Methamphetamine monoclonal antibody kit having no cross-reactions with ephedrine and pseudoephedrine

A technology of methamphetamine and a test kit is applied in the detection field of illegal drugs, and can solve problems such as affecting case handling work and negative effects.

Active Publication Date: 2009-12-09
SHANGHAI CRIMINAL SCI TECH RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ephedrine and pseudoephedrine are the main ingredients contained in normal therapeutic drugs such as cold medicines and cough medicines. In drug urine tests and military conscription physical examinations, fa...

Method used

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  • Methamphetamine monoclonal antibody kit having no cross-reactions with ephedrine and pseudoephedrine
  • Methamphetamine monoclonal antibody kit having no cross-reactions with ephedrine and pseudoephedrine
  • Methamphetamine monoclonal antibody kit having no cross-reactions with ephedrine and pseudoephedrine

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preparation example Construction

[0076] The preparation of the complete antigen of the present invention comprises the following steps:

[0077] Step (a):

[0078] Step (b): and

[0079] Step (c):

[0080] Specifically, in the step (a) nitration reaction, the methamphetamine is nitrated, that is, a nitro group is inserted on its benzene ring; in the step (b) reduction reaction: the connected nitro group is reduced to Amino group makes it have active chemical properties; in step (c) coupling reaction: adopt homologous bifunctional amino coupling agent that the methamphetamine of activation is coupled with protein couplings such as KLH, BSA or OVA of macromolecule, so just Immunogenicity that makes methamphetamine a complete antigen.

[0081] The nitration and reduction reactions described therein can be carried out by any method known to those skilled in the art under any suitable conditions. For example, the nitration reaction of the present invention can be carried out under the following condition...

Embodiment 1

[0136] Example 1 Activation of methamphetamine and preparation of hapten

[0137] Step (a) Nitrification reaction

[0138] Add 0.52g of methamphetamine raw material and 10mL of concentrated sulfuric acid into the reaction flask, stir to dissolve them, place in an ice-salt bath and cool to -5°C, add concentrated nitric acid (0.25mL) and concentrated A mixed acid solution consisting of sulfuric acid (0.75mL) 1:3. After the addition, react at 0°C for 2 hours, then pour it into 10 g of crushed ice, and react at 0°C for half an hour after the crushed ice melts. g (crude product, which can be directly used in the next step without purification), reaction yield: 53%, and the reaction product is the target object by MNR analysis.

[0139] Step (b) reduction reaction

[0140] Dissolve 0.5g of methyl-[1-methyl-2-(4-nitrophenyl)ethyl]amine in 1.4ml of water, add 0.55ml of concentrated hydrochloric acid under stirring, gradually increase the temperature, and start adding at 30-40°C...

Embodiment 2

[0143] Example 2 Preparation of Methamphetamine-Hemocyanin (KLH) and Methamphetamine-Bovine Serum Albumin (BSA) Complete Antigen

[0144] The p-amino-methamphetamine (p-NH2-Methamphtamine) was coupled with hemocyanin (KLH) or bovine serum albumin (BSA) under the action of EDC.

[0145] Weigh 100mg of p-amino-methamphetamine, dissolve it in 10ml of double distilled water under stirring, then slowly add 400mg of EDC, shake while adding, adjust the pH value to 4.5 with 0.1M hydrochloric acid, at room temperature (20~25 °C) to continue the reaction for 10 minutes.

[0146] Dissolve 100 mg of hemocyanin (KLH) or bovine serum albumin (BSA) in 5 ml of double-distilled water, add it to the p-amino-methamphetamine solution, and continue the reaction at room temperature (20-25° C.) for 2-4 hours. The reaction product was dialyzed against 0.01M phosphate buffer at 4°C.

[0147] Remove unreacted p-amino-methamphetamine and EDC, and replace the buffer solution 3 to 4 times. The obtained...

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Abstract

The invention discloses a hapten and a complete antigen which are used for methamphetamine detection and antibody preparation. The invention also discloses an anti-methamphetamine monoclonal antibody prepared from the complete antigen, and a colloidal gold marked methamphetamine monoclonal antibody immunoassay plate, which are used for detecting the methamphetamine in pharmaceutical products and human specimens of urine samples and the like. Compared with chromatographic processes of HPLC and the like, the immunoassay plate is simple and portable and is easy to carry, can perform field detection, and needs no expensive devices. By using the immunoassay plate to detect the methamphetamine, the entire test can be finished in 3 to 5 minutes, the detection sensitivity can reach 300 ng, and the immunoassay plate has no cross-reactions with 60 common medicaments and poisons, in particular ephedrine and pseudoephedrine.

Description

technical field [0001] The invention relates to the detection of prohibited drugs, in particular to the enzyme-linked immunological detection of methylaniline. Background technique [0002] Methamphetamine, also known as methamphetamine (Methamphetamine), "ice", is a new type of stimulant drug, and the illegal abuse is serious. [0003] As criminal cases related to methamphetamine increase year by year and the country's management of methamphetamine becomes increasingly strict, this field has put forward higher requirements for the detection of methamphetamine and methamphetamine in biological samples of drug abusers. [0004] At present, the detection of methamphetamine is mainly based on chromatographic methods, such as gas chromatography-chromatography (GC-MS) and high-performance liquid chromatography (HPLC). These chromatographic methods have good sensitivity and specificity, but are cumbersome to operate, require expensive instruments and equipment, and take a long ti...

Claims

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Application Information

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IPC IPC(8): C07C211/49C07K14/765C07K14/77C07K14/795C07K16/44C12N5/20G01N33/577G01N33/536
Inventor 曾立波
Owner SHANGHAI CRIMINAL SCI TECH RES INST
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