Clonazepam colloidal gold method detection test paper and preparation method thereof

A technology for detecting test strips and colloidal gold, which is applied in the field of biological immunoassay determination, can solve problems such as being unable to adapt to high-throughput rapid detection, and achieves the effects of being economical, fast, easy to use and easy to manufacture.

Inactive Publication Date: 2008-12-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of nitrazepam 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 However, these methods not only require expensive instruments and equipment, but also have relatively high requirements for inspection 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 nitrazepam test paper

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

[0018] (2) Immunize with nitrazepam conjugate antigen to obtain anti-nitrazepam polyclonal antibody:

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

[0020] (4) Adjust 3 mL of colloidal gold to pH 9, add dropwise 0.02 mL of anti-nitrazepam polyclonal antibody with a protein concentration of 0.2 mg / mL under stirring, and add 5% bovine serum albumin (BSA) solution to the final product after standing for 30 minutes. The concentration is 1%, after standing for at least 30 minutes, centrifuge at 10,000 rpm for 50 minutes, remove the supernatant and resuspend it in 3 mL of 0.002mol / L pH 9.0 borate buffer (containing 5% sucrose), repeat twice, and finally Dissolve with 0.3mL buffer to obtain stable anti-nitrazepam polyclonal gold marker;

[0021] (5) The anti-ni...

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 with 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 nitrazepam standards with concentrations of 10, 50, and 100 ng / mL for parallel detection. Perform 10 parallel experiments for each concentration and observe the reaction time at 5 min...

Embodiment 3

[0035] Example 3: How to use test paper

[0036] (1) Sample preparation

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Abstract

The invention provides a nitrazepam colloidal gold method detection test paper and a preparation method, belonging to the field of measurement technique of a biological immune method. The detection test paper of the invention consists of a sample pad, a bonded releasing pad, a nitrocellulose film, a water-absorbing pad and a plastic soleplate; the plastic soleplate is sequentially bonded with the sample pad, the bonded releasing pad, the nitrocellulose film and the water-absorbing pad; the bonded releasing pad is wrapped by an anti-nitrazepam multi-resistance-colloid golden marking; the nitrocellulose film is sequentially wrapped by nitrazepam coupling antigen and goat anti-rabbit IgG antibody; the nitrazepam coupling antibody is used as a testing line and the goat anti-rabbit IgG antibody is used as a quality control line. The preparation method of the invention has the advantages of convenience, economical and quick usage, easy fabrication and low cost.

Description

Technical field [0001] A nitrazepam colloidal gold detection test paper and a preparation method thereof belong to the technical field of biological immune method determination. Background technique [0002] Nitrazepam (Nitrazepam) is a benzodiazepine sedative and hypnotic, chemical name: 1,3-dihydro-7-nitro-5-phenyl-2H-1,4-benzodiazepine-2- ketone. Once used as an animal feed additive, as a growth promoter, it 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 with these drugs, and adverse reactions such as weakness, headache, dizziness, nausea, constipation, and occasional skin rash, liver damage, and bone marrow suppression can be seen. If such drugs are added to the feed at will, the accumulated drugs will enter the ...

Claims

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Application Information

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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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