Synthesis of dihalogenated coumarin derivatives for monitoring acid phosphatase activity

Aim To synthetize 6-chloro-8-fluoro-4-methylumbelliferone phosphate(CF-MUP) and utilize it to analyze acid phosphatase activity.Method CF-MUP obtained by phosphorylation of CFMU(6-chloro-8-fluoro-4-methylumbelliferone) which was prepared by Pechmann reaction between 4 and ethyl acetoacetate in the presence of methanesulfonic acid.And then,the utility of CF-MUP as a substrate for acid phosphatase was compared with MUP(4-methylumbelliferone phosphate).Results The new coumarin phosphate,CF-MUP,demonstrated much higher sensitivity and was more robust for detecting the activity of acid phosphatase than MUP in the physiological pH range.Conclusion This new fluorescence dye,CF-MUP,is a convenient tool for the continuous assay of acid phosphatases with a wide range of pH.