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Colloidal gold test strip for detection of imidacloprid and application thereof

The technology of imidacloprid and test strips is applied in the field of detection of imidacloprid residues in tobacco, can solve the problems that it is difficult to meet the rapid detection of a large number of samples and on-site samples, the processing process is complicated, and the time-consuming is long, and the detection time is short, the operation is simple, and the sensitivity is achieved. high effect

Active Publication Date: 2016-06-15
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of these detection methods requires expensive instruments and specialized technicians, and the sample pretreatment process is complex, costly, and time-consuming, and it is difficult to meet the needs of a large number of samples and rapid detection of on-site samples.

Method used

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  • Colloidal gold test strip for detection of imidacloprid and application thereof
  • Colloidal gold test strip for detection of imidacloprid and application thereof
  • Colloidal gold test strip for detection of imidacloprid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of embodiment 1 imidacloprid detection test strip

[0043] The preparation method of this test strip mainly comprises the following steps:

[0044] 1) Prepare a conjugate release pad sprayed with imidacloprid monoclonal antibody-colloidal gold marker;

[0045]2) preparing a reaction membrane with a detection line coated with imidacloprid hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;

[0046] 3) Assemble the conjugate release pad prepared in 1) and 2), the reaction membrane, the sample absorption pad, the water absorption pad and the PVC bottom plate into a test strip.

[0047] The following step-by-step detailed description:

[0048] 1. Synthesis of imidacloprid hapten (see attached for synthetic route figure 1 ) and identification

[0049] 1) Take 1.0g of 2-chloro-5-methylpyridine-3-carboxaldehyde, add carbon tetrachloride to dissolve, add 0.6ml of liquid bromine dropwise at 0°C, stir, slowly...

Embodiment 2

[0086] The detection of imidacloprid residue in the sample of embodiment 2

[0087] 1. Sample pretreatment

[0088] Weigh 1.0±0.05g crushed sample into a 10ml polystyrene centrifuge tube; add 6ml sample extract (80% methanol) and shake for 5min; after standing still, pipette 0.1ml supernatant and 0.9ml sample diluent (0.02 mol / LPBS) to obtain the sample solution to be tested.

[0089] 2. Test with test strips

[0090] Use a pipette to take 100ul of the sample solution to be tested and drop it vertically into the sample hole; start timing when the liquid is flowing, and the result can be interpreted after 5 minutes of reaction, and the interpretation result is valid within 10 minutes.

[0091] 3. Analyze test results

[0092] Negative (-): The color of the T line is darker than that of the C line or consistent with the color of the C line, indicating that the concentration of imidacloprid in the sample is lower than the detection limit, such as image 3 a, 3b.

[0093] Pos...

Embodiment 3

[0095] Example 3 sample detection example

[0096] 1. Detection limit test

[0097] Take a blank tobacco leaf sample, add imidacloprid to it to a final concentration of 2.5 mg / kg, 5 mg / kg, and 10 mg / kg, respectively, take a test strip for detection, and repeat the measurement three times for each sample.

[0098] When testing tobacco samples with test strips, when imidacloprid is added at a concentration of 2.5 mg / kg, the test strip shows that the T line color is darker than the C line color or is consistent with the C line color, and it is negative; When the concentration of imidacloprid is 5mg / kg and 10mg / kg, the test strip shows that the T line color is lighter than the C line color or the T line does not develop color, which is positive, indicating that the test strip can detect imidacloprid in tobacco. The limit is 5mg / kg.

[0099] 2. False positive rate and false negative rate test

[0100] Take 20 tobacco positive samples with a known imidacloprid content greater tha...

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Abstract

The invention provides a colloidal gold test strip for detection of imidacloprid and application thereof. The test strip includes a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate. Specifically, the reaction membrane is provided with a detection line coated with an imidacloprid hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse antibody antibody, the imidacloprid hapten takes 2-chloro-5-methylpyridine-3-formaldehyde as the starting material, bromination reaction is carried out to obtain an intermediate product, then the intermediate product and 2-nitroaminoimidazoline are subjected to nucleophilic substitution reaction, the obtained product is then subjected to condensation with glycine, thus obtaining a carboxyl imidacloprid hapten with a four-carbon chain length spacer arm. The invention also provides a method using the test strip to detect imidacloprid in the sample. The test strip provided by the invention has the advantages of simple operation, high sensitivity, fast detection speed, low cost, and suitability for big-batch sample screening, etc. and can satisfy the requirements of China tobacco supervision department to carry out on-site supervision and detection work.

Description

technical field [0001] The invention relates to a colloidal gold test strip for detecting imidacloprid and an application thereof, in particular to a colloidal gold test strip for detecting imidacloprid, which is particularly suitable for detecting imidacloprid residues in tobacco. Background technique [0002] Imidacloprid (IMI), 1-(6-chloro-3-pyridylmethyl)-N-nitro-2-imidazolidinine, belongs to a new class of neonicotinoid insecticides. IMI is an insect nicotinic acetylcholine receptor (nAChR) inhibitor. It exerts insecticidal effect by preventing insect nerve signal transduction caused by AchR stimulated by IMI. Low toxicity, broad spectrum, systemic absorption, long residual effect and so on. In recent years, imidacloprid has been widely used in crops such as rice, cotton, tea tree, soybean, and tobacco to control agricultural pests such as planthoppers, scale insects, and whitefly, while it is effective against sucking mouthparts such as aphids and leafhoppers in the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/531G01N33/532G01N21/78
CPCG01N21/78G01N33/531G01N33/532G01N33/558G01N33/577
Inventor 冯才伟罗晓琴冯静万宇平王兆芹蒲小容魏丽丽王建霞
Owner BEIJING KWINBON BIOTECH
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