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An electrochemical receptor sensor for detecting β-lactam antibiotics, preparation method and application thereof

A technology of lactams and antibiotics, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of time-consuming, complicated operation, etc., and achieve the effects of high sensitivity, good stability, and wide detection range

Inactive Publication Date: 2020-08-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] To sum up, for the residual β-lactam antibiotics in food, the traditional detection methods have problems such as complicated operation, long time-consuming, and false positive results to varying degrees.

Method used

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  • An electrochemical receptor sensor for detecting β-lactam antibiotics, preparation method and application thereof
  • An electrochemical receptor sensor for detecting β-lactam antibiotics, preparation method and application thereof
  • An electrochemical receptor sensor for detecting β-lactam antibiotics, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of an electrochemical receptor sensor for detecting β-lactam antibiotics

[0037] S1: Preparation of graphene thionine complex.

[0038] Ultrasonic suspend 2mg graphene in 1mL PBS solution, add 2mL 5mg / mL thionine solution and stir at room temperature for 24h; centrifuge at 4000r / min for 5min, wash 3 times with ultrapure water, resuspend 1mg graphene thionine complex in 1mL PBS buffer.

[0039] Characterize the graphene thionine composite prepared above with a UV-Vis spectrophotometer, such as figure 1 It can be seen that the modification was successful, and it was stored at 4°C for future use.

[0040] S2: Induced expression and purification of receptor mutein.

[0041] Induced expression:

[0042] 1. Inoculate a single colony of Escherichia coli pET-28a(+)-BlaR-CTD (known and public) in LB / Kan broth and cultivate for 12h;

[0043] 2. Take 10 mL of bacterial liquid and add it to 1 L of LB broth and 500 μL of kanamycin, place it in a shaker at...

Embodiment 2

[0053] Example 2 Obtaining signal probes

[0054] Preparation of Enzyme Labels

[0055] 1. Weigh 6.18 mg of ampicillin, dissolve it in 1 mL of PBS, pour it into a brown bottle containing magnetic particles, and record it as liquid A;

[0056] 2. Weigh 2.4mg of carbodiimide (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, EDC), dissolve it in 1mL of PBS, add it dropwise to solution A, and weigh N-hydroxysuccinimide (N-Hydroxysuccinimide, NHS) 1.80mg, dissolved in 1mL of PBS, added dropwise to the reaction solution, stirred at room temperature for 2h;

[0057] 3. Weigh 4.3mg of HRP, dissolve it in 2mL of PBS, stir at room temperature until dissolved, and record it as solution B;

[0058] 4. Slowly add solution A to solution B dropwise, and stir for 10 hours at 4°C in the dark;

[0059] 5. Collect the solution, put it into PBS at 4°C for dialysis for 3 days, replace the PBS twice a day, add the same amount of glycerol after the dialysis and freeze at -20°C;

[0060] 6. Measure...

Embodiment 3

[0066] The electrochemical characterization of embodiment 3 working electrode

[0067] In order to monitor the interface properties of electrochemical immunosensors, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used in this study to verify the layer-by-layer modification of the electrodes. The detection conditions of cyclic voltammetry were: voltage -0.2-0.8V, The scanning rate is 50mV / s; the detection conditions of electrochemical impedance spectroscopy: the frequency range is 10-2-105Hz, and the amplitude is 10mV. The electrochemical impedance diagram of the electrode self-assembly process is shown in Figure 7 As shown, the bare GCE electrode shows a very small arc (b scatter point), when the bare GCE electrode is modified with GO / TH, the radius of the arc is significantly reduced and even becomes a straight line (a scatter point), such The results indicated that GO / TH could promote the transfer of [Fe(CN)6]3- / [Fe(CN)6]4- electrons. Howeve...

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Abstract

The invention belongs to the technical field of analytical chemistry and veterinary drug residue analysis, and particularly relates to an electrochemical receptor sensor for detecting beta-lactam antibiotics, and a preparation method and application thereof. The electrochemical receptor sensor comprises an electrochemical probe, a signal probe and a novel composite nano material, the electrochemical probe is thionine, the signal probe is HRP labeled ampicillin (HRP-AMP), and the composite nano material is graphene / thionine / glassy carbon electrode (GO / TH / GCE). The method disclosed by the invention can be used for detecting cephalexin, cefquinome, ceftiofur, penicillin G and ampicillin, thereby providing a rapid and sensitive method for the detection of such substances in foods. The detection method established by the invention has high accuracy and good sensitivity, and has the advantages of saving a lot of time and cost compared with the existing method, and thus has a better market application value.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry and veterinary drug residue analysis, and in particular relates to an electrochemical receptor sensor for detecting β-lactam antibiotics, a preparation method and application thereof. Background technique [0002] β-lactam antibiotics have become the most commonly used antibiotics in the clinical treatment of bacterial infections due to their high efficacy, low price and convenient use, and have also been widely used in the field of animal husbandry, veterinary and aquaculture. At present, it is mainly used to treat cow mastitis, bacterial infection of fish and shrimp and diseases of sea cucumber. However, non-standard medication has serious potential hazards to fishery production and the ecological environment, and it will also lead to drug residues in milk, animals, and aquatic product tissues, causing drug resistance in the human body, resulting in reduced immunity, and endangering...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3276G01N27/48
Inventor 陶燕飞王延新程古月彭大鹏郝海红王旭欧亚红王玉可张莉蕴潘源虎黄玲利王玉莲瞿玮刘振利
Owner HUAZHONG AGRI UNIV
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