Determination of histidine in human serum and urine by cation exchange chromatography coupled to selective on-line post column derivatization

A reliable and highly selective method for the determination of histidine in human serum and urine is described. Histidine was separated from the matrix by cation exchange chromatography and detected selectively using online post column derivatization and fluorimetric detection. Unique reaction of histidine with o-phthalaldehyde in the absence of nucleophilic compounds offered specific detection in the complex biological substrate. Linearity was obeyed in the range of 0.5 - 25 mu mol L-1 with a limit of detection of 0.160 mu mol L-1. The absence of matrix effect (<5%) enabled the processing of real samples after minimal pretreatment. Endogenous histidine has been determined in human serum in the range of 78 - 119 mu mol L-1 and random human urine in the range of 266 - 2034 mu mol L-1. The percent recoveries were satisfactory in all cases, ranging between 89 and 114%.