Method for quickly measuring ethyl carbamate in liquor samples in batches

A kind of urethane, rapid determination technology, applied in the field of analytical chemistry and food safety detection, can solve the problem of inconsistent pH in the optimal reaction of alanine dehydrogenase, and achieve the effect of low cost

Active Publication Date: 2017-11-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to solve the problem of inconsistency in the optimum reaction pH between the EC degrading enzyme and the modified alanine dehydrogenase on the enzyme electrode, a two-stage enzyme coupling reaction method was designed.

Method used

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  • Method for quickly measuring ethyl carbamate in liquor samples in batches
  • Method for quickly measuring ethyl carbamate in liquor samples in batches
  • Method for quickly measuring ethyl carbamate in liquor samples in batches

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Determination of the optimal concentration of chitosan (CS) and reduced graphene oxide (RGO) in the preparation of screen-printed electrodes modified by alanine dehydrogenase (AlaDH)

[0021] When preparing alanine dehydrogenase-modified SPE, it is necessary to optimize the concentration of CS and RGO for immobilizing AlaDH to the electrode surface. The concentration of CS was optimized using the method of single factor experiment. First, prepare CS with mass concentrations of 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, and 0.6%, add an appropriate amount of glycerol as a protective agent, stir evenly to remove foam, and place in a 4°C refrigerator for later use. Take 50 μL of CS solutions of different concentrations, mix them with 25 μL of 20 U / mL AlaDH respectively, and take 20 μL of the mixed solution and apply it dropwise on the surface of SPE. The prepared electrode was used to detect the standard EC solution according to the above two-stage reaction, and the current ...

Embodiment 2

[0022] Embodiment 2. Determination of the optimum conditions for the first-stage reaction

[0023]The enzyme activity of EC degrading enzyme added to the reaction system was optimized by single factor test. Add 6, 8, 10, 12, 14, 16 U / mL of EC degrading enzymes to the solution containing 8mmol / L EC respectively. After reacting for 1 h, adjust the pH to 10 and add 50 mmol / L sodium pyruvate solution and 0.4 The mmol / L NADH solution was measured with a modified electrode, and the current response change value of NADH was recorded within 5 minutes of reaction time. When the addition amount of EC degrading enzyme was 16 U / mL, the change value of the electrode current response was the largest, and taking into account the use cost of the enzyme, the optimal addition amount of EC degrading enzyme was determined to be 16 U / mL.

[0024] The reaction time of the first stage was optimized by single factor experiment. Add 16 U / mL of EC degrading enzyme to the solution containing 8 mmol / L ...

Embodiment 3

[0025] Embodiment 3. Determination of the optimum conditions for the second stage reaction

[0026] After using the optimal conditions for the first-stage reaction, the concentration of NADH in the second-stage reaction system was optimized by single factor test. After the solution after the first stage reaction was adjusted to pH 10, NADH and 50 mmol / L sodium pyruvate were added at a concentration of 0.1, 0.2, 0.3, 0.4, 0.5, 0.75, 1.0 mmol / L, and the prepared alanine was used to remove The hydrogenase-modified screen-printed electrode was used for amperometric time measurement, and the optimal concentration of NADH was determined to be 0.5 mmol / L according to the size of the NADH response change value.

[0027] The concentration of sodium pyruvate added to the reaction system in the second stage was optimized by single factor experiment. After adjusting the pH of the solution after the first stage reaction to 10, add sodium pyruvate solution and 0.5 mmol / L NADH solution with...

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Abstract

The invention discloses a method for quickly measuring ethyl carbamate in liquor samples in batches, and belongs to the technical field of analytical chemistry and food safety detection. The method comprises the following steps: firstly, pretreating the liquor samples with a EC (ethyl carbamate) special solid phase column to extract EC in the liquor samples; then adding EC degrading enzyme into a solution system obtained by extraction; finally, adding sodium pyruvate and NADH which serve as matrixes into a detection system, adjusting the pH with a buffer solution, performing electrochemical detection by utilizing an AlaDH-modified silk-screen printing electrode, and measuring a linear relation between a current response value drop caused by consumption of the NADH and the concentration of the EC with a current-time method, thus measuring the EC content. According to the method, an EC sensor based on the enzyme-modified silk-screen printing electrode is built and is linked with the special solid phase column for EC extraction, so that influence of urea which is a main interfering substance in a liquor matrix is effectively eliminated, and direct measurement of the EC content of each liquor sample such as yellow wine is realized. Compared with a GC-MS (Gas Chromatography-Mass Spectrometer) serving as a principal method for EC measurement at the present, the method disclosed by the invention has the advantages that no large-sized instrument equipment is needed, and fast, high-sensitivity and low-cost batch measurement can be realized.

Description

technical field [0001] The invention relates to a batch method for rapid determination of ethyl carbamate in wine samples, which can perform high-sensitivity and rapid determination of ethyl carbamate in wine samples such as rice wine in batches, and belongs to analytical chemistry and food safety detection technology field. Background technique [0002] Ethyl carbamate (EC), commonly known as urethane and urethane, is a colorless and odorless crystal. It is a harmful metabolite produced during the production of fermented food and beverages. It not only exists in fermented wine, It also exists in fermented foods such as bread and soy sauce. EC is a multi-site carcinogen that can cause lung cancer, liver cancer and other diseases in rodents. Now it is generally believed that the main mechanism of EC carcinogenesis is that when copper ions exist, EC interacts with cytochrome P450 in the body, causing DNA damage, thereby causing liver, Cancer of lungs and other organs. Based...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N1/34G01N1/40C12Q1/00
CPCC12Q1/005G01N1/34G01N1/405G01N27/26G01N27/30G01N2333/90616
Inventor 周楠迪张智威蔡蓉凤田亚平
Owner JIANGNAN UNIV
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