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ELISA detection reagent kit suitable for clonazepam relict analysis

An enzyme-linked immunosorbent assay, clonazepam technology, applied in analytical materials, measuring devices, instruments, etc., can solve problems such as the inability to meet the requirements of rapidity, convenience and accuracy, the requirements for testing materials are relatively high, and the research start is late. , to achieve the effect of long storage time, simple pretreatment process and no radioactive pollution

Inactive Publication Date: 2008-12-03
JIANGNAN UNIV
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  • Summary
  • 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 and treatment, which can no longer achieve the fast, convenient and accurate modern detection Require
In recent years, research on immunoassay methods for benzodiazepines has been carried out abroad, but domestic research in this area started relatively late, and most of the detection products still need to rely on foreign imports

Method used

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  • ELISA detection reagent kit suitable for clonazepam relict analysis
  • ELISA detection reagent kit suitable for clonazepam relict analysis
  • ELISA detection reagent kit suitable for clonazepam relict analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1 Kit operation and result calculation:

[0018] After the samples to be tested were pretreated, they were made to volume with PBST for later use. Unpack the vacuum packaging bag and take out the microtiter plate, and equilibrate at room temperature for 5 minutes for later use. Prepare 0ng / mL, 0.15ng / mL, 0.5ng / mL, 1.5ng / mL, 5ng / mL, 15ng / mL, 50ng / mL, 100ng / mL clonazepam standard solution, add 50μL standard sample or process well Put the sample to be tested into each well, make 4 replicates of the standard sample and the sample, add 50 μL of diluted antibody, and incubate at 37°C for 30 minutes; pour out the liquid in the well, wash 5 times with diluted PBST, and remove the microplate Invert and pat dry on absorbent paper; add 100 μL enzyme-labeled goat anti-rabbit secondary antibody diluted 1:1000, incubate at 37°C for 30 minutes; pour out the liquid in the well, wash the plate 5 times with diluted PBST, and pat dry ; Mix liquid A and liquid B at a ratio of ...

Embodiment 2

[0020] Example 2 The formation of clonazepam residue analysis enzyme-linked immunosorbent assay kit:

[0021] In this example, the kit contains the following parts:

[0022] (1) enzyme labels coated with clonazepam antigen;

[0023] (2) Sponge bracket;

[0024] (3) 1mg / mL clonazepam standard substance (Sigma company);

[0025] (4) Clonazepam polyclonal antibody;

[0026] (5) horseradish peroxidase-labeled goat anti-rabbit antibody (Kangcheng Bioengineering Company);

[0027] (6) The concentrated washing liquid formula is: 8g sodium chloride, 0.2g potassium dihydrogen phosphate, 3g disodium hydrogen phosphate, 0.4g potassium chloride, 0.5mL Tween-20 and 20mL distilled water;

[0028] (7) Formula of chromogenic solution A: 0.933g citric acid, 3.68g Na 2 HPO 4 12H 2 O, 18 μL 30% HO 2 o 2 and 100mL ultrapure water;

[0029] (8) Formula of chromogenic solution B: 60 mg of tetramethylbenzidine dissolved in 100 mL of ethylene glycol.

Embodiment 3

[0030] Embodiment 3 shelf life experiment:

[0031] Store the kit at 4°C, take 0, 10, 20, 30, 60, 90, 120, 150 and 180d kits respectively, and use the antigen working concentration of 1.25 μg / mL and antibody working concentration of 0.1 μg / mL as the working concentration Concentration, carry out standard sample detection to determine its detection effect. The results of shelf life determination are shown in Table 1, indicating that IC 50 There is little change, and the kit can be stored at 4°C for more than 6 months.

[0032] Table 1 Kit preservation experiment

[0033]

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Abstract

The invention relates to an enzyme-linked immune detection kit suitable for the analysis of clonazepam residues, and belongs to the technology field of the enzyme-linked immune adsorption analysis kit. The kit comprises an enzyme label plate coated with the envelope antigen of the clonazepam, a sponge support, a clonazepam standard, a clonazepam polyclonal antibody, an enzyme sign second antibody, a concentrating and washing liquid, a colored solution and a reaction stopping solution; the envelope antigen of the clonazepam is the coupling compound of 7-amino clonazepam and an egg-white protein; and the enzyme sign second antibody is a horseradish peroxidase labeled goat anti-rabbit antibody. The kit adopts the clonazepam polyclonal antibody, and can accurately and sensitively detect the clonazepam residues or other structurally similar benzodiazepine residues in urine or tissue; the process of pretreating samples is simple; the time consumption is low; a large quantity of samples can be tested simultaneously; furthermore, the cost for sample detection is lower than that of the traditional instrument detection method; the kit has a long retention time, no radioactive pollution, and practical significance for realizing on-site monitoring of the clonazepam residues of the large quantity of samples.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay kit suitable for clonazepam (CLZ) residue analysis, belonging to the technical field of enzyme-linked immunosorbent assay (ELISA) kits. Background technique [0002] Clonazepam (Clonazepam) is a benzodiazepine sedative hypnotics, the chemical name is: 1,3-dihydro-7-nitro-5-(2-chlorophenyl)-2H-1,4-benzo Diazepine-2-one. Used as an animal feed additive, as a growth promoter, it can make animals sleepy and less active, grow faster and change the meat quality. It is also used for sedation before slaughtering and animal quarantine, or before transferring animals to slaughter, so as to reduce animal tension and reduce animal injury and mortality. Drowsiness, weakness, headache, dizziness, nausea, constipation, and occasional rash, liver damage, bone marrow suppression and other adverse reactions can be seen with these drugs. If such drugs are randomly added to the feed, the accumulated drugs will enter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
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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