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A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof

A quantitative detection method and technology for quantitative detection, applied in measurement devices, instruments, material analysis by electromagnetic means, etc., can solve the problem of difficult to meet the needs of accurate and high-throughput detection of extremely trace amino acids, and achieve the promotion of standardization and standardization. , the method is simple to operate

Inactive Publication Date: 2013-06-19
刘丽宏 +2
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  • Claims
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AI Technical Summary

Problems solved by technology

Traditional amino acid analysis techniques have been difficult to meet the needs of accurate and high-throughput detection of extremely trace amino acids

Method used

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  • A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof
  • A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof
  • A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0026] 1. Applicable instruments

[0027] Liquid chromatography-tandem mass spectrometer (LC-MS / MS), equipped with electrospray ionization source (ESI).

[0028] 2. Sample requirements

[0029] Plasma samples: collect venous whole blood samples, place them in anticoagulant tubes, mix them upside down 5-6 times immediately after blood collection, centrifuge the plasma within 30 minutes, and store it in a freezer at -20°C until testing.

[0030] Urine samples and cerebrospinal fluid: directly frozen at -20°C after sampling for testing.

[0031] Tissue samples: Biological tissues or ground or homogenized and fully extracted with organic solvents, high-speed centrifugation to take the supernatant, diluted with extraction reagents to a suitable concentration, and stored in a freezer at -20°C for testing.

[0032] Other samples that need to measure free amino acids: After fully dissolved or soaked, fully extract with methanol water (1:1v / v), high-speed centrifuge to take the super...

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Abstract

The invention provides a high-sensitivity and high-throughput kit for simultaneous quantitative detection of 30 amino acids and a preparation method thereof. According to the kit, after extraction treatment of amino acids in a complex sample by using organic solvents, precipitating by using methanol, adding stable isotope-labeled amino acid internal labels, performing derivatization treatment by using hydrochloric acid and n-butanol, drying and redissolving, carrying out reversed-phase chromatography gradient separation by adopting C18 bonded phase silica gel as a stationary phase and acetonitrile-water containing heptafluorobutyric acid and formic acid as a mobile phase, and carrying out quantitative detection by adopting a mass spectrometer. The kit of the invention has the advantages that detection sensitivity can reach 10ng / ml, and all methodological indexes can meet the requirements of quantitative detections of amino acids in clinical examinations, industrial productions and scientific researches.

Description

Technical field [0001] The present invention relates to a kit for simultaneous quantitative detection of 30 amino acids developed based on liquid chromatography-tandem mass spectrometry (LC-MS / MS), more specifically, using liquid chromatography-tandem mass spectrometry to detect 30 amino acids in samples A method for high-sensitivity quantitative detection. [0002] The present invention also relates to the production method and application of the kit of the present invention. Background technique [0003] Amino acid is a general term for a class of organic compounds containing amino and carboxyl groups. There are more than 180 kinds of amino acids found in nature, and more than 30 kinds of amino acids are important in the human body. The emergence, existence and extinction of life are all related to protein, and amino acid is the basic unit of protein, so if the amino acid in the human body changes and is not within the normal range, it will lead to abnormal physiological...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N27/62
Inventor 刘丽宏李鹏飞李丹张绪得陶蓓蓓
Owner 刘丽宏
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