Evaluation of mobile phase, ion pairing, and temperature influence on an HILIC‐MS/MS method for L‐arginine and its dimethylated derivatives detection

Abstract

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.

Application of amino acid analysis using hydrophilic interaction liquid chromatography coupled with isotope dilution mass spectrometry for peptide and protein quantification

“…A similar drastic change of the retention time was observed when increasing the trifluoroacetic acid (TFA) percentage in the mobile phase [20], which suggested that strong negative charges might interfere with the HILIC mechanism.…”

Quantification of l-arginine, asymmetric dimethylarginine and symmetric dimethylarginine in human plasma: A step improvement in precision by stable isotope dilution mass spectrometry

“…However, ARG, ADMA and SDMA are suitable analytes for the nowadays well established hydrophilic interaction (HILIC) chromatography [16], which is performed on a polar stationary phase with a mobile phase rich in organic modifier. As it turned out, a certain amount of trifluoroacetic acid in the mobile phase was necessary to produce tailing free peaks of the analytes [17]. Since it is well known that trifluoroacetic acid in the mobile phase causes signal suppression in an ESI ion source, a weaker acid (in this case propionic acid) had to be added in excess of the trifluoroacetic acid to alleviate this effect [18].…”

Investigation on performance of zirconia and magnesia–zirconia stationary phases in hydrophilic interaction chromatography

“…Column temperature is an important factor in HILIC separations [27][28][29] as it influences significantly on the following properties: (i) diffusivity of the analytes; (ii) viscosity of the MP, which is closely correlated with hydrophobicity and hydrophilicity of the MP; (iii) enthalpy and entropy change of the analytes in the transferring process from the MP to SP; (iv) pKa of the analytes and pH of the MP [30][31][32]. An increasing number of reports [18][19][20]25,[27][28][29]31,[33][34][35][36][37] on thermodynamics in HILIC were found in recent years, which provided necessary theory guidance when column temperature was used as a variant regulating selectivity and analysis speed.…”