The direct mass spectrometric determination of highly concentrated analytes in human urine was demonstrated using extractive electrospray ionization without sample dilution or complex preparation. By increasing the distance between the extractive electrospray source and ion inlet of the mass spectrometer from 5 millimeters to 15 centimeters, the fraction of free analyte ions and charged microdroplets introduced into the mass spectrometer was substantially reduced. Consequently, detector saturation, instrument contamination, and space charge effects were greatly diminished for the analysis of highly concentrated samples. Under the optimized experimental conditions, pure aniline and creatinine (>1 millimolar) in human urine were directly characterized by extractive electrospray ionization without any pretreatment. The urinary creatinine concentrations from two adults were 424 ± 30 and 635 ± 32 micrograms per milliliter and were in good agreement with those obtained by a spectrophotometric method based on the Jaffe reaction. The results show that extractive electrospray ionization is suitable for the direct determination of highly concentrated analytes or even pure compounds, allowing rapid characterization of samples in the chemical industry and clinical studies.