Simultaneous determination of proline and pipemidic acid in human urine by capillary electrophoresis with electrochemiluminescence detection.

Pipemidic acid is extensively used in the treatment of Gram-negative urinary tract infections, and the contents of proline in human urine vary in association with chronic uremia. The simultaneous determination of pipemidic acid and proline in human urine is of significance for quality control of the dosage and clinical study. The coupling of Ru(bpy)(3)(2+)-based electrochemiluminescence detection with capillary electrophoresis was developed for the simultaneous determination of proline and pipemidic acid in human urine. Parameters related to the separation and detection were investigated and optimized. The standard curves were linear between 0.1 and 90 µg mL(-1) for proline and between 0.4 and 100 µg mL(-1) for pipemidic acid. Underoptimized conditions, the detection limits (3σ) were 0.02 µg mL(-1) for proline and 0.06 µg mL(-1) for pipemidic acid. Relative standard derivations for the electrochemiluminescence intensity and the migration time were 3.2 and 0.9% for proline and 3.7 and 1.2% for pipemidic acid, respectively. The developed method was successfully applied to determine proline and pipemidic acid in human urine. The result showed that the content and decreasing rates of proline in urine for male were higher than that for female, and the content and decreasing rate of pipemidic acid in urine for male and female were consistent, respectively. J. Clin. Lab. Anal. 24:327-333, 2010.