Development of a high-throughput differential mobility separation–tandem mass spectrometry (DMS-MS/MS) method for clinical urine drug testing

•A sensitive and specific DMS-MS/MS method is described for simultaneous screening of 33 drugs and metabolites in urine.•The method achieved limits of detection ranging from 5 to 100 ng/mL.•The method was validated and used to screen 56 clinical urine samples.•Results were compared with previously obtained urine toxicology immunoassay results.•DMS-MS/MS was more sensitive and identified 20% more drugs in urine than immunoassay.