Study On Rapid Detection Of Ethyl Carbamate In Alcoholic Beverages Based On Enzyme Inhibition Method

In this paper, the quantitative analysis method of spectrophotometry was used to detectethyl carbamate in alcoholic beverages based on enzyme inhibition, the main results are asfollows:A study on the enzymatic properties of acetylcholinesterase by using Ellman method wasperformed and the kinetics parameters based on Michaelis-Menten mechanism under theconditions of the optimal pH, temperature and time were analyzed. It was found thatinhibition rate of acetylcholinesterase activity by ethyl carbamate was significantly improvedafter the sensibilization using bromine solution. It was found that there was a linearrelationship between inhibition rate of acetylcholinesterase activity and concentration of ethylcarbamate in a certain range of1~7.5mg/L. The linear correlation reached0.9972anddetection limit reached0.04177mg/L. RSD was between1.82%to9.69%.The effect of ethanol on acetylcholinesterase activity was studied, and analysis on thechange of kinetic parameters of acetylcholinesterase in ethanol-water and ethanol-buffersolution was performed. In ethanol-buffer system which ethanol concentration was less than15%, there was a linear relationship between ethyl carbamate concentration and its inhibitionrate on enzyme activity, and the linearity range was between1~5mg/L and2~10mg/L.PDAB method analysis revealed that the absorbance can be determined under theconditions of the optimal acid and chromogenic agent, temperature and time. There was alinear relationship between the absorbance and the concentration of ethyl carbamate, and thelinearity range was between0~1g/L, and the method was with high precision. Combination ofPDAB method with enzyme inhibition method, showed that there was no inhibition onacetylcholinesterase by ethyl carbamate after Ehrlich reaction.The investigation on ethyl carbamate level of alcoholic beverages in the mid to low-endmarket of Guangzhou showed that there was a certain amount of ethyl carbamate in variouskinds of alcoholic beverages besides of beer and rice wine, the level was randed as followingbased on their concentration: Brandy> plum wine> yellow rice wine> white wine> medicinalwine> fruit wine> wine. Rapid detection of ethyl carbamate in alcoholic beverages by enzymeinhibition method was conducted. It’s found that the direct application of enzyme inhibitionmethod showed not feasible. Solid phase extraction combined with enzyme inhibition methodcould be only used to qualitative analysis.