MP23:Development of a Microfluidic-Based Screening Device for the Detection of Methylarginines in Plasma

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Thomas Linz, University of Kansas

The methylated arginines (MAs) monomethylarginine (MMA), asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) have been shown to be independent predictors of cardiovascular disease. Because these biomarkers could potentially track disease onset and progression, an analytical method capable of quantifying MAs in human plasma has been developed utilizing capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection. The effects of borate concentration, sulfobutylether-b-cyclodextrin concentration, pH and voltage were evaluated to determine the optimal CE separation conditions. Baseline resolution was achieved between naphthalene-2,3-dicarboxaldehyde (NDA)-derivatized arginine, ADMA, MMA and SDMA in less than nine minutes with the optimized method. The limits of detection for these species were experimentally determined to be 5 nM, 20 nM, 40 nM and 20 nM, respectively, which are well below the expected plasma concentrations. MAs from plasma samples were selectively preconcentrated using cation exchange solid phase extraction (SPE) pipette tips and quantified with CE-LIF. The CE separation method was then transferred to a microchip electrophoresis (MCE) device where MAs were baseline resolved in under four minutes on a 14 cm glass microchip. To our knowledge, this is the first report of the separation of MAs by MCE.

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