The determination of trace elements in human fluids facilitates the identification of metabolic disorders and physiological abnormities in clinical diagnosis. Elemental concentrations in serum, urine, and saliva are often low and highly prone to contamination. Consequently, a simple and ultrasensitive detection technique is required. In this study, a method was developed for the simultaneous determination of trace arsenic and antimony in clinical samples by hydride generation atomic fluorescence spectrometry. The experimental parameters that affect the generation, delivery, and atomization of volatile arsenic and antimony hydrides were investigated. The limits of quantification were calculated to be 0.0037 nanogram per milliliter and 0.0070 nanogram per milliliter for arsenic and antimony, respectively. The method was successfully applied for the analysis of fortified human serum, urine, saliva, and certified reference materials. The results obtained by the reported method were not statistically different from the certified values at the 95 percent confidence level.