The Italian Decree on Health and Safety at Work (81/08) prescribes mandatory drug tests for jobs which pose safety hazards to others. Workplace drug testing is performed in accordance with the Provision of the Government-Regions Conference, 2008. The aim of our survey was to examine the prevalence of drug use and the main drug findings in a sample of Italian workers performing hazardous jobs. From September 2009 to February 2011, 551 urine samples were collected in 42 Italian companies. Sample collection was carried out at the workplace by qualified laboratory personnel sent from the Institute of Occupational Medicine of the Catholic University (UCSC) of Rome. The workers to be tested were informed the day before, as the law requires. The samples were checked for adulteration, coded, and sent immediately to the laboratory of the UCSC Forensic Toxicology Analytical Unit. The screening test was an immunoassay. The positive samples proceeded to the confirmatory analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The urine samples were analyzed for cannabis, opiates, amphetamines, methamphetamines, cocaine, methadone, and MDMA. Out of 16 samples .9% screened positive; only 4 of them (0.7%) were confirmed with the LC-MS/MS. Confirmed results included cocaine (2 samples), cannabis (1 sample), both cocaine and cannabis (1 sample). The prevalence of positive samples was lower than expected. Such finding cannot be explained by a low reliability of the testing procedure but could be due to test scheduling. More positive cases might be found performing short-notice random testing.