Gemifloxacin mesylate is a novel fourth generation fluoroquinolone agent that is used for the treatment of community acquired pneumonia and acute bacterial exacerbation of chronic bronchitis that provide high rates of mortality worldwide. Spectrofluorimetric analysis constitutes a widespread, effective technique to improve analysis selectivity and sensitivity. Moreover, it offers easy and less time-consuming analysis by using simple and available reagents, which are able to be used for routine determinations of drug substances. Therefore, spectrofluorimetric analysis is one of the major interests of analytical pharmacy. Although, limited literatures have been reported for the determination of gemifloxacin spectrofluorometrically. Therefore, the aim of this book was directed for using micelle-enhanced and Hantzsch reaction spectrofluorometric methods for developing new simple, selective and sensitive methods for determination of gemifloxacin in tablets and human plasma. Moreover, it can be effectively used for routine analysis of gemifloxacin in pharmaceutical industries, hospitals and research laboratories.