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Preparation method for ethyl urethane detection test paper and detection method of test paper

A technology of urethane and test paper, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems that the accuracy cannot be compared with that of GC-MS, there are many impurities such as pigments, and cumbersome pretreatment steps, etc., and the detection method is simple. Rapid, simplified detection procedures, and small volume of test paper

Inactive Publication Date: 2012-10-17
ZHEJIANG FORESTRY UNIVERSITY
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Problems solved by technology

[0011] (1) Dichloromethane extraction - GC-MS technology: After diluting the test sample with known alcohol concentration with water, wash with n-hexane, then extract, concentrate, and constant volume with dichloromethane, and then use gas chromatography-mass spectrometry Detector determination, external standard method for quantification, the detection limit is 10 μg kg -1 , the recovery rate of this method is low, and there are more impurities such as pigments
[0012] (2) Solid Phase Extraction-Gas Chromatography Technology: Using solid phase extraction technology to extract and purify ethyl carbamate in alcoholic beverages, and then using GC / MS for qualitative and quantitative analysis, the accuracy and efficiency of the method have been greatly improved. It has been greatly improved, but the extraction column needs to be cleaned every time it is used, which brings certain difficulties to the determination
Studies have shown that the accuracy of this method cannot be compared with that of GC
[0015] The common disadvantages of the above-mentioned qualitative or quantitative detection of ethyl carbamate are that it involves a large number of instruments and cumbersome pretreatment steps, the detection cycle is too long, and it is easy to introduce analysis and measurement errors, which is not suitable for rapid detection.

Method used

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  • Preparation method for ethyl urethane detection test paper and detection method of test paper

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Embodiment Construction

[0029] Below in conjunction with implementation example the present invention is described in further detail:

[0030] The preparation method of this urethane test paper is carried out as follows:

[0031] (1) Material preparation: acetylcholinesterase with activity ≥ 220u / g, ethyl carbamate with a purity of 99%, the paper base of the test paper is a medium-speed qualitative filter paper with good water absorption, and the substrate is 1g L -1 2,6-dichloroindophenol acetate;

[0032] (2) Preparation of enzyme solution: 10 u of acetylcholinesterase, 2 μL of glutaraldehyde with a volume ratio of 5%, and 10 μL of BSA with a mass ratio of 1%, were prepared into a 70 μL enzyme solution;

[0033] (3) Preparation of test paper: the above-mentioned enzyme solution and qualitative filter paper were routinely immobilized at 3° C. for 8 hours to obtain urethane detection test paper.

[0034] The method that this urethane test paper detects is carried out as follows:

[0035] (1) Prepa...

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Abstract

The invention provides a preparation method for ethyl urethane detection test paper. The preparation method comprises three steps as follows: material preparation: purchasing acetylcholinesterase with activity no less than 220u / g, ethyl urethane with purity of 99%, medium-speed qualitative filter paper with good hygroscopicity, and 2-6-dichlorophenolindophenol acetate; preparing esterase solution; and preparing immobilization of the test paper. The detection method by using the test paper prepared by the method comprises four steps as follows: preparing standard detection solution; preparing a standard colourimetric card; confirming the effectiveness of the test paper; and framing the colorimetry and content. The test paper has the advantages of being fast and convenient, and capable of judging only through visual observation when being used for detecting and removing ethyl urethane harmful substances in alcohol drinks, fermented food and the like, thus not only being applicable in food detection departments, as well as in food production enterprises, sales departments, and users capable of conducting self-detection, and application and popularization range is wide.

Description

technical field [0001] The invention relates to a preparation and detection method of a quick qualitative and semi-quantitative detection test paper for ethyl carbamate in food, especially fermented food. Background technique [0002] The molecular formula of urethane (EC) is C 3 h 7 NO 2 , not only a natural component of tobacco, but also present in fermented foods (yogurt, soy sauce, etc.) and alcoholic beverages (rice wine, sake, wine, etc.). The main way for the human body to absorb EC is to drink alcoholic beverages. Studies have shown that about 0.5% of ethyl carbamate is oxidized to acetyl carbamate by cytochrome P450 in the organism, and then further forms acetyl carbamate epoxide, and the epoxide formed by this way can be in The formation of DNA addition polymers in organisms leads to damage to the double strands of the DNA structure, which in turn leads to canceration of cells; about 0.1% of urethane is oxidized to N-hydroxyl urethane by cytochrome P450, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52
Inventor 郭明严锋李博斌敬娇梁东军
Owner ZHEJIANG FORESTRY UNIVERSITY
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