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Colloidal gold labeled test strip for rapidly detecting trace fipronil and preparation method of colloidal gold labeled test strip

A technology of trace fipronil and test strips, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as poor timeliness, cumbersome and time-consuming, and inability to conduct on-site real-time monitoring, so as to determine image, display image, and protect consumption. health effects

Inactive Publication Date: 2018-06-29
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for detecting fipronil residues at home and abroad mainly use physical and chemical analysis methods, mainly chromatography, including liquid chromatography-tandem mass spectrometry (LC-MS / MS), gas chromatography (GC), liquid chromatography-tandem Mass spectrometry (LC-MS / MS), gas chromatography-mass spectrometry (GC-MS), etc., the detection of fipronil by these methods requires a series of complicated pretreatment and purification processes, which are tedious and time-consuming, from sample pretreatment to detection It takes at least 2 days for the results, and requires large-scale specialized equipment and professional technicians to operate, which cannot meet the needs of rapid detection, and the timeliness is poor
Although the detection method of fipronil immunoassay kit has high sensitivity and the analysis process is relatively simple, it still takes 2-3 hours for the detection process, and requires the assistance of a microplate reader, so it cannot be monitored in real time on site
At present, the main means of rapid screening of drug residues in test strip detection has unique advantages in the screening of fipronil, and it is a detection technology that needs to be developed first. However, the rapid detection method of fipronil test strips is still blank in China

Method used

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  • Colloidal gold labeled test strip for rapidly detecting trace fipronil and preparation method of colloidal gold labeled test strip
  • Colloidal gold labeled test strip for rapidly detecting trace fipronil and preparation method of colloidal gold labeled test strip
  • Colloidal gold labeled test strip for rapidly detecting trace fipronil and preparation method of colloidal gold labeled test strip

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Experimental program
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Effect test

Embodiment 1

[0035] Such as Figure 1-3 Given, the colloidal gold standard test paper strip of the present invention rapidly detects trace amounts of fipronil, the bottom layer is a supporting layer, the middle layer is an adsorption layer, the protective layer is fixed on the adsorption layer, and the adsorption layer is an adsorption fiber layer, a gold Labeling antibody fiber layer, cellulose film layer and water-absorbing material layer at the handle end, in which the carrier protein coupled with fipronil is sprayed on the cellulose film as a detection line, and there are also goat or rabbit anti-mouse IgG, goat anti Rabbit IgG antibody or SPA printed quality control line.

[0036] The labeled antibody in the gold-labeled antibody fiber layer is colloidal gold-labeled anti-fipronil monoclonal antibody or polyclonal antibody.

[0037] The absorbent fiber layer is made of glass fiber cotton, nylon film, polyvinylidene fluoride film or polyester film; the water-absorbing material layer i...

Embodiment 2

[0043] To make rapid detection trace fipronil colloidal gold-labeled test strips, first prepare the coupling fipronil carrier protein and fipronil gold-labeled antibody, so as to prepare the detection line and gold-labeled antibody fiber layer; secondly prepare goat antibody or rabbit Anti-mouse IgG (or goat anti-rabbit IgG) antibody for preparation of quality control lines.

[0044] The preparation method of the colloidal gold standard test paper strip of the present invention rapidly detects trace amount of fipronil, comprises the following steps:

[0045] 1. Preparation of carrier protein coupled to fipronil (i.e. fipronil carrier protein conjugate)

[0046] (1) The hydrolysis method is used to modify the structure of fipronil. Under the condition of strong alkali, the cyano group (-CN) in fipronil (CNT) is hydrolyzed into carboxyl group (-COOH) to obtain CNT-COOH. Dissolve 20 mg of fipronil standard product in 100 μL of acetone, add the acetone solution of fipronil to 3 m...

experiment example

[0061] Experimental example, sensitivity and specificity assay

[0062] Using fipronil solutions with concentrations of 1ng / mL, 2ng / mL, 4ng / mL, 6ng / mL, 8ng / mL, 10ng / mL, 20ng / mL and 40ng / mL as samples, fipronil colloidal gold standard was used to test Paper strips were tested for fipronil. After 5 minutes, the results showed that at 10ng / mL, 20ng / mL and 40ng / mL, all the test strip detection lines did not develop color, and at 8ng / mL, 90% of the test strip detection lines did not develop color, and at 6ng / mL 50% of the test strips did not develop color at the time of mL, and the test lines of 1ng / mL, 2ng / mL and 4ng / mL all developed color, so it is determined that the visual inspection of fipronil colloidal gold standard test strips is the lowest The detection concentration was 10ng / mL, and the sensitivity was set at 10ng / mL.

[0063] Chlorothalonil, glyphosate, imophos, chlorpyrifos, parathion, malathion and dichlorvos at a concentration of 20 μg / mL were used as samples, and f...

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Abstract

The invention discloses a colloidal gold labeled test strip for rapidly detecting trace fipronil and a preparation method of the colloidal gold labeled test strip. According to the test strip, a bottom layer is a supporting layer, a middle layer is an adsorption layer, a protecting layer is fixed on the adsorption layer, and the adsorption layer comprises an adsorption fiber layer, a gold labeledantibody fiber layer, a cellulose membrane layer and a water absorbing material layer at a handle end sequentially from the test end, wherein a cellulose membrane is coated with fipronil coupled carrier protein to serve as a detection line, and a quality control line printed by goat anti or rabbit anti-mouse IgG, goat anti rabbit IgG antibody or SPA is also arranged. A test indicates that the teststrip has higher specificity and higher sensitivity, limit of detection is 10 ng / mL, and the test strip is free of cross reaction with other pesticides or veterinary drugs.

Description

technical field [0001] The invention relates to a device for rapidly detecting trace amounts of pesticides, in particular to a colloidal gold standard test strip for rapidly detecting trace amounts of fipronil and a preparation method thereof. Background technique [0002] Fipronil, trade name Fipronil, English common name Fipronil (fipronil), chemical name (RS)-5-amino-1-(2,6-dichloro-a,a,a-trifluoro-p -Tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile, the molecular formula is C 8 N 2 Cl 4 , molecular formula C12H4Cl2F6N4OS, molecular weight 437.2, is a phenylpyrazole insecticide developed in 1981 by the French company Rhône-Planck. It has been used since the mid-1990s. It is a broad-spectrum insecticide and veterinary drug widely used in the world. It is mainly used for crop pest control, family pet parasite treatment and control, public health and entertainment venue pest control, etc. Widely used in all aspects. It binds to the γ-aminobutyric acid receptor, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 张改平胡骁飞魏凤仙邓瑞广李青梅姚静静孙亚宁邢广旭杨继飞邢云瑞
Owner HENAN ACAD OF AGRI SCI
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