The aim of this study was to evaluate the prevalence and abuse potential of antiepileptic drugs (AEDs) among prison populations in Scotland, UK. Participants consisted of all admitted and released prisoners over a 1month period who consented to provide samples. Urine samples were collected and analyzed by liquid chromatography coupled with triple quadrupole tandem mass spectrometry using a method validated for the simultaneous quantification of 21 AEDs in urine. A total of 904 samples were collected. The samples were also screened for drugs of abuse by using point-of-care testing kits. A total of 18% of the samples were positive for AEDs. Gabapentin (GBP) was identified in 118 samples (13%) and pregabalin (PRG) in 32 samples (3.5%). Interestingly, 12 samples contained both drugs (1.3%). The concentrations ranged from 0.5 to 1,100mg/L (median, 15mg/L) for GBP and from 0.5 to 440mg/L (median, 7.3mg/L) for PRG. Four samples were found to have concentrations >400mg/L, two samples for GBP and two samples for PRG. These concentrations are at least 20 times above the median concentrations. Other AEDs detected were levetiracetam (four samples), vigabatrin (four samples), lamotrigine (three samples), valproic acid (three samples), carbamazepine (two samples) and topiramate (one sample). Illicit or non-prescribed drugs were detected in 81% of urine samples of which 80% were from admitted prisoners and 20% from released prisoners. Benzodiazepines, opiates and cannabis were the most frequently detected drugs. Other drugs found in positive AED samples were methadone (26%), cocaine (18%), buprenorphine (17%), amphetamines (4%), methamphetamines (4%) and barbiturates (4%). This study shows a high prevalence of AEDs within the Scottish prison system, primarily due to GBP and PRG; however, due to the anonymity of the sample collection, it is unknown if these are prescribed or illicit drug ingestions.