The invention relates to a method of a Mn doped ZnS quantum dots room temperature phosphorescence detecting an enoxacin in the biological body fluids. The invention uses the room temperature phosphorescence properties of the Mn doped ZnS quantum dots, and provides a simple, fast, economical, keen and highly selective method to detect the enoxacin in the biology body fluids; the Mn doped ZnS quantum dots are dissolved to be a solution, a urine sample or a serum sample is diluted into 30, 50 or 80 times for detecting the linear equation of the enoxacin, whose linear range is 0.2-7.2 mu mol / L, and detection limit is 58.6 nmol / L; when the method is used to detect the enoxacin in the biological body fluids, an oxygen scavenger and a inducer are not need to add, and the background fluorescence and the interference of scattered light can be avoided. Simultaneously, due to the high selectivity of the method, when the enoxacin is detected in the biological body fluids, a complex sample pretreatment process is not needed.