Time-resolved immunoassay kit for detecting chlorpromazine residues and detection method thereof

A detection kit and immunoassay technology, applied in the field of immunoassay, can solve the problems of lack of chlorpromazine time-resolved immunoassay kit, etc., and achieve the effect of simple sample pretreatment, short operation process and good effect

Inactive Publication Date: 2012-07-04
FOSHAN ZHONGNAN AGRI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, there is currently no relevant technical report on the detection of chlorpromazine by time-resolved immunoassay, and there is no kit suitable for the detection of chlorpromazine by time-resolved immunoassay, so as to realize high-sensitivity, simple-to-operate large-scale rapid detection

Method used

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  • Time-resolved immunoassay kit for detecting chlorpromazine residues and detection method thereof
  • Time-resolved immunoassay kit for detecting chlorpromazine residues and detection method thereof
  • Time-resolved immunoassay kit for detecting chlorpromazine residues and detection method thereof

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Embodiment 1

[0044] A time-resolved immunoassay detection kit for detecting chlorpromazine residues, comprising a microtiter plate coated with chlorpromazine antigen, chlorpromazine antibody, lanthanide-labeled goat anti-rabbit or goat anti-mouse antibody (secondary antibody) ; the lanthanides include Eu 3+ , Tb 3+ 、Dy 3+ 、Sm 3+ and other rare earth elements.

[0045] The chlorpromazine antigen is obtained by coupling the chlorpromazine hapten with the carrier protein by carbodiimide method (EDC), active ester method (DCC, NHS), mixed anhydride method (isobutyl chloroformate). The carrier protein includes bovine serum albumin (BSA), human serum albumin (HSA), keyhole limpet hemocyanin (KLH) and other carrier proteins.

[0046] The invention provides a preparation method of the chlorpromazine hapten, which is to dissolve chlorpromazine in acetic anhydride, drop freshly prepared acetyl nitrate in an ice bath, and react to obtain CPZ-NO 2 , and then hydrogen reduction under the catalysis...

Embodiment 2

[0080] The present invention provides the method for detecting chlorpromazine residue simultaneously, comprises the following steps:

[0081] (1) Sample pretreatment;

[0082] a. Urine sample processing

[0083] Clear urine samples can be directly analyzed. If the urine sample is turbid, centrifuge at 2000r / min for 5min or filter, and use the supernatant for detection.

[0084] b. Feed sample processing

[0085] Crush the feed, weigh 2 g and place it in a 50 mL test tube, add 12 mL of 10% ammoniated methanol (9 parts of ammonia solution with a pH value of 9 to 10 + 1 part of methanol), mix well and shake for 1 to 2 minutes. Add 9 mL of ethyl acetate and shake for 20 min. Centrifuge at 5000r / min (4000g) for 10min. Take 500 μL of the organic phase in another test tube and dry it with nitrogen. Add 500 μL of sample diluent to dilute and detect directly. A dilution factor of 10 should be taken into account in the calculation of results.

[0086] c. Tissue (muscle, liver, k...

example 1

[0105] Hapten synthesis and identification

[0106] The synthesis of chlorpromazine hapten as figure 2 shown.

[0107] (1) Take by weighing 319mg (1mmol) chlorpromazine and dissolve in 5mL acetic anhydride, stir and dissolve fully in an ice bath;

[0108] (2) Get freshly prepared acetyl nitrate 4mmol and dissolve it in 1 mL of acetic anhydride and slowly drop it into the solution described in step (1), react in an ice bath for 3 hours, and after the reaction is completed, adjust the pH value of the product with ice-saturated sodium hydroxide solution To 8, the precipitated product is the nitrated compound of chlorpromazine, and the productive rate is 53%;

[0109] (3) Dissolve the nitrate of the above-mentioned chlorpromazine in 20mL ethanol, add 10% palladium carbon catalyst and obtain the target product under hydrogen reduction.

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Abstract

The invention provides a time-resolved immunoassay kit for detecting chlorpromazine residues and a detection method thereof. The kit of the invention comprises an ELISA (enzyme-linked immunosorbent assay) coated with chlorpromazine antigen, lanthanide element-labeled goat anti-rabbit or goat anti-mouse antibodies, anti-chlorpromazine antibodies, and the like. The invention also discloses a method for detecting chlorpromazine residues using the above kit. The kit for detecting chlorpromazine residues provided by the invention adopts indirect competitive time-resolved immunoassay technology, has high sensitivity and good stability, greatly simplifies operation procedures and reaction time, reduces errors due to complex operation, reduces cost, is suitable for screening of a large number of samples, and has very important operation significance.

Description

technical field [0001] The invention belongs to the technical field of immunodetection, and in particular relates to a time-resolved immunoassay kit for detecting residual chlorpromazine and a preparation method thereof. Background technique [0002] In recent years, with the rapid development of social economy and the intensive development of breeding industry, the application of prohibited additives in animal husbandry production has increased significantly, and the resulting problem of drug residues in animal foods has become increasingly prominent, such as "lean Accidents such as "Meat Essence Incident", "Melamine Incident" and "Shuanghui Incident" occurred frequently. Drug residues in animal foods are a major hidden danger to the health of consumers, and severe cases can cause poisoning or even death. These incidents not only caused great losses to the export of animal-derived food in our country, but also seriously affected the health of the people. [0003] Chlorpro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/544G01N1/28G01N1/38
Inventor 伍志权朱晓明邢贞妙彭子强李红苑黄换开
Owner FOSHAN ZHONGNAN AGRI & TECH
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