A simple, sensitive, and selective method for the determination of palmitate has been investigated on the basis of the fluorescence enhancement of Eu3+–TTA complex in the form of Eu3+–TTA–palmitate. An emission peak of Eu3+–TTA, which increased linearly with the addition of palmitate, occurs at 610 nm in aqueous solution with excitation at 306 nm. The linear curve of calibration ranges is from 1.0 × 10−6 to 6.0 × 10−6 M, and the detection limit is 5.0 × 10−7 M. The effects of foreign ions were studied. The method was applied to the determination of palmitate content in synthetic and beef fat samples.