Evaluation of mobile phase, ion pairing, and temperature influence on an HILIC-MS/MS method for L-arginine and its dimethylated derivatives detection.
Asymmetric N(G),-N(G)-dimethylarginine (ADMA) increases in diseases such as renal failure, diabetes mellitus, and hypercholesterolemia. The feasibility and utility of a hydrophilic interaction chromatography (HILIC) method for the separation of free L-arginine (Arg), ADMA, and symmetric N(G),-N(G')-dimethylarginine (SDMA) on a typical silica column were explored and the impact of some experimental parameters on the chromatographic behavior of these analytes was investigated. The effect of water and TFA content in mobile phase and of column temperature was investigated during the development of a fast and simple HILIC-MS/MS method that might be suitable for the quantification of free Arg, ADMA, and SDMA in plasma for routine analysis. Our results show that a good compromise between efficiency and peak shape with acceptable retention and total chromatographic run time is achieved using an ACN/water (90:10) mobile phase with TFA% as additive ranging from 0.015 to 0.025% and column temperature ranging from 25 to 30 degrees C.
Paglia
Giuseppe
G
Clinical Biochemistry, Department of Biomedical Sciences, University of Foggia, Foggia, Italy.
D'Apolito
Oceania
O
Tricarico
Filomena
F
Garofalo
Daniela
D
Corso
Gaetano
G
D000181:Acrylonitrile; D001120:Arginine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000592:standards; D001774:Blood Chemical Analysis / Q000379:methods / Q000592:standards; D002853:Chromatography, Liquid / Q000379:methods*; D007202:Indicators and Reagents; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature; D014269:Trifluoroacetic Acid
D007202:Indicators and Reagents; C024917:symmetric dimethylarginine; C018524:N,N-dimethylarginine; D001120:Arginine; D014269:Trifluoroacetic Acid; D000181:Acrylonitrile