Synthesis and Application of MUGal which is the Specific Fluorescent Substrate for Detection of Coliform
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Abstract:
4-Methylumbelliferyl-β-D-galactopyranoside (MUGal) is specific fluorescent substrate for rapid detection of coliform bacteria which are the important indicator bacteria for food safety. Protected glycoside was produced from condensation of 4-methylumbelliferone (4-MU) and tetraacetyl-α-D-galactopyranosyl bromide under the conditions of phase transfer catalysis, and then deprotection in H2O/CH3OH solution of sodium carbonate gave 4-methylumbelliferyl-β-D-galactopyranoside. Through the study of temperature, time and amount of catalyst on the yield of glycosylation，we found that when the temperature was 30 ℃ and the reaction time was 16 h, the yield could be up to 73.12%. Yield of alcoholysis reaction with sodium carbonate as base was 91.01%. Structural characterization of target compound was conducted by IR and 1H NMR. The product was further applied to coliform chromogenic medium. Compared with imported products, they were no significant differences in the number of fluorescent tubes by X2 test (P>0.05). In sample test, both positive rate of GB4789.3-2010 (48 h) and MUGal(24 h) were 80%. The detection time was significantly shorter than GB 4789.3-2010. Therefore, the product could be comparable with imported products and could be used as a substitute for imported products.
