Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1213304 | Journal of Chromatography B | 2012 | 4 Pages |
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 13C6-arginine, D7-ADMA and, for the first time in LC–MS/MS, D6-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.
► A quantification method for l-arginine and its metabolites ADMA and SDMA is presented. ► A very easy and fast sample preparation is proposed. ► Stable isotope labeled I.S.(s) are used for each l-arginine, ADMA and SDMA. ► Very good performance regarding precision and accuracy is achieved due to the I.S.(s). ► Matrix independency is achieved due to the I.S.(s).