High-throughput liquid chromatographic-tandem mass spectrometric determination of arginine and dimethylated arginine derivatives in human and mouse plasma

The balance between nitric oxide (NO) and vasoconstrictors like endothelin is essential for vascular tone and endothelial function. l-Arginine is converted to NO and l-citrulline by NO synthase (NOS). Asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) are endogenous inhibitors of NO formation. ADMA is degraded by dimethylamino dimethylhydrolases (DDAHs), while SDMA is exclusively eliminated by the kidney. In the present article we report a LC-tandem MS method for the simultaneous determination of arginine, ADMA, and SDMA in plasma. This method is designed for high sample throughput of only 20-μl aliquots of human or mouse plasma. The analysis time is reduced to 1.6 min by LC-tandem MS electrospray ionisation (ESI) in the positive mode. The mean plasma levels of l-arginine, ADMA, and SDMA were 74 ± 19 (SD), 0.46 ± 0.09, and 0.37 ± 0.07 μM in healthy humans (n = 85), respectively, and 44 ± 14, 0.72 ± 0.23, and 0.19 ± 0.06 μM in C57BL/6 mice. Also, the molar ratios of arginine to ADMA were different in man and mice, i.e. 166 ± 50 and 85 ± 22, respectively.