Background: Substance abuse is a serious problem all over the world. There are many studies report the illegal substance use profile but few studies present their toxicology laboratory analysis. This study reports a quantitative profile of (Urine Drug Screening)) for illegal substances in Sakarya-Turkey.Methods: This study presents the urine analysis of all illegal substances which were made in the laboratory of Sakarya Training Research Hospital between March 2012 and February 2015. The results obtained from socio-demographic data and urine tests of patients were analyzed by examining their hospital record files. Urine drug screening was conducted with immunoassay quantitative analysis.Results: People subjected to substance analysis (n=2948) ages vary between 12 and 76, their mean age was 28.30±9.46. 96.74% (n=2852) of them were males. Substance positivity was determined in 34.73% of all patients (n=1024/2948) and their ages varied between 14 and 70 and their mean age was 29.39±9.65. Distribution of the urine positivity of the substances contained: marijuana 79, 5% (n=814), amphetamine 30.17% (n=309), ecstasy 23.74% (n=199), benzodiazepine 9,1% (n=94), synthetic cannabinoid 4.9% (n=12/243); opioid 5.2% (n=54), cocaine 1.67% (n=14) and multiple substance 29.9% (n=308).Conclusions: According to this study, marijuana is the most frequently used substance and multiple substance use is common. Synthetic cannabinoid seems to take place rapidly among the users. Updating the kits is important to reach the correct information in drug screening tests.