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Synthetic method of urethane artificial antigen

A urethane, artificial antigen technology, applied in chemical instruments and methods, animal/human proteins, serum albumin, etc., can solve the problem of undetected coupling ratio, raw material dosage based on experience, difficult urethane Fast and economical detection and other problems, to achieve the effect of small shielding effect and accurate dosage

Inactive Publication Date: 2012-10-10
ZHEJIANG FORESTRY UNIVERSITY
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Problems solved by technology

[0003] At present, the determination method of ethyl carbamate is mainly based on gas chromatography analysis, coupled with various detection methods, including: gas chromatography-hydrogen flame ionization detector (GC-FID), gas chromatography / thermal ion detector method (GC-TSD), dichloromethane extraction and gas chromatography-mass spectrometry (GC-MS), etc., such as 200910099554.0 "a method for detecting ethyl carbamate in rice wine", 201110339615.3 "a rapid determination of distilled wine The methods for the content of ethyl carbamate in medium and medium-sized urethanes” all adopt the pretreatment process and large-scale instrument determination, and it is difficult to realize the rapid and economical detection of urethane
In 200710190350.9 "Highly sensitive time-resolved fluorescent immunoassay method for ethyl carbamate in alcohol", although the follow-up immunological operation was carried out, the coupling ratio was not detected during the artificial antigen synthesis process, and the amount of raw materials was based on experience. theoretical support

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  • Synthetic method of urethane artificial antigen
  • Synthetic method of urethane artificial antigen
  • Synthetic method of urethane artificial antigen

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Embodiment Construction

[0025] The present invention is described in further detail below in conjunction with embodiment and with reference to accompanying drawing:

[0026] Specific embodiments are described below.

[0027] Preparation:

[0028] Bovine serum albumin and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide are products of Sigma; ethyl carbamate is a product of Tsi Ai Chemical Industry Development Co., Ltd.; other reagents used in the experiment are analytical pure.

[0029] Instruments selected:

[0030] IR Prestige-21 Fourier Transform Infrared Spectrometer is a product of Shimadzu, Japan; AVANCE AV-300 NMR Spectrometer is a product of Swiss BRUKER; Voyager-DE STR matrix-assisted laser desorption time-of-flight mass spectrometry is a product of Applied Biosystems, Inc.

[0031] 1. Hapten Preparation

[0032] (1) Weigh 4.45g β-alanine, dissolve in 100mL NaHCO with a mass volume ratio of 4% 3 In aqueous solution, as reagent A, 4.14 g of ethyl chloroformate was dissolved in 30 mL of chlo...

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Abstract

The invention provides a synthetic method of urethane artificial antigen. The urethane artificial antigen is prepared by the following steps of: 1, preparation of beta-alanine and ethyl chloroformate; 2, synthesis of hapten; 3, purification of hapten; 4, preparation of 1-(3-dimethylamino propyl)-3-ethyl carbodiimide solution; 5, synthesis of artificial antigen; 6, dialysis and detection of artificial antigen; and 7, calculation of quantity of different used carrier proteins. According to the synthetic method, the shielding effect of the protein to the hapten obtained by chemical synthesis is low, therefore, high-titer polyclonal antibody can be generated in an immune animal, and the molecular weight can be detected during synthesizing the artificial antigen, and the coupling ratio can be calculated, and the quantity of used raw materials for synthesizing is precise.

Description

technical field [0001] The invention relates to a method for synthesizing an artificial antigen, in particular to a method for synthesizing an ethyl carbamate artificial antigen. Background technique [0002] Ethyl Carbamate (EC), also known as urethane and urethane, is a naturally occurring pollutant in food and alcoholic beverages during fermentation or storage. As early as the 1940s, urethane was confirmed as a carcinogen by Nettleship. In 2007, the International Agency for Research on Cancer reclassified urethane as Group 2A. Studies have shown that urethane, as a multi-site carcinogen, can cause diseases such as lung cancer, lymphoma, liver cancer and skin cancer in rodents, and it has also been found that ethanol can promote the carcinogenicity of urethane. [0003] At present, the determination method of ethyl carbamate is mainly based on gas chromatography analysis, coupled with various detection methods, including: gas chromatography-flame ionization detector (GC-F...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/77C07K14/795
Inventor 郭明周伟敬娇谢广发周建钟
Owner ZHEJIANG FORESTRY UNIVERSITY
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