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Aptamer-based histamine electrochemical sensor, preparation method thereof, and application of same in river crab detection

An aptamer and electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve problems such as complex operation, high detection limit, and low sensitivity

Pending Publication Date: 2021-09-07
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since histamine itself has no specific UV-absorbing groups and fluorescence characteristics, high-performance liquid chromatography generally requires derivatization and complicated pretreatment, the process is cumbersome, and the instrument is expensive, so it is only suitable for laboratory operations; azo reagent color spectrophotometry The sample pretreatment of the method is also relatively complicated, and the detection limit is relatively high, and the sensitivity is low; although the capillary electrophoresis method has the advantages of simple equipment, fast separation speed, and high sensitivity, its operation is complicated, and only capillary zone electrophoresis can be used directly. The determination of biogenic amines does not require a derivatization process; in order to improve the detection sensitivity of the enzyme-linked immunosorbent assay, the double-antibody sandwich method is commonly used, resulting in relatively high detection costs

Method used

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  • Aptamer-based histamine electrochemical sensor, preparation method thereof, and application of same in river crab detection
  • Aptamer-based histamine electrochemical sensor, preparation method thereof, and application of same in river crab detection
  • Aptamer-based histamine electrochemical sensor, preparation method thereof, and application of same in river crab detection

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Experimental program
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Effect test

Embodiment 1

[0026] Materials: ITO, aptamer, TCEP (tris(2-carboxyethyl)phosphine), MCH (6-mercapto-1-hexanol), ligand DNA

[0027] Histamine aptamer:

[0028] 5'-SH-(CH 2 ) 6 -GCCTGTTGTGAGCCTCCTAACATTTCTATGCTGCAGCCAACTTTTCCATACTTCCAGCTTACCATTTATCCATGCTTATTCTTGTCTCCC-3'

[0029] Ligand DNA:

[0030] 5'-SH-(CH 2 ) 6 -GGGAGACAAGAATAAGCATGGATAAATGGTAAGCTGGAAGTATGGAAAAGTTGGCTGCAGCATAGAAATGTTAGGAGGCTCACAACAGGC-3'

[0031] A. Preparation of aptamer / AuNFs / ITO

[0032] ①Preparation of AuNFs / ITO by constant potential electrodeposition: Conductive glass ITO (5×10mm) was ultrasonically cleaned with acetone, ethanol and ultrapure water for 20 minutes to remove possible pollutants on the surface of ITO; 4 Electrodeposition is carried out in an aqueous solution, and N is passed through before deposition 2 Dissolved oxygen was removed for at least 30 minutes, the deposition potential was 0.3 V, and the deposition time was 1200 s, and a flower-shaped nano-gold modified electrode (AuNFs / ITO) was pre...

Embodiment 2

[0039] River crab sample detection (histamine extraction according to the method of GB5009.208-2016):

[0040] Take 100g of the edible part of river crab, mash it fully with a masher, divide it into two parts and put them into clean containers, seal them and mark them, and store them at -20°C. Accurately weigh 10g (accurate to 0.01g) of the evenly ground river crab 2 sample, add 20mL of 10% trichloroacetic acid solution to soak for 2h to 3h, shake for 2min to mix, filter with filter paper, accurately draw 2.0mL of the filtrate into the separatory funnel, and Add sodium hydroxide solution dropwise to adjust the pH between 10 and 12, add 3 mL of n-pentanol and shake for extraction for 5 minutes, let stand to separate layers, and transfer the n-pentanol extract (upper layer) to a 10 mL graduated test tube. Extract three times with n-pentanol, combine the extracts, and dilute to the mark with n-pentanol. Draw 2.0mL of n-pentanol extract into a separatory funnel, add 3mL of hydroc...

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Abstract

The invention discloses an aptamer-based histamine electrochemical sensor, a preparation method thereof, and application of the same in river crab detection. The aptamer-based histamine electrochemical sensor is prepared by the following steps of: firstly, modifying flower-shaped nanogold on the surface of conductive glass based on an electro-deposition method, meanwhile, connecting a sulfhydrylation modified histamine aptamer on the surface of AuNFs / ITO based on an Au-S bonding effect, constructing a histamine electrochemical sensing interface; preparing bismuth vanadate nano flower balls through hydro-thermal synthesis; and on the basis of a sodium borohydride reduction method, introducing gold nanoparticles to the surfaces of the bismuth vanadate nano flower balls, and self-assembling ligand DNA matched with the histamine aptamer to obtain an electrochemical probe DNA / Au-coated BiVO4, namely the aptamer-based histamine electrochemical sensor.

Description

technical field [0001] The invention relates to an electrochemical sensor and its preparation method and application, in particular to an aptamer-based histamine electrochemical sensor, its preparation method and application in river crab detection. Background technique [0002] Histamine is a biogenic amine, one of the autologous active substances. The longer the storage time of aquatic products such as fish, shrimp, and crabs, the higher the histamine content formed by the decarboxylation of histidine in the protein under the action of microorganisms, so the content of histamine is one of the signs to evaluate the freshness of aquatic products one. When the human body ingests excessive histamine, it can cause allergic reactions, which can be life-threatening in severe cases. Therefore, the rapid, high-sensitivity and real-time detection of histamine has important practical significance. [0003] At present, the main methods for detecting histamine include high performanc...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3278
Inventor 黄和江凌徐生盼张幸李昺之
Owner NANJING NORMAL UNIVERSITY
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