Objectives: Ingested ketogenic agents offer the potential to enhance endurance performance via the provision of an alternative exogenous, metabolically efficient, glycogen-sparing fuel (i.e. ketone bodies). This study aimed to assess the impact of combined carbohydrate and 1,3-butanediol (CHO-BD) supplementation on endurance performance, blood beta-hydroxybutyrate (βHB) concentration and glycolytic activity, in comparison to carbohydrate supplementation alone (CHO). Design: Eleven male runners (age 38 ± 12 years, mass 67.3 ± 6.5 kg, height 174.5 ± 5.0 cm, V O2peak 64.2 ± 5.0 ml•kg-1 •min-1) performed two experimental trials in a randomised crossover design. Methods: Each trial consisted of 60 min of submaximal running, followed by a 5 km running time-trial (TT), and was performed following the ingestion of an energy matched ~650 ml drink (CHO-BD or CHO). Results: There was no difference in TT completion time between the trials (CHO: 1265 ± 93, CHO-BD: 1261 ± 96 s; p=0.723). However, blood βHB concentration in the CHO-BD trial was at least double that of the CHO trial at all time points following supplementation (p<0.05). While blood lactate concentration was lower in the CHO-BD versus CHO trial after 30 min submaximal exercise (CHO-BD: 1.46 ± 0.67 mmol•L-1 , CHO: 1.77 ± 0.46 mmol•L-1 , p=0.040), it was similar at other time points. Blood glucose concentrations were higher post-TT in the CHO-BD trial (CHO-BD: 5.83 ± 1.02 mmol•L-1 , CHO: 5.26 ± 0.95 mmol•L-1 , p=0.015). Conclusions: An energy matched CHO-BD supplementation drink raised βHB concentration and acutely lowered blood lactate concentration, without enhancing 5km TT running performance.