Magnetic particle chemiluminescence kit for detecting chlorpromazine, and applications thereof

A chemiluminescence reagent and kit technology, used in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problems of poor reagent stability, reaction time and temperature effects, etc. The effect of low time, fast detection and high sensitivity

Inactive Publication Date: 2013-02-13
BEIJING KWINBON BIOTECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The reaction time and temperature have a great influence on the enzyme activity, and the stability of the reagent is poor

Method used

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  • Magnetic particle chemiluminescence kit for detecting chlorpromazine, and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation of the concrete component of embodiment one kit

[0036] 1) Chlorpromazine artificial antigen synthesis.

[0037] Chlorpromazine and bovine serum albumin (BSA) were coupled to the immunogen by the mixed anhydride method.

[0038] 2) Preparation of monoclonal antibodies

[0039] Animal immunization: Immunize Balb / c mice with the immunogen at a dose of 100 μg / mouse to produce antiserum.

[0040] Cell fusion and cloning: Splenocytes from immunized Balb / c mice were fused with SP2 / 0 myeloma cells at a ratio of 9:1 to obtain hybridoma cell lines of monoclonal antibodies.

[0041] Cell cryopreservation and recovery: the hybridoma cells were made into 1×10 cryopreservation medium 9 cells / ml for long-term storage in liquid nitrogen. When recovering, take out the cryopreservation tube, put it into a 37°C water bath to thaw quickly, remove the cryopreservation solution by centrifugation, and transfer it to a culture bottle for culture.

[0042] Preparation and...

Embodiment 2

[0052] The formation of embodiment two kits

[0053] The magnetic particle chemiluminescence kit for detecting chlorpromazine was assembled so that it contained the following components:

[0054] Fluorescent marker of FITC-labeled chlorpromazine monoclonal antibody

[0055] Luminescent marker of ABEI-labeled chlorpromazine hapten

[0056] Separation reagent of paramagnetic nanobeads coated with goat anti-FITC monoclonal antibody

[0057] Chlorpromazine standard solution (0ng / ml, 0.01ng / ml, 0.03ng / ml, 0.09ng / ml, 0.27ng / ml, 0.81ng / ml), the standard diluent is pH7.4, 0.03%NaN 3 , 0.05mol / L TRIS buffer. The percentage content is a mass percentage content.

[0058] The concentrations of the chlorpromazine quality control solution are 0.02ng / ml and 0.5ng / ml respectively, and the quality control dilution solution is pH7.4, 0.03%NaN 3 , 0.05mol / L TRIS buffer. The percentage content is a mass percentage content.

[0059] Concentrated lotion pH7.2, 0.2-0.4% Tween-20, 0.02-0.04% N...

Embodiment 3

[0060] The detection of chlorpromazine in embodiment three actual samples

[0061] 1. Sample pretreatment

[0062] (1) Pretreatment method of chicken and pork samples

[0063] Weigh 1.0±0.05g homogeneous material into a 50ml polystyrene centrifuge tube, add 3.2ml acetone, then add 0.8ml 0.2M hydrochloric acid solution, shake vigorously with an oscillator for 5min; over 3000g, centrifuge at room temperature (20-25°C) 10min; pipette 2ml supernatant and add 4ml 2M sodium hydroxide solution to mix, let it stand at room temperature for 10min, then add 6.4ml n-hexane, shake with an oscillator for 5min, centrifuge at room temperature (20-25℃) for 5min at 3000g or more; take 4ml Put the upper layer into a 10ml clean glass test tube, and dry it under nitrogen flow in a water bath at 50-60°C; add 1ml of reconstitution working solution and vortex for 2 minutes to dissolve the dried residue; take 50μl for analysis, sample dilution factor: 4.

[0064] (2) Feed sample pretreatment method ...

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Abstract

The present invention relates to a magnetic particle chemiluminescence kit for detecting chlorpromazine. The kit comprises a luminous marker, a fluorescein marker, a standard substance, a quality control substance and a separation reagent, wherein the luminous marker is isoluminol luminous marker labeled chlorpromazine hapten, the fluorescein marker is fluorescein labeled chlorpromazine monoclonal antibody or fluorescein derivative labeled chlorpromazine monoclonal antibody, and the separation reagent is sheep anti-FITC monoclonal antibody coated paramagnetic nanometer microbeads. The present invention further relates to a method for detecting chlorpromazine in foods of animal origin by using the kit, wherein the method provides characteristics of high sensitivity, high specificity and rapid detection for chlorpromazine detection.

Description

technical field [0001] The invention relates to a direct chemiluminescence detection kit and a detection method thereof. Especially the magnetic particle competition direct chemiluminescent detection kit for the detection of chlorpromazine residues in animal tissue, liver, feed and other samples. technical background [0002] Chlorpromazine is a representative drug of phenothiazines, a blocker of central dopamine receptors, and has various pharmacological activities such as antipsychotic, antiemetic, cooling and enhancing hypnotic anesthesia. Use during animal feed transportation can reduce animal maintenance needs and reduce weight loss and mortality on the way. In recent years, some feed companies have pursued the conversion rate and high profits of feed, and randomly added sedative and hypnotic drugs to feed, and the residues in animal tissues lead to unsafe products. [0003] At present, the main methods for detecting chlorpromazine include high performance liquid chro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577
Inventor 万宇平罗晓琴韩锐付军权杨学林刘琳
Owner BEIJING KWINBON BIOTECH
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