Recently, forensic test for evidence of sweat and other body fluids has increased. However, no plenary test has been developed for demonstration of the presence of sweat, though preliminary tests using urea or lactic acid as indices are widely used. We established a new method for confirming the presence of sweat by high-performance liquid chromatographic (HPLC) detection of two targets, E-urocanic acid (E-UCA) and L-tyrosine (L-Tyr).   Six different body fluid stains (sweat, urine, saliva, semen, vaginal fluid and blood) were extracted with distilled water and the extracts were separated by HPLC on a ODS column. The mobile phase was 50 mM phosphate buffer (pH 2.2) and the UV detector was set at 231 nm. The column temperature was maintained at 40°C.   Two targets were detected in the sweat stains, but E-UCA was not detected in other body fluid stains. These results indicate that the presence of E-UCA is specific for sweat. This method was capable of identifying sweat stains on samples that had been stored for 6 months at room temperature, and should prove very useful for the identification of sweat stains in forensic cases.