Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof

An artificial antigen, chlorpromazine technology, applied in chemical instruments and methods, animal/human protein, serum albumin, etc., can solve the problems of lack of chlorpromazine, chlorpromazine antibody specificity and insufficient recognition, and achieve The effect of high accuracy, high sensitivity and high specificity

Active Publication Date: 2012-11-07
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Immunological analysis technology has been widely used in the field of drug residue detection due to its advantages of sensitivity, specificity, rapidity and simplicity. The synthesis of artificial antigen is the key to the establishment of hapten substance immunoassay method. For chlorpromazine, because the hapten design and synthetic reasons, the specificity and recognition of the chlorpromazine antibody obtained by the prior art are not enough, and the relevant technical reports of the immunological analysis method of chlorpromazine are lacking

Method used

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  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof
  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof
  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Hapten Synthesis and Identification

[0034] 1. Synthesis of chlorpromazine hapten

[0035] The synthetic route of chlorpromazine hapten is attached figure 1 shown.

[0036] (1) Take by weighing 319mg (1mmol) chlorpromazine and dissolve in 5mL acetic anhydride, stir and dissolve fully in an ice bath;

[0037] (2) Take 2 mmol of freshly prepared acetyl nitrate (refer to the routine for the preparation method) and dissolve it in 1 mL of acetic anhydride and slowly drop it into the solution described in step (1), and react in an ice bath for 1 hour. Sodium oxide solution adjusted the pH value to 8, crystallized to obtain 2-chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxo-10H-phenothiazine, the yield was 61 %;

[0038] (3) Take the above-mentioned 2-chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxo-10H-phenothiazine and dissolve it in 20mL of ethanol, add 2 -Chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxygen-10H-phenothiazine 10% (according to mass percen...

Embodiment 2

[0042] Example 2: Synthesis and identification of artificial antigens

[0043] 1. Synthesis and identification of chlorpromazine antigen (III)

[0044] Chlorpromazine antigen (III) glutaraldehyde synthesis:

[0045] (1) Take by weighing 35mg chlorpromazine hapten and 68mg BSA and be dissolved in 10mL phosphate buffer saline (PBS) solution together, stir at room temperature;

[0046] (2)) Add 40 μL of glutaraldehyde dropwise into the mixed solution described in step (1), and stir and react at room temperature for 3 hours after the dropwise addition;

[0047] (3) Put the solution reacted in step (2) into a dialysis bag, dialyze with PBS solution for 72 hours at 4°C, change the dialysate 6 times during the period, and obtain the target product, which is stored at -20°C after aliquoting.

[0048] In the above method, human serum protein, hemocyanin or ovalbumin can also be used. Referring to the method of this embodiment, the preparation method of chlorpromazine-coated antigen is ...

Embodiment 5

[0051] Preparation and Identification of Embodiment 5 Chlorpromazine Antigen

[0052] 2. Synthesis and identification of chlorpromazine antigen (II)

[0053] The diazotization method synthesis of chlorpromazine antigen (II):

[0054] (1) The chlorpromazine hapten that takes 35mg is dissolved in the hydrochloric acid of 1mL 1mol / L, and the hydrochloric acid adjustment solution pH value is 2 with the hydrochloric acid of 1mol / L;

[0055] (2) Take 1.1 mL of 1mol / L sodium nitrite solution and add it dropwise to the hapten solution within half an hour, and diazotize at 0-5°C for 1 hour;

[0056] (3) Weigh 68mgBSA and dissolve it in 10mL of PBS (phosphate buffer saline used routinely in the laboratory) solution, add the solution after the reaction in step (2) dropwise into the BSA-PBS solution, and use 1mol / L sodium hydroxide Adjust the pH value of the solution to 7.5, overnight at 4°C;

[0057] (4) Put the solution reacted in step (3) into a dialysis bag, dialyze with PBS soluti...

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PUM

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Abstract

The invention provides a chlorpromazine hapten, an artificial antigen and an antibody as well as a preparation method and application thereof. The chlorpromazine artificial hapten has a molecular structure (I); the chlorpromazine artificial antigen has molecular structures (II) or (III); the antiserum obtained by the invention has the titer of 1:3*105, the lowest detection limit of 3ng / mL and thehalf-inhibition concentration of 29ng / mL; and the generated antibody has high specificity, sensitivity and accuracy. The antigen and the antibody have wide application prospect in the detection of residual chlorpromazine in the food.

Description

technical field [0001] The invention belongs to the technical field of food safety. It specifically relates to the preparation method and application of chlorpromazine hapten, artificial antigen and antibody. Background technique [0002] Chlorpromazine (Chlorpromazine, CPZ) is also known as Dongmianling, and its molecular formula is: C 17 h 19 C 1 N 2 S, generally in the form of hydrochloride for medicinal use, the chemical name is: N, N-dimethyl-2-chloro-10H-phenothiazine-10-propylamine hydrochloride. Belonging to one of the phenothiazine drugs, it is a central dopamine receptor blocker with sedative, antipsychotic, antiemetic, hypothermic and basal metabolism, -Adrenergic receptor and m-cholinergic receptor blockade, antihistamine, endocrine effects, etc. It is clinically used to control symptoms such as restlessness, nervousness, hallucinations, and delusions of schizophrenia or other mental illnesses; to treat vomiting caused by various reasons; it is also used f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D279/34C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/566G01N33/577
Inventor 孙远明沈玉栋徐振林王弘肖犇雷红涛杨金易
Owner SOUTH CHINA AGRI UNIV
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