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Method for detecting content of isosuccinic acid in blood or urine

A methylmalonic acid and urine technology, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of limited diagnostic methods, no detection method for measuring blood samples, etc., and achieves elimination of matrix effects, elimination of systematic errors, The effect of increasing the content

Active Publication Date: 2012-07-04
西安和合医学检验所有限公司
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Problems solved by technology

In my country, current clinical diagnostic methods for such diseases are still very limited. Some scholars have used gas chromatography-mass spectrometry and tandem mass spectrometry to detect and analyze the concentration of MMA in urine, but a simple, efficient and sensitive method suitable for measuring blood samples has not yet been established. Detection method

Method used

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  • Method for detecting content of isosuccinic acid in blood or urine
  • Method for detecting content of isosuccinic acid in blood or urine
  • Method for detecting content of isosuccinic acid in blood or urine

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Embodiment Construction

[0018] 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;

[0019] Step 2: Add matrix solution (4% bovine serum albumin solution or blank serum solution) to the above mixed solution, vortex and mix;

[0020] Step 3: Add a small polar ether with pH<7 for extraction, mix well and centrifuge at high speed;

[0021] Step 4: Take part of the supernatant and dry it with nitrogen on a nitrogen blower;

[0022] Step 5: Add derivatization reagent (saturated acidic alcohol solution), vortex to mix, seal, heat at 65°C for 20min to derivatize;

[0023] Step 6: After the derivatization is completed, heat and dry the solvent again;

[0024] Step 7: Add mobile phase reconstitution solution to redissolve (alcohol aqueous solution or acetonitrile aqueous solution containing 1-5‰ ammonium formate or ammonium acetate, the volu...

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Abstract

The invention discloses a method for detecting the content of isosuccinic acid in blood or urine. The method comprises the following steps of: respectively uniformly mixing a standard sample and a sample to be detected with an internal standard solution, adding a matrix solution and uniformly mixing; adding an extracting reagent and mixing uniformly and then centrifuging; taking supernatant and blow-drying; adding saturated acidic alcohol solvent, mixing uniformly, sealing and heating to 65 DEG C and derivatizing for 20 min; blow-drying the solvent after the derivatization is completed; adding mobile phase redissolving liquid for redissolving and mixing uniformly; carrying out ultrasonic extraction, mixing uniformly and centrifuging; taking supernatant and enabling the supernatant and the mobile phase to pass through a chromatographic column for liquid-mass spectrum analysis so as to finally detect the content of isosuccinic acid in blood or urine. The method can be used for detecting the content of isosuccinic acid simply, conveniently, quickly and accurately, can be applied to mass sample screening and has the advantages of accurate quantification and strong specificity.

Description

technical field [0001] The invention relates to a method for detecting the content of methylmalonic acid in blood or urine. Background technique [0002] Methylmalonic acidemia is one of the most common diseases among congenital abnormalities of organic acid metabolism, which is an autosomal recessive genetic disease. The common clinical symptoms of the disease are lethargy, growth retardation, episodic vomiting, dehydration, dyspnea, hypotonia, and rare symptoms include mental retardation, hepatomegaly, and coma. Since the disease was first reported in 1967, domestic and foreign scholars have used different methods to qualitatively and quantitatively analyze MMA in body fluids of patients in order to provide technical services for clinical diagnosis. The methods used include gas chromatography-mass spectrometry and tandem mass spectrometry. In my country, current clinical diagnostic methods for such diseases are still very limited. Some scholars have used gas chromatograp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 崔学峰倪君君宋彦强
Owner 西安和合医学检验所有限公司
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