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Method for detecting salbutamol based on SPR technology

A salbutamol and technical detection technology, applied in the field of chemical analysis, can solve problems affecting sensor sensitivity and detection performance, affecting antigen/antibody binding, etc.

Pending Publication Date: 2021-03-19
INST OF QUALITY STANDARDS & TESTING TECH FOR AGRO PROD OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a linking molecule, the density of carboxyl groups in thiols greatly affects the antigen / antibody binding, which in turn affects the sensitivity and detection performance of the sensor
[0004] MPA, which is usually used as a linker molecule for detecting SPR chips, is monomercaptomonocarboxylic acid. In the process of using MPA as SAMs, the density of its carboxyl groups is extremely high, far exceeding the active sites required for immobilized antigens. , which will inevitably affect the combination of antigen / antibody, thereby affecting the sensitivity and detection performance of the sensor

Method used

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  • Method for detecting salbutamol based on SPR technology
  • Method for detecting salbutamol based on SPR technology
  • Method for detecting salbutamol based on SPR technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 prepares SPR chip

[0032] 1. Experimental method

[0033] 1. MoS 2 Modification of the SPR chip: take 10mg MoS2, sonicate at room temperature (22±2°C) for 4h, and centrifuge at 2000rpm for 20min to remove large particles. The supernatant was centrifuged at 6000rpm for 20min, then washed with N,N-dimethylformamide (DMF) 3 times, using 1mL of DMF each time, the collected precipitate was dried at 50°C, dispersed in dimethylformamide , the concentration of the dispersion was 2 mg·mL-1 and stored at 4 °C. Pipette 200mL of the dispersion onto a clean SPR bare chip and dry in air.

[0034] 2. Modification of SPR chips by AuNPs: firstly, the MoS 2 The modified SPR chip was soaked in 0.01mol / L 1,6-hexanedithiol (HDT) solution for 24h, the surface of the gold film was washed with ethanol and water, and dried with nitrogen; then the SPR chip was placed in a 25nm AuNPs solution overnight, rinsed with deionized water, and dried with nitrogen.

[0035] 3. Modifica...

Embodiment 2

[0041] Salbutamol verification method reliability in the detection sample of embodiment 2

[0042] 1. Sample treatment: add 1g pork to 10mL acetonitrile, extract homogeneously for 1min, take 1mL supernatant in a 5mL centrifuge tube, blow dry with nitrogen, then dissolve with 2mL n-hexane, then add 1mL PBS solution, shake well, and Incubate in a water bath at 85°C for 3 minutes, collect the bottom liquid as the test liquid, filter the liquid to be tested through a 0.22 μm filter membrane, take 250 μL for SPR immunosensor detection, and repeat 3 times for each sample level.

[0043] 2. Methodological validation: mix equal volumes of the optimal concentration of SAL-Ab and different concentrations of SAL standard solutions (5, 10, 20, 30, 40, 60, 80, 100, 150ng / mL) at 37°C Incubate for 30 min, repeat the measurement 3 times for each concentration, record the change of refractive index, and make a standard curve. The result shows that this method has good linear range and detecti...

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Abstract

The invention discloses a method for detecting salbutamol based on an SPR technology. According to the method, mercaptan SAMs with different mercapto groups and carboxyl groups are introduced to adjust the concentration of carboxyl functional groups on a surface; traditional connecting molecules arereplaced with double-mercapto group and double-carboxyl group DMSA; and an SPR signal is enhanced byusing MoS2 and AuNPs, and therefore, the detection sensitivity of an SPR biosensor can be enhanced.

Description

technical field [0001] The invention relates to a method for detecting salbutamol based on SPR technology, which belongs to the field of chemical analysis. Background technique [0002] Surface plasmon resonance technology was initially mainly applied to the interaction between macromolecules through optical refractive index changes, such as the investigation of the affinity between antigen-antibody, receptor-ligand, aptamer-recognizer, but due to its fast, The advantages of label-free and high sensitivity are gradually applied to the field of small molecule detection. Among them, the SPR immunosensor is the most widely used, and its detection mode is mainly competition reaction, that is, the antigen or antibody is immobilized on the surface of the SPR chip through a monolayer, and then the antigen / antibody and the detection target compete for the recognition site on the antibody. With the different concentration of the target, the refractive index of the surface of the SPR...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/59
CPCG01N33/53G01N21/59G01N2021/5903
Inventor 颜朦朦李文杰李慧冬陈子雷张文君毛江胜郭长英丁蕊艳
Owner INST OF QUALITY STANDARDS & TESTING TECH FOR AGRO PROD OF SHANDONG ACADEMY OF AGRI SCI
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