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Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography

A high-performance liquid chromatography and aminobutyric acid technology, which is applied in the field of high-performance liquid chromatography analysis of aminobutyric acid in samples, can solve the problems of low detection cost and achieve the effects of low detection cost, simple operation and short running time

Active Publication Date: 2012-07-04
YUEYANG INTEGRATED TRADITIONAL CHINESE & WESTERN MEDICINE HOSPITAL SHANGHAI UNIV OF CHINESE TRADITIONAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report about the HPLC analysis method of GABA in the sample that is simple to operate, low in detection cost, high in sensitivity, and fast and effective.

Method used

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  • Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography
  • Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography
  • Method for detecting aminobutyric acid in sample by high efficiency liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (a) Pre-column derivation:

[0036] Take 500 μl of the sample to be derivatized, add 50 μl 0.05mol / L NaH 2 CO 3 Solution and 200μl 2,4-dinitrofluorobenzene with a mass percentage of 0.2%, mix well, 50°C water bath, react for 40min, cool to room temperature, and dilute to 1mL with methanol-water (V:V=1:1) , to obtain sample solution 1.

[0037] (b) HPLC analysis and UV detection:

[0038] The chromatographic conditions are: chromatographic column: Zorbax SB-C18 column, column length 250mm×inner diameter 4.6mm×particle size 5μm; mobile phase A: a mixture of acetonitrile and water with a volume ratio of 1:1; mobile phase B: 0.05M KH 2 PO 4 Solution, adjust pH to 3.5 with phosphoric acid; flow rate: 1.0 ml / min; UV detection wavelength: 350nm; column temperature: 35°C; injection volume: 30μl; gradient elution program:

[0039] time (min) A / % B / % 0.0 10 90 15.0 80 20 23.0 80 20 23.1 10 90 28.0 10 90

[0040] Take the sample ...

Embodiment 2

[0042] (a) Pre-column derivation:

[0043] Take 500μl of the sample to be derivatized, add 40μl 0.05mol / L NaH 2 CO 3 solution and 300 μl 2,4-dinitrofluorobenzene with a mass percentage of 0.2%, mix well, react in a water bath at 60°C for 30 min, cool to room temperature, and dilute to 1 mL with methanol-water (V:V=1:1) , to obtain sample solution 2.

[0044] (b) HPLC analysis and UV detection:

[0045] The chromatographic conditions are: chromatographic column: Zorbax SB-C18 column, column length 250mm×inner diameter 4.6mm×particle size 6μm; mobile phase A: a mixture of acetonitrile and water with a volume ratio of 1:1; mobile phase B: 0.05M KH 2 PO 4 Solution, adjust pH to 3.7 with phosphoric acid; flow rate: 0.8 ml / min; UV detection wavelength: 350nm; column temperature: 30°C; injection volume: 40 μl; Take the sample solution 2 and inject it into the chromatograph to obtain figure 2In the chromatogram shown, the GABA content calculated according to the area normalizat...

Embodiment 3

[0047] (a) Pre-column derivation:

[0048] Take 500 μl of the sample to be derivatized, add 60 μl 0.05mol / L NaH 2 CO 3 solution and 100 μl of 0.2% by mass percentage of 2,4-dinitrofluorobenzene, mix well, 40°C water bath, react for 50 min, cool to room temperature, and dilute to 1 mL with methanol-water (V:V=1:1) , to obtain sample solution 3.

[0049] (b) HPLC analysis and UV detection:

[0050] The chromatographic conditions are: chromatographic column: Zorbax SB-C18 column, column length 250mm×inner diameter 4.6mm×particle size 4μm; mobile phase A: a mixture of acetonitrile and water with a volume ratio of 1:1; mobile phase B: 0.05M KH 2 PO 4 Solution, adjust pH to 3.3 with phosphoric acid; flow rate: 1.2 ml / min; UV detection wavelength: 350nm; column temperature: 40°C; injection volume: 40 μl; Take the sample liquid three and inject it into the chromatograph to obtain image 3 In the chromatogram shown, the GABA content calculated according to the area normalization ...

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Abstract

The invention relates to a method for detecting aminobutyric acid in a sample by high efficiency liquid chromatography. The method comprises the following steps of: performing derivatization reaction between a to-be-tested sample and NaH2CO3 in the presence of a derivatization reagent 2,4-dinitrofluorobenzene; and performing high efficiency liquid chromatography on the derivatization product under the conditions that the C18 column is adopted, the mobile phase A is a mixture of acetonitrile and water at a volume ratio of 1:1, the mobile phase B is 0.05 M KH2PO4 solution with pH of 3.3-3.7, the flow Velocity is 0.8-1.2 mL / min, the column temperature is 30-40 DEG C, the sample size is 20-40 MuL; and the UV detection wavelength is 350 nm, to obtain the content of aminobutyric acid in the sample. The high efficiency liquid chromatography method is advantageous in short operating time, high baseline stability, high recovery rate and good precision and reproducibility, can effectively detect the content of aminobutyric acid in brain tissues, and can be widely applied to clinical examination and scientific research.

Description

technical field [0001] The invention relates to the technical field of chemical analysis, in particular to a high performance liquid chromatography analysis method for aminobutyric acid in a sample. Background technique [0002] Amino acid neurotransmitters are important substances to regulate the physiological activities of the body. The abnormality of these amino acid neurotransmitters is one of the factors that induce a variety of neurological diseases, such as excitotoxicity, epilepsy, chorea, and Parkinson's disease. [0003] A variety of amino acid neurotransmitters contained in animal brains are mainly divided into two categories according to their functions, one is excitatory amino acids, including aspartic acid and glutamic acid, etc.; the other is inhibitory amino acids, including glycine , aminobutyric acid (γ-aminobutyric acid, GABA) and taurine, etc. Among them, the content of GABA in animal brain tissue is about 0.1-0.6mg / gram tissue. It is an important inhib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
Inventor 程艳梅刘春芳王磊于慧娟朱生樑
Owner YUEYANG INTEGRATED TRADITIONAL CHINESE & WESTERN MEDICINE HOSPITAL SHANGHAI UNIV OF CHINESE TRADITIONAL MEDICINE
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