Simultaneous determination of citrulline and arginine in human blood plasma by capillary electrophoresis with ultraviolet absorption detection

Abstract

A new capillary electrophoresis method to measure human blood plasma arginine and citrulline levels in a single run without derivatization was established. After adding homoarginine as internal standard, plasma proteins were removed by a 90:10 v/v acetonitrile/ammonia mixture. Arginine and citrulline were detected by an ultraviolet detector at 190 nm and separated in 11.65 and 20.43 min, respectively, by using a 75 mmol/L Tris phosphate solution at pH 1.2 as a background electrolyte. Limits of detection were 0.8 and 5 μmol/L for arginine and citrulline, respectively. Precision tests indicated a good repeatability of migration times and of peak area both for citrulline (CV% = 0.82 and 3.19) and arginine (CV% = 0.65 and 2.79). The CV% for intra- and interassay tests were, respectively, 1.84 and 3.23 for citrulline and 1.25 and 1.50 for arginine. Mean recovery was 101.5 and 98.5% for citrulline and arginine, respectively. The performance of the developed method was assessed by measuring plasma arginine levels in 52 subjects and the data were compared with those obtained by our previous assay. The new method was then applied to assess plasma citrulline and arginine in ten chronic kidney disease patients under hypolipidemic therapy with statin.

Keywords: Arginine; Capillary electrophoresis; Chronic kidney disease; Citrulline.