Caudal epidural analgesia is a well-established therapeutic modality for pain alleviation in horses. Additionally, epidural analgesia could potentially be a complementary diagnostic tool for confirmation of pain-related conditions in horses presenting with nonspecific signs of poor performance or rideability issues. To use the epidural as a diagnostic tool, the administered medications should provide efficient analgesia without accompanying adverse effects. Therefore, the objectives of the current study were to evaluate the analgesic properties and effects on locomotor function, mentation and physical examination parameters of caudal epidural co-administration of methadone and morphine in horses. Five mares received a caudal epidural injection of 0.1 mg/kg bwt methadone and 0.1 mg/kg bwt morphine diluted to a total volume of 4.4 mL/100 kg. Before and several times thereafter, horses were subjected to mechanical nociceptive threshold evaluation, physical examination, assessment of mentation and locomotor function examination. Horses were assigned ataxia scores (0–4) by a group of inexperienced raters (three senior-year veterinary students) and a group of experienced raters (two board-certified internal medicine specialists) that assessed the locomotor examinations either live or video-based. The epidural co-administration of methadone and morphine resulted in clinically relevant and statistically significant increases of horses’ tolerance to mechanical noxious stimuli at the coccygeal, perineal, sacral, lumbar and thoracic regions. Analgesia was evident after 4.4 h and lasted at least 5 h. Regional differences in the onset of analgesia reflected a cranial spread of the analgesic solution. No horses showed signs of gait disturbances; the overall median ataxia score was 0 at all times; and the average difference in scores between two randomly selected raters for a random horse at a random time point was 0.377 indicating high inter-rater agreement. There were no adverse changes of mentation and physical examination parameters. Observed side effects included signs of decreased frequency of defaecation, generalised sweating, and pruritus.