The current protocol describes an alternative method for creatine quantification in biological tissue samples using capillary electrophoresis, with high separation efficiency that enables rapid analysis (<10 minutes) with low sample volumes. The protocol involves homogenization of snap-frozen tissue samples in phosphate buffer, followed by electrophoresis through a bare-fused capillary (75 µm i.d.) and measurement at 200 nm. Under the optimised conditions, we found excellent linearity in creatine standards between 6.3 – 100 µM. The overall intra-assay variability for concentrations between 6.3 – 100 µM was 1.5 %, and the inter-assay variability was 6.4 %, with a limit of detection at 6 nmol/mg protein. Variables such as the amount of sample injected or the capillary diameter could be changed to enhance the lower limit of detection. The current protocol was developed and optimised using ovine lung tissue samples, but it can be easily adapted to analyse various tissue types.