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A kind of method for measuring gamma-aminobutyric acid content by micellar electrokinetic capillary electrophoresis

A technology of capillary electrophoresis and aminobutyric acid, applied in the field of chromatographic analysis, to achieve the effect of reducing the velocity gradient, fast, simple and efficient separation, and less reagent consumption

Active Publication Date: 2018-02-13
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the determination methods of γ-aminobutyric acid use paper chromatography, colorimetry, and high-performance liquid chromatography (HPLC), but whether there are other quantitative detection methods suitable for the quantitative detection of γ-aminobutyric acid remains to be studied

Method used

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  • A kind of method for measuring gamma-aminobutyric acid content by micellar electrokinetic capillary electrophoresis
  • A kind of method for measuring gamma-aminobutyric acid content by micellar electrokinetic capillary electrophoresis
  • A kind of method for measuring gamma-aminobutyric acid content by micellar electrokinetic capillary electrophoresis

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Experimental program
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Effect test

Embodiment 1

[0030] (1) Selection of buffer system

[0031] The buffer system and concentration directly affect the migration and separation of ions. The present invention has investigated the component of each buffer system of capillary separation gamma-aminobutyric acid as table 1, to determine optimal component:

[0032] Table 1

[0033]

[0034]

[0035] by combining Figure 1-8 As shown, because the No. 8 buffer is milky and cannot be electrophoresed, it is omitted Figure 8 ,From figure 1 From the point of view, the peaks separated by the No. 1 system are messy, and the peaks are too low to be effectively separated; figure 2 From the point of view, the No. 2 system has multiple peaks interlaced at 10 minutes, and there are two peaks that cannot be completely separated at 16 minutes; image 3 From the point of view, the No. 3 system has multiple peaks interlaced at 12.5, 14 and 26 minutes; Figure 4 From the point of view, the No. 4 system peaked too quickly, resulting in...

Embodiment 2

[0047] Drawing of γ-aminobutyric acid standard curve: Dilute the 0.01g / ml γ-aminobutyric acid standard and test it to make its standard curve and check its accuracy and reproducibility. For the concentration of 0.2mg / ml, it is determined for 3 consecutive injections According to the analysis conditions of capillary electrophoresis, the optimal injection conditions are: 75 μm injection column, 60 mmol / L borax buffer, 9.5 buffer pH, 20kv pressurized sample injection, absorbed by γ-aminobutyric acid The peak area (y) is plotted against the mass concentration (x) to obtain a standard curve (such as Figure 9 ), its standard curve obtained from the experiment is y=0.000252x, and the detection concentration is 0.083~4.166ug / ml, which is linear, and the lowest concentration that can be detected is 0.08ug / ml.

[0048] The relative standard deviation RSD of the peak area and retention time was also calculated to investigate the repeatability and measurement accuracy of the method. The ...

Embodiment 3

[0050] Sample testing

[0051] Application utilizes internal standard method to detect the GABA content of Portunus trituberculatus different tissue parts, finds GABA content (as table 1):

[0052]

[0053] It can be seen from the above table 1 that the content of GABA in the nerve is extremely high, much higher than that in other parts. GABA has the highest content in the nerve because of its ability to regulate and supply energy to the nerve.

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Abstract

The invention relates to a method for determining content of gamma-aminobutyric acid through a micellar electrokinetic capillary chromatography method, and belongs to the technical field of chromatographic analysis. The method comprises the following steps of selection and pretreatment of a fused silica capillary column, electrophoretic analysis conditions, gamma-aminobutyric acid standard curve drawing, sample derivatization and quantitative analysis of gamma-aminobutyric acid. Compared with the prior art, the method has the following advantages that micellar electrokinetic capillary chromatography (MECC) is adopted, sodium dodecyl sulfonate (SDS) and beta-cyclodextrin serve as micelle, an organic modified isopropanol reagent is added, gamma-aminobutyric acid is rapidly, easily and efficiently separated from other free amino acids, and gamma-aminobutyric can be detected within 30 min.

Description

technical field [0001] The invention belongs to the technical field of chromatographic analysis, in particular to a method for measuring the content of gamma-aminobutyric acid by micellar electrokinetic capillary electrophoresis. Background technique [0002] γ-aminobutyric acid is a very important inhibitory neurotransmitter in the central nervous system. It is a naturally occurring non-protein amino acid with extremely important physiological functions. It can promote the activation of the brain, strengthen the brain and improve intelligence. , anti-epilepsy, promote sleep, beautify skin, delay brain aging function, can supplement human inhibitory neurotransmitters, and has good blood pressure lowering effect. Promote the improvement and protection of kidney function. Inhibit fatty liver and obesity, activate liver function. Supplementing a small amount of γ-aminobutyric acid daily is beneficial to the relief of heart and brain blood pressure, and can also promote the ba...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N27/447
CPCG01N27/447G01N30/02
Inventor 卢少坤王春琳李荣华母昌考宋微微
Owner NINGBO UNIV
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