A kind of preparation method of caffeine artificial antigen

A technology of artificial antigen and artificial hapten, which is applied in the field of preparation of caffeine artificial antigen, can solve the problems of expensive equipment, time-consuming detection, and unreachability, and achieve the effect of advanced synthesis technology and strong specificity

Active Publication Date: 2018-02-13
HANGZHOU ALLTEST BIOTECH
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  • Application Information

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Problems solved by technology

[0005] At present, the detection of caffeine mainly relies on high-performance liquid chromatography (HPLC), gas chromatography (GC), thin-layer chromatography (TLC), mass spectrometry (MS), etc., but there are expensive instruments, time-consuming detection, and professional and technical personnel operation, can not meet the requirements of modern detection for fast and accurate

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  • A kind of preparation method of caffeine artificial antigen
  • A kind of preparation method of caffeine artificial antigen
  • A kind of preparation method of caffeine artificial antigen

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Embodiment 1

[0032] (1) Preparation of artificial hapten:

[0033](a) Add 300mg (1.67mmol) of 3,7-dimethylxanthine and 333mg (3.33mmol) of succinic anhydride into a round bottom flask, dissolve them with pyridine, and react under reflux at 100°C for 16 hours; Cooled to room temperature, dried, and separated by thin-layer chromatography to obtain 213 mg of white solid product I; thin-layer chromatography: the chromatographic solution was 95% ethanol: 1,4-dioxane: dichloromethane: ammonia water = 8:1: 10:1, product R f =0.4;

[0034] (b) Add 117mg (0.76mmol) of 3-aminodihydrothiophene-2-one hydrochloride and 213mg (0.76mmol) of product I into a round bottom flask, add 0.2ml pyridine, 4.56ml N,N-dimethylformaldehyde Amide (DMF) was dissolved, then added 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride 168mg (0.87mmol), N 2 Under the atmosphere, the reaction was stirred at room temperature for 1 hour; after the reaction, the solvent was dried. Add 16ml of water to the obtained re...

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Abstract

The invention provides a preparation method of a caffeine artificial antigen. The preparation method comprises the following steps: step 1, preparation and detection of hapten: using 3,7-dimethylxanthine and butanedioic anhydride as raw materials, performing an acylation reaction to obtain an intermediate containing carboxyl, performing the acylation reaction to the intermediate and 3-amino-dihydrothiophene-2-one, and performing hydrolysis to obtain the hapten containing sulfydryl; step 2, preparation and detection of artificial antigen: firstly using an N-hydroxyl succinimide active ester method to activate bromoacetic acid, then performing crosslinking with bovine serum albumin (BSA) to obtain the bovine serum albumin containing bromo chain hydrocarbon, performing a condensation reaction with the bovine serum albumin and the sulfydryl containing hepten obtained in the first step to prepare the caffeine artificial antigen, namely caffeine-bovine serum albumin. The prepared caffeine artificial antigen can be used for animal immunization to obtain corresponding caffeine antigens, and further can be used for study of various caffeine immunoassays, so as to provide an approach which is more convenient, faster and more accurate for caffeine detection.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a preparation method of caffeine artificial antigen. Background technique [0002] Caffeine (caffeine) name 1,3,7-trimethylxanthine, also known as caffeine, tea, mateine, guarana factor. It is a xanthine alkaloid compound that can be used as an insecticide. For humans, moderate consumption can relieve fatigue and excite nerves. It is clinically used to treat neurasthenia and coma recovery. Its structural formula is: [0003] [0004] Caffeine is a central nervous system stimulant, which is addictive. Large doses or long-term use can also cause damage to the human body, especially it is also addictive. Withdrawal symptoms, when the dosage of the drug continues to increase, caffeine will not only act on the cerebral cortex, but also directly excite the medulla oblongata, causing paroxysmal convulsions and bone tremors, damaging important internal organ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765
CPCC07K14/765
Inventor 王镇高飞邵越水
Owner HANGZHOU ALLTEST BIOTECH
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