The technique of isotachophoresis has been used to develop a specific and sensitive method for the determination of citrate in unprocessed urine. The specificity of the isotachophoretic method was assessed using citrate lyase which caused disappearance of the isotachophoretic citrate signal. The isotachophoretic method compared favourably with the enzymatic method (citrate lyase) for urinary citrate. The normal range for urinary citrate in 25 healthy individuals, as found by isotachophoresis, was 0.33–2.89 mmol/24 h with a mean of 2.1 mmol/24 h.