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Colorimetric detection probe for detecting kanamycin, and detection method thereof

A technology for kanamycin and detection probes, applied in the field of colorimetric detection probes for kanamycin residues, can solve problems such as enzyme activity interference, achieve low detection limits, good specificity and selectivity, and good application and development prospects

Active Publication Date: 2017-11-24
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the common colorimetric method uses horseradish peroxidase to catalyze the color development of some substrates to achieve target detection, but the activity of the enzyme is very susceptible to interference from external conditions.

Method used

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  • Colorimetric detection probe for detecting kanamycin, and detection method thereof
  • Colorimetric detection probe for detecting kanamycin, and detection method thereof
  • Colorimetric detection probe for detecting kanamycin, and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] like figure 1 As shown, this embodiment includes the following steps:

[0030] (1) Preparation of MOF-Pt composite material: first synthesize Fe-MIL-88, weigh 0.118g terephthalic acid and 0.187g FeCl respectively 3 . 6H 2 O was dissolved in 15mL of DMF, then 3.45mmol of acetic acid solution was added, the oil bath was reacted at 110°C for 3h, and the precipitate was collected after cooling to room temperature; to synthesize functionalized Pt NPs, 16.6mg of PVP was dissolved in 45mL of ethanol, and then 5mL of chloroplatinic acid was added dropwise Added, then stirred at room temperature for 2min, and finally refluxed for 3h to collect the precipitate to obtain Pt NPs-PVP; finally, Pt NPs-PVP was added dropwise to Fe-MIL-88 solution under vigorous stirring, Pt NPs-PVP and Fe -The ratio of MIL-88 is 1:2, react at room temperature for 2 hours, and finally centrifuge to collect the precipitate to obtain MOF-Pt composite material.

[0031] (2) Preparation of composite pr...

Embodiment 2

[0043] This embodiment includes the following steps:

[0044] (1) Preparation of MOF-Pt composite material: first synthesize Fe-MIL-88, weigh 0.354g terephthalic acid and 0.561g FeCl respectively 3 . 6H 2 O was dissolved in 45mL of DMF, then 7.90mmol of acetic acid solution was added, the oil bath was reacted at 120°C for 4h, and the precipitate was collected after cooling to room temperature; to synthesize Pt-PVP, 33.2mg of PVP was dissolved in 60mL of ethanol, and then 8mL of chloroplatinic acid was added dropwise. Then the reaction was stirred at room temperature for 5 min, and finally refluxed for 5 h to collect the precipitated Pt NPs-PVP; finally, the Pt NPs-PVP was added dropwise to the Fe-MIL-88 solution under vigorous stirring, and the Pt NPs-PVP and Fe-MIL The ratio of -88 was 4:1, reacted at room temperature for 6 hours, and finally collected the precipitate by centrifugation to obtain the MOF-Pt composite material.

[0045] (2) Preparation of composite probe sol...

Embodiment 3

[0058] This embodiment includes the following steps:

[0059] (1) Preparation of MOF-Pt composite material: first synthesize Fe-MIL-88, weigh 0.253g terephthalic acid and 0.341g FeCl 3 . 6H 2 O was dissolved in 30mL of DMF, then 5.00mmol of acetic acid solution was added, the oil bath was reacted at 115°C for 4h, and the precipitate was collected after cooling to room temperature; to synthesize Pt-PVP, 22.2mg of PVP was dissolved in 50mL of ethanol, and then 8mL of chloroplatinic acid was added dropwise. Then stir the reaction at room temperature for 3min, and finally reflux for 4h to collect the precipitate to obtain Pt NPs-PVP; finally, add Pt NPs-PVP dropwise to the Fe-MIL-88 solution under vigorous stirring, Pt NPs-PVP and Fe-MIL The ratio of -88 was 2:1, reacted at room temperature for 5 hours, and finally collected the precipitate by centrifugation to obtain the MOF-Pt composite material.

[0060] (2) Preparation of composite probe solution: Apt and MB were combined b...

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Abstract

The present invention relates to a colorimetric detection probe for detecting kanamycin. The preparation method comprises: (1) conjugating an aminated aptamer Apt and aminated magnetic beads MB under the action of a coupling agent to form a capture probe; (2) modifying the surface of a synthesized metal organic framework MOF-Pt composite material by using single-stranded DNA binding protein SSB to form a signal probe; and (3) forming a composite probe from the capture probe obtained in the step (1) and the signal probe obtained in the step (2) through the affinity between the single stranded DNA binding protein and the aptamer Apt. The invention further provides a kanamycin detection method of the probe. According to the present invention, the detection method does not require the large-size instrument equipment and has the low detection limit, the colorimetric detection probe shows good specificity and good selectivity, the operation of the method is simple and quick, the on-site detection of the kanamycin residue can be achieved by using the handheld type colorimeter, and the experimental results are visible to the naked eye.

Description

technical field [0001] The invention relates to the fields of environmental science, laboratory chemistry and analytical chemistry, in particular to a colorimetric detection probe for detecting kanamycin residues in milk based on an enzyme-free catalyzed colorimetric method and a detection method thereof. Background technique [0002] Kanamycin (KANA), as a small-molecule drug, is sensitive to intestinal infection caused by bacteria and used as preparation before intestinal surgery, and has the effect of reducing ammonia produced by intestinal bacteria. Hepatic coma has a certain preventive effect. However, when KANA in the human body accumulates to a certain extent, certain side effects will occur, such as erythema rash, hearing loss, tinnitus or ear fullness (ototoxicity), and nephrotoxicity, etc., and the long-term accumulation of KANA in the body will cause People develop drug resistance, which poses a huge threat to human health. [0003] At present, many methods for ...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/31G01N21/78G01N2021/7756
Inventor 干宁栾倩周游
Owner NINGBO UNIV
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