Clonazepam colloidal gold method detection test paper and preparation method thereof

A technology for clonazepam and detection test paper, which is applied in the field of biological immunoassay determination, can solve the problems of relatively high requirements for test materials, is expensive, cannot be adapted to high-throughput rapid detection, etc., and achieves convenient use, easy production, Economical and fast effect

Inactive Publication Date: 2008-12-03
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of clonazepam at home and abroad mainly includes high performance liquid chromatography (HPLC), gas chromatography (GC), thin layer chromatography (TLC), gas-mass spectrometry (GC-MS), liquid- Mass spectrometry (LC-MS), etc., but these methods not only require expensive instruments and equipment, but also have relatively high requirements for test materials, which require further purification before they can be carried out, and cannot be adapted to high-throughput, on-site rapid detection

Method used

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  • Clonazepam colloidal gold method detection test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Preparation of Clonazepam Test Paper

[0017] (1) Coupling clonazepam with bovine serum protein molecules as clonazepam coupling antigen,

[0018] (2) Immunize with clonazepam conjugated antigen to obtain anti-clonazepam polyclonal antibody;

[0019] (3) Reduce chloroauric acid with trisodium citrate reducing agent to produce 20nm-40nm colloidal gold particles;

[0020] (4) Adjust 3 mL of colloidal gold to pH 9, add dropwise 0.015 mL of anti-clonazepam polyclonal antibody with a protein concentration of 0.18 mg / mL under stirring, and then add 5% bovine serum albumin (BSA) solution after standing for 30 minutes. The final concentration is 1%. After standing for at least 30 minutes, centrifuge at 10,000 rpm for 50 minutes, remove the supernatant and resuspend in 3 mL of 0.002mol / L pH 9.0 borate buffer (containing 5% sucrose), repeat twice, and finally use Dissolve in 0.3mL buffer to obtain a stable anti-clonazepam polyclonal antibody-colloidal gold marker;

[0021]...

Embodiment 2

[0023] Example 2: Test paper requirements

[0024] (1) Compliance rate of negative reference products

[0025] Use phosphate buffered saline (PBS: 0.01mol / L, pH 7.4) to prepare a standard solution of promethazine hydrochloride at a concentration of 10ng / mL for 10 parallel tests. Observe the results with normal or corrected eyesight. The reaction time is observed at 5 min. The result should be negative.

[0026] Use phosphate buffered saline (PBS: 0.01mol / L, pH 7.4) to prepare a 10ng / mL barbiturate standard solution for 10 parallel tests, observe the results with normal or corrected eyesight, and observe the results at 5 minutes of reaction time , Should be negative.

[0027] (2) Compliance rate of positive reference products

[0028] Use phosphate buffered saline (PBS: 0.01mol / L, pH 7.4) to prepare clonazepam standards with concentrations of 25, 50, and 100 ng / mL for parallel detection. Perform 10 parallel experiments for each concentration, and the reaction time is 5 min. Observa...

Embodiment 3

[0035] Example 3: How to use test paper

[0036] (1) Sample preparation

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Abstract

A clonazepam colloidal gold method test paper and a preparation method belong to the testing technology field of the biological immunity method. The test paper is composed of a sample pad, a combined release pad, a cellulose nitrate membrane, a water absorbing pad and a plastic base plate; the sample pad, the combined release pad, the cellulose nitrate membrane and the water absorbing pad are sequentially stuck to the plastic base plate; the combined release pad is coated with an anti-clonazepam multi-resistance colloidal gold label; the cellulose nitrate membrane is sequentially coated with clonazepam coupling antigen and goat anti-rabbit IgG antibody, wherein the clonazepam coupling antigen is taken as the test line and the goat anti-rabbit IgG antibody is taken as the quality control line. The clonazepam colloidal gold method test paper and the preparation method have the characteristics of convenient use, economy, rapidness, simple preparation and low cost.

Description

Technical field [0001] A clonazepam colloidal gold method detection test paper and a preparation method thereof belong to the technical field of biological immune method determination. Background technique [0002] Clonazepam (Nitrazepam) is a benzodiazepine sedative and hypnotic, chemical name: 1,3-dihydro-7-nitro-5-(2-chloro-phenyl)-2H-1,4-benzene And diazepine-2-one. It has been used as an animal feed additive, as a growth promoter, has the effect of making animals sleepy and less moving, growing faster and changing meat quality. It is also used before slaughter and animal quarantine, or before animals are transferred for slaughter, to have a sedative effect to reduce animal stress and reduce animal injury and mortality. Drowsiness is common among these drugs, and adverse reactions such as weakness, headache, dizziness, nausea, constipation, and occasional skin rashes, liver damage, and bone marrow suppression can be seen. If such drugs are added to the feed at will, the accum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N33/558
Inventor 胥传来李秋生刘丽强袁缓彭池方胡拥明
Owner JIANGNAN UNIV
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