Quantification of L-arginine, Asymmetric Dimethylarginine and Symmetric Dimethylarginine in Human Plasma: a Step Improvement in Precision by Stable Isotope Dilution Mass Spectrometry

The amino acid L-arginine and its metabolites ADMA and SDMA are important markers for a range of diseases in humans. Increased levels of ADMA and SDMA in plasma point to endothelial dysfunction, hypertension, renal impairment and other pathological states. We present here a method to quantify L-arginine, ADMA and SDMA in human plasma, which is suitable to support clinical research in this field. Sample preparation consisted only of protein precipitation and the analytes were separated using a silica based HILIC column. The analytes were detected by ESI MS/MS, providing high selectivity and sensitivity. The calibration functions were linear in the ranges of 7.5-150 μmol/l for l-arginine, 0.15-3 μmol/l for ADMA and 0.2-4 μmol/l for SDMA. These ranges cover the concentrations encountered in healthy and pathological human plasma. The method employs (13)C(6)-arginine, D(7)-ADMA and, for the first time in LC-MS/MS, D(6)-SDMA as internal standards for L-arginine, ADMA and SDMA. Therefore, matrix independency and a high intra-day precision of 0.82% for L-arginine, 2.12% for ADMA and 2.83% for SDMA, were achieved at basal plasma concentrations. The respective inter-day precision values were 4.01% for l-arginine, 3.77% for ADMA and 3.86% for SDMA.