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A kind of ELISA detection method and application of pipemidic acid

A detection method and technology of pipemidic acid, which are applied in measurement devices, instruments, scientific instruments and other directions, can solve the problems of failing to meet the detection requirements of pipemidic acid and high detection limit, and achieve the advantages of reducing loss, simple operation and broad application prospects. Effect

Active Publication Date: 2017-07-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection limit of this method is high, which fails to meet the detection requirements of pipemidic acid

Method used

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  • A kind of ELISA detection method and application of pipemidic acid
  • A kind of ELISA detection method and application of pipemidic acid
  • A kind of ELISA detection method and application of pipemidic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Synthesis and identification of pipemidic acid immunogen

[0066] 1. Dissolve 5.2 mg of pipemidic acid and 20 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) in 1 mL of phosphate buffer with 20 μL of sodium hydroxide solution, which is recorded as solution A; weigh 15 mg bovine serum albumin (BSA) and dissolve it in 1 mL phosphate buffer solution, and record it as solution B; add solution A to solution B, stir at room temperature for 30-40 min, and After the reaction, the solution was put into a dialysis bag, and dialyzed with phosphate buffered saline solution at room temperature for 3 days, during which the dialysate was changed 6 times to obtain the target product pipemidic acid-BSA (immunogen).

[0067] 2. Identification of the immunogen

[0068] The pipemidic acid, BSA, and artificial antigen were scanned under ultraviolet light (200-400 nm), and it was found that the absorption curve of the artificial antigen was higher than that of BSA and pi...

Embodiment 2

[0069] Example 2 Synthesis and identification of pipemidic acid coating agent

[0070] 1. The general method is the same as in Example 1, but ovalbumin (OVA) is used as the carrier protein to obtain the target product pipemidic acid-OVA (enveloping source).

[0071] 2. Identification of the original coating

[0072] The pipemidic acid, OVA, and artificial antigen were scanned under ultraviolet light (200-400 nm), and it was found that the absorption curve of the artificial antigen was higher than that of OVA and pipemidic acid (see attached figure 1 Shown) there is a significant change, it can be confirmed that the original coating was prepared successfully.

Embodiment 3

[0073] Example 3 Synthesis and Identification of Quinolones Coating Progen

[0074] 1. Dissolve 5.2 mg of quinolones (ciprofloxacin, pazufloxacin, gatifloxacin, lomefloxacin, sarafloxacin, and glycofloxacin) in 1 mL of phosphate buffer solution, and Add 15 mg ovalbumin (OVA). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide to the above solution, and stir at room temperature for 20-40 min. Put the reacted solution in a dialysis bag, dialyze with phosphate buffer solution at room temperature for 3 days, change the dialysate 6 times during the period, and obtain the target products ciprofloxacin-OVA, pazufloxacin-OVA, gatifloxacin-OVA, OVA, Lomefloxacin-OVA, Sarafloxacin-OVA, and Glinfloxacin-OVA.

[0075] 2. Identification of the original coating

[0076] The quinolones, OVA, and artificial antigen were scanned under ultraviolet light (200-400 nm), and it was found that the absorption curve of the artificial antigen was higher than that of OVA and pipemidic acid (see attach...

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Abstract

The invention discloses an ELISA detection method for pipemidic acid and application thereof. The ELISA detection method is established by using an ELISA detection system consisting of a pipemidic acid antibody and a coating antigen, wherein the coating antigen is a pipemidic acid coating antigen or a quinolones coating antigen; to be specific, the method is a more sensitive and effective ELISA detection method for pipemidic acid, which is established by taking the pipemidic acid antibody and the coating antigen, namely lomefloxacin-OVA as an optimal combination. The ELISA detection method is efficient, convenient, and high in sensitivity and specificity, the linear range (IC20-IC80) of the detection is 0.9-38.5 ng / mL, the half inhibiting concentration (IC50) is 5.99 ng / mL, and the lowest detection limit is 0.31 ng / mL; and besides, the preprocessing method is simple to operate, the requirements for residue detection can be met, and the method can be applied to quick detection of pipemidic acid illegally added into food, and has wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of food safety. More specifically, it relates to an ELISA detection method and application of pipemidic acid. Background technique [0002] Pipemidic Acid (PPA) is a light yellow or light yellow crystalline powder, odorless and bitter, and is a synthetic second-generation quinolone antibacterial drug. The drug is cyclized by condensation of ethyl 2,4-dichloropyrimidine-5-carboxylate and ethyl β-ethylaminopropionate to 2-chloro-5-hydroxy-7,8-dihydro-8-ethylpyridine [2,3-d] Pyrimidine-6-carboxylate, then brominated, condensed with piperazine, hydrolyzed and neutralized. The molecular formula of pipemidic acid is C 14 h 17 N 5 o 3 , molecular weight: 303.32, and its structural formula is shown in formula (III): [0003] . [0004] Pipemidic acid has a strong antibacterial effect on Gram-negative bacilli such as Pseudomonas aeruginosa, Escherichia coli, and Shigella, and is used to treat urinary tract ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 雷红涛陈佳虹卢蓝蓝孙远明徐振林杨金易沈玉栋王弘肖治理
Owner SOUTH CHINA AGRI UNIV
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