A highly sensitive capillary electrophoresis method using p-nitrophenyl 5′-thymidine monophosphate as a substrate for the monitoring of nucleotide pyrophosphatase/phosphodiesterase activities

► A sensitive nucleotide pyrophosphatase (NPP) assay was developed. ► p-Nitrophenyl 5′-thymidine monophosphate was used as an artificial substrate. ► A CE-VIS method coupled with dynamically polybrene-coated capillaries was used. ► A combination of phosphate-HEPES buffer notably increased the method sensitivity. ► The obtained LOD and LOQ for p-nitrophenolate are in the nanomolar range.