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A double detection test paper for roxarsine and nifedipine acid

A technology of nitric acid and detection test paper, which is applied in measurement devices, instruments, biological material analysis and other directions to achieve the effects of high sensitivity, simple sample processing method, and saving the number of analysis times.

Active Publication Date: 2021-10-29
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The confirmation method for organic arsine residues has been relatively perfect, but there are few researches on detection methods that can be used for the primary screening of large batches of samples. Flux and other advantages, suitable for high-throughput sample screening, so for the rapid detection of roxarsone and nifedipine residues in animal feed, soil or edible animal tissues, colloidal gold detection test paper is more advantageous

Method used

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  • A double detection test paper for roxarsine and nifedipine acid
  • A double detection test paper for roxarsine and nifedipine acid
  • A double detection test paper for roxarsine and nifedipine acid

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1, Synthesis of Roxarsone and Nifexarsine Artificial Complete Antigen

[0039] 1.1 Molecular modification of roxarsone (ROX) and nifetrol (NIT) hapten

[0040] Weigh 13.1g roxarsone and dissolve it in 100mL NaOH aqueous solution with a concentration of 1mol / L, stir continuously in the ice / water mixture, cool to 0°C; add 30.25g NaOH 2 S 2 o 4Stir, the solution boils, and when the color changes from orange to light yellow, add 12mL concentrated HCl and keep at 0°C until the foaming stops and the product precipitates from the solution. After filtering, collect the precipitate and wash it twice with ice-cold water to obtain a cream color. Solid, namely the crude product of 3-amino-4-hydroxyphenylarsine, was vacuum-dried; 6g crude product was dissolved in 25mL H 2 O and 2 mL of concentrated hydrochloric acid and stirred for 15 min and then filtered, added 25% (w / v) sodium acetate solution to the filtrate until the solution was Congo red, cooled the solution at 4 °...

Embodiment 2

[0050] Embodiment 2, the preparation of monoclonal antibody

[0051] 2.1 Animal immunity

[0052] The immunogen ROX-BSA prepared in Example 1 was added into Freund's complete adjuvant and Freund's incomplete adjuvant to emulsify respectively to make Freund's complete adjuvant immunogen and Freund's incomplete adjuvant immunogen (wherein ROX-BSA and Freund's incomplete adjuvant immunogen were prepared) The volume ratio of Freund's complete adjuvant and Freund's incomplete adjuvant was 1:1). Three 6- to 8-week-old female BALB / c mice (purchased from the Animal Experiment Center of Zhengzhou University School of Medicine) were immunized with ROX-BSA Freund's complete adjuvant immunogen by multi-point injection on the back, 10 μg / mouse ; 21 days and 42 days after the first immunization, BALB / c mice were boosted with ROX-BSA Freund's incomplete adjuvant immunogen with the same method and dose; 3 to 4 days before cell fusion, injected via tail vein BALB / c mice were super-immunized ...

Embodiment 3

[0060] Embodiment 3, the preparation of rabbit anti-mouse IgG

[0061] Immune healthy New Zealand rabbits weighing about 2.0kg with purified mouse IgG, and emulsify the antigen with Freund’s complete adjuvant for the first immunization, inject 50 μg / rat subcutaneously at multiple points, and the interval between each booster immunization is 3 weeks, emulsify the antigen muscle with Freund’s incomplete adjuvant Injection, 2 weeks after the last booster immunization, when the antibody titer of the immune serum was determined to be higher than 1:40 by agar diffusion test (AGP), collect hyperimmune rabbit whole blood, separate the serum, and purify the rabbit anti-small Mouse IgG.

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Abstract

The invention discloses a double detection test paper for roxarsone and nifedipine, the test paper includes anti-roxarsone monoclonal antibody and anti-nifepine acid monoclonal antibody, and can simultaneously detect roxarsone and nifedipine Arsine. In the preparation of the antibody, the present invention adopts roxarsone (ROX) and nifexarsine (NIT) as haptens respectively, and transforms the nitro group into an amino group after reduction, and then couples with the amino group of the carrier protein to develop the roxarsone The artificial antigens of sarsenazo and nifedipine acid, the immunogens ROX‑BSA and NIT‑BSA have better immunogenicity. The obtained two kinds of monoclonal antibodies have the advantages of high specificity, high affinity and high sensitivity, which lay the foundation for the preparation of roxarsone and nifedipine acid dual detection test paper. Compared with the existing high-performance liquid chromatography detection technology, the immunochromatography test paper of the present invention has the advantages of simplicity, speed, sensitivity, accuracy, high throughput, etc., and can realize on-site real-time detection.

Description

technical field [0001] The present invention relates to a double detection test paper for roxarsine and nifedipine, in particular to a method for artificially transforming roxarsone and nifedipine hapten, and a specific monoclonal antibody capable of recognizing roxarsine The invention relates to a specific monoclonal antibody capable of recognizing nitroxarsine, and a prepared colloidal gold immunochromatographic double-linked test paper for synchronous detection of roxarsine and nitroxarsine, which belong to the technical field of veterinary drug residue analysis and immunology. Background technique [0002] Both roxarsone and nifexarsine belong to the organic arsine class of veterinary drugs, which have the basic structure of phenylarsine, have a strong inhibitory and killing effect on a variety of intestinal pathogenic bacteria, can promote the growth of livestock and poultry, and improve feed conversion It can promote the deposition of livestock pigment, and has a good ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/577G01N33/543
CPCG01N33/531G01N33/54306G01N33/577G01N2430/00
Inventor 张改平王爱萍周景明牛艳陈玉梅崔惠芳刘燕凯张静
Owner ZHENGZHOU UNIV
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