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D-amino acid detection kit and detection method thereof

A detection kit and detection method technology, used in the measurement of color/spectral properties, chemical analysis using catalysis, etc., can solve the problems of high degree of specialization and cost, difficult operation, and not widely used. Inexpensive, small dosage, easy to operate

Inactive Publication Date: 2020-06-12
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Gas chromatography is used to analyze the different distribution coefficients of the components in the gaseous amino acid sample between the two phases. However, the sample needs to be desalted, and the derivatization process also requires anhydrous and high temperature conditions, and it is impossible to measure all amino acids with one chromatographic column. Amino acids, so this method is not widely used in the analysis and detection of amino acids; according to the different distribution coefficients of each component between the two phases, the HPLC method separates and detects them through repeated adsorption-desorption processes. The detection results are accurate, but the degree of specialization The enzymatic method is simple and practical, but has the problem of low sensitivity; while the biosensor detection method is mostly related to electrochemical analysis methods, which often require layer-by-layer assembly of electrode surface materials, which is difficult and time-consuming to operate.

Method used

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  • D-amino acid detection kit and detection method thereof
  • D-amino acid detection kit and detection method thereof
  • D-amino acid detection kit and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Such as figure 1 As shown, the operation steps diagram of the D-amino acid detection kit:

[0032] Step 1: Take 5 μL of the target D-amino acid solution and add it to tube A containing 20 μL of 1.0 μM D-amino acid enzyme solution, and react at 37°C for 45 minutes to obtain the product H of the substrate D-amino acid catalyzed by D-amino acid enzyme 2 o 2 , to collect the solution after the reaction.

[0033] Step 2: Add the solution collected in the first step to tube B containing 0.1mM, 100μL 1,4-benzenediboronic acid solution, and react at room temperature for 30 seconds. 1,4-Benzenediboronic acid can be mixed with the first step Product H 2 o 2 The interaction makes the borate bond of 1,4-benzenediboronic acid oxidized to phenolic group, and the solution after the reaction is collected.

[0034] Step 3: Add the solution collected in the second step reaction into tube C containing 1.0 nM gold nanoparticle solution, react at room temperature for 15 minutes, and co...

Embodiment 2

[0040]Taking D-alanine (D-Ala) as an example below, the detection process of the present invention is described in detail:

[0041] 1) Obtain the absorbance of the standard solution of D-amino acid according to the steps described in Example 1, and obtain a standard curve through the relationship between absorbance and concentration, and obtain the corresponding equation; wherein, the standard solution of D-amino acid is a blank sample, 10 μM ,10 1.5 μM,10 2 μM,10 2.5 μM,10 3 μM,10 3.5 μM,10 4 μM,10 4.5 μM,10 5 μM (the detection concentration range is: 10μM to 10 5 μM. ).

[0042] Collect the corresponding absorbance value by the standard solution of D-amino acid to obtain the spectrogram (see image 3 ), then take the concentration of the target solution as the abscissa, and the corresponding absorbance value as the ordinate, to obtain the standard curve and the corresponding binary equation (see Figure 4 ), that is, the binary equation is Y=-0.2337X+0.1653, X is ...

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Abstract

The invention relates to synthesis of kits, in particular to a D-amino acid detection kit and a detection method thereof. The kit contains D-amino acid enzyme, a 1, 4-benzeneboronic acid solution anda gold nanoparticle solution, wherein the volume ratio is 1: (1-5): (10-20). The kit is simple and easy to operate, can realize green and pollution-free D-amino acid detection, and has potential application value in the research fields of molecular biology, bioanalytical chemistry, marine science, tumor science and the like.

Description

technical field [0001] The invention relates to the synthesis of a kit, in particular to a D-amino acid detection kit and a detection method thereof. Background technique [0002] The corrosion failure of engineering materials caused by microbial corrosion has caused serious economic losses to the national economy and threatened people's personal safety at all times [1]. Studies have shown that corrosive microorganisms in the marine environment can synthesize and release signal molecules D-amino acids to the surrounding environment during growth and metabolism. Plays an important regulatory role in membrane disintegration. In addition, the formation, development, and disintegration of biofilms during corrosion are also strictly regulated by microorganisms, and this regulation is often achieved through the response of signal molecules [2]. Therefore, the analysis and research of signal molecules is particularly important in order to gain a deeper understanding of the change...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N31/10G01N21/31
CPCG01N31/10G01N21/31
Inventor 曾艳张盾
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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