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F-Ketamine hapten, F-Ketamine antigen as well as preparation method and application thereof

A technology of fluoroamine and hapten, which is applied in the biological field, can solve the problems that cannot meet the requirements of rapidity, convenience, and accuracy, and cannot meet the detection standard of fluoroamine, and achieves the reduction of cross-substance interference, mild process conditions, and easy operation. simple effect

Pending Publication Date: 2021-01-05
GUANGZHOU WONDFO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At this stage, China has not been included in the national control drug list, and there is no clear regulation and description for this compound. However, there are a large number of abusers who use fluoroamine instead of K powder in the society. The detection of fluoroamine is limited to high-efficiency 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 Expensive instruments and equipment are required, and the requirements for inspection materials are relatively high, and further purification is required to carry out, which can no longer meet the requirements of modern detection for fast, convenient, and accurate
[0004] However, the use of ketamine rapid detection reagents does not meet the standards for the detection of fluoroamine

Method used

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  • F-Ketamine hapten, F-Ketamine antigen as well as preparation method and application thereof
  • F-Ketamine hapten, F-Ketamine antigen as well as preparation method and application thereof
  • F-Ketamine hapten, F-Ketamine antigen as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] This embodiment prepares a kind of fluoroamine hapten, and the preparation method is as follows:

[0066] Weigh 10mmol carboxymethyl hydroxylamine hydrochloride and 10mmol K 2 CO 3 In a 50mL round-bottomed flask, add 21mL methanol and 9mL purified water, stir well for 20min, add 0.1mmol 18-crown-6 and 10mmol fluoroamine, and reflux at 65°C for 24h; after the reaction, evaporate under reduced pressure Dry, add 50mL ultrapure water and 50mL dichloromethane to the residue, separate the water phase and adjust the pH to 4 with 2M hydrochloric acid, extract twice with 10mL n-butanol, combine the n-butanol layers and carry out dry and vacuum distillation to obtain 2.3 The mmol hapten is ready for use; the prepared hapten is analyzed by ESI-MS (295.1 [M+1]), the result is as follows figure 1 As shown, it was proved that the hapten of fluoroamine was successfully prepared.

Embodiment 2

[0068] In this example, a fluoroamine antigen is prepared, and the preparation method is as follows:

[0069] (1) Preparation of Fluoramine hapten active ester:

[0070] In a 25mL dry single-necked bottle, add 1mmol of the hapten prepared in Example 1, 1.2mmol of N-hydroxysuccinimide, 1.2mmol of cyclohexylcarbodiimide and 8mL of N,N-dimethylformamide. Stir at 25°C for 18h under the protection of helium. After the reaction was completed, the reaction mixture was divided into eight 1.5mL centrifuge tubes, centrifuged at 10000rpm for 30min, and the supernatant was divided into eight 2mL glass sealed bottles and stored under the protection of helium gas for later use.

[0071] (2) Preparation of fluoroamine antigen (fluoroamine-bovine serum albumin):

[0072] Take 100mg of BSA in a 25mL one-mouth bottle, add 8mL of 0.1M phosphate buffer with pH 7.0 and stir to dissolve, use an ice-water bath to keep the temperature of the reaction mixture at about 4°C, and slowly add 2.6mL of ac...

Embodiment 3

[0079] This embodiment prepares an immunochromatographic diagnostic reagent strip and reagent card for detecting fluoroamine in urine samples, and the preparation method is as follows:

[0080] (1) Sample pad preparation: Soak the glass fiber in the sample pad treatment solution, and then dry it. The sample pad treatment solution is a formula of 0.02M pH 8.4 Tris and contains the following components by mass fraction: 0.8% Triton X-100, 0.8 %Tween-80, 2% rabbit serum, 1% HPMC, 1% trehalose, 1% LowCross-Buffer, 2% NaCl, 1% Na 2 CO3 , 0.5‰Proclin 300.

[0081] (2) Marking pad preparation: take the mass fraction of 0.01% HAuCl 4 100mL of aqueous solution, add 0.75mL of 1% trisodium citrate aqueous solution by mass fraction, heat and boil for 30min, the color turns red. Turn off the heat and let cool for later use. Colloidal gold solution with 0.1MK 2 CO 3 Adjust the pH to 9.0, stir the colloidal gold solution, add anti-flutamine monoclonal antibody (provided by Guangzhou Wan...

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Abstract

The invention relates to a F-Ketamine hapten, a F-Ketamine antigen as well as a preparation method and application thereof. The structure of the F-Ketamine hapten is shown as a formula (I). The preparation method of the F-Ketamine hapten comprises the following steps: taking F-Ketamine and a compound shown as a formula (II) as raw materials, carrying out reacting, and introducing an active arm with carboxyl on carbonyl of the F-Ketamine to obtain the F-Ketamine hapten. Furthermore, after the F-Ketamine hapten is activated, the F-Ketamine hapten and carrier protein are subjected to a coupling reaction to obtain the F-Ketamine antigen. The invention initiates a novel F-Ketamine artificial hapten and a novel F-Ketamine artificial antigen for the first time; and the novel F-Ketamine artificialhapten and the novel F-Ketamine artificial antigen can be applied to an immunochromatography technology, have the advantages of high sensitivity, high specificity, simplicity, rapidness, easiness inoperation, no need of any large-scale instruments and equipment and the like, and can be used for large-scale preliminary screening of F-Ketamine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a fluoroamine hapten, a fluoroamine antigen, a preparation method and an application thereof. Background technique [0002] Fluoroamine (F-Ketamine) is an amphetamine-type stimulant, and its chemical name is: dl-2-o-fluorophenyl-2-methylaminocyclohexanone. Fluoramine, a derivative of phencyclidine, has a certain psychotropic dependence potential. Fluoramine can produce a state of dissociative anesthesia, and its clinical features are similar to those of ketamine (K powder), including stiffness, light sedation, amnesia and significant analgesia, and can enter dreams and hallucinations, becoming a wild party and other similar symptoms. Psychedelic substances commonly used in activities. At present, many countries have listed fluoroquinone, the United Kingdom has listed fluoroamine in CLASS B, and Sweden and Latvia have defined fluoroquinone as an illegal ingredient. Professor Wang Shi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C251/60C07C249/08C07K14/47C07K14/765C07K14/77C07K14/795C07K14/805C07K16/00C07K1/34C07K1/107G01N33/558G01N33/58G01N33/94
CPCC07C251/60C07C249/08C07K14/765C07K14/77C07K14/795C07K14/805C07K14/47C07K16/00G01N33/946G01N33/558G01N33/585C07C2601/14
Inventor 袁强王平梁飞敏
Owner GUANGZHOU WONDFO BIOTECH
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