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5(6)-Carboxy-rhodamine 110 isomers were rapidly synthesized by condensation of 3-aminophenol and trimellitic anhydride using microwave irradiation as heating method. After esterification of the two isomers, separation and hydrolyzation provided target products,5-carboxy-rhodamine 110 and 6-carboxy-rhodamine 110, in high yields. The results shows microwave irradiation can effectively reduce the reaction time and the demanding of condensed H2SO4.