved by technology

However, there are many miscellaneous peaks in gas chromatography, which is not conducive to the analysis of results, and it is easy to contaminate the gas chromatography column and mass spectrometer ion source, and the results of simple urine amino acid analysis are not enough to diagnose and study complex amino acid diseases.

Method used

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  • Method for detecting organic acid/amino acid metabolic product by filter paper shect gas chromatography-mass spectrum analysis
  • Method for detecting organic acid/amino acid metabolic product by filter paper shect gas chromatography-mass spectrum analysis
  • Method for detecting organic acid/amino acid metabolic product by filter paper shect gas chromatography-mass spectrum analysis

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Experimental program
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Embodiment Construction

[0010] 1. Urine specimen collection: Use domestic Xinhua No. 3 filter paper, cut into a size of 5cm×8cm, collect morning urine or any second urine, and put it in a special filter paper drying bag after a little drying.

[0011] 2. Filter paper urine sample processing: dry the filter paper urine sample with 2ml of deionized double-distilled water for multiple elution, centrifuge, and collect the eluate.

[0012] 3. Determine the urine sample concentration: measure the creatinine content to determine the urine concentration, and take a urine sample equivalent to 2.5 μmol creatinine.

[0013] 4. Add 50 μg internal standard pentadecanoic acid and 2-ketocaproic acid to the sample, mix with 100 μl hydroxylamine hydrochloride, 200 μl 0.25M sulfuric acid, and 500 μl saturated ammonium chloride for 45 minutes to complete the oximation reaction of α-keto acid and improve the branched chain Keto acid recovery.

[0014] 5. In an acidic environment, use 3ml ethyl acetate and diethyl ether...

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Abstract

The present invention discloses filter paper sheet gas chromatography-mass spectrum analysis process of organic acid / amino acid metabolic product in urine. The process includes elution of urine collected with filter paper, oximation reaction, extraction, methyl silane derivation, and united detection with chromatographic instrument and mass spectrograph. The process of the present invention is superior in that the filter paper collection of urine favors remote analysis of genetic metabolic diseases; the oximation reaction processing favors alpha-ketonic acid detection for diagnosis of branched chain ketonic acid metabolic disorder; the extraction has high recovery rate of organic acid / amino acid metabolic product, high repeatability, less chromatographic impurity peaks and no pollution to chromatographic capillary column and mass spectrographic ion source; and the united detection is suitable for screening analysis of great amount of samples.

Description

technical field [0001] The invention belongs to a method for detecting organic acid or amino acid metabolites, in particular to a method for detecting urinary organic acid / amino acid metabolites through gas chromatography-mass spectrometry analysis using urine filter paper. Background technique [0002] Because urine contains amino acids, sugars, lipids, and many other endogenous compounds that are terminal or intermediate metabolites, these organic compounds have relatively high water solubility and renal clearance, and their concentrations are usually higher than those in serum and other body fluids, and specimens are the easiest to collect. In the past, my country was only able to carry out preliminary detection of genetic metabolic disease; relying on sending specimens to foreign cooperation units for determination, seriously lagged behind the development of screening, treatment and pathogenesis research of genetic metabolic diseases, affecting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N33/493
Inventor 罗小平王慕逖魏虹梁雁宁琴
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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