期刊
CLINICA CHIMICA ACTA
卷 492, 期 -, 页码 29-36出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.cca.2019.02.004
关键词
Arginine methylation; Nitric oxide; Diethylpyrocarbonate; Tandem mass spectrometry; Cardiovascular disease
资金
- Ministero dell'Istruzione, dell'Universita e della Ricerca (MIUR, Italy) - Progetti di Ricerca di Rilevante Interesse Nazionale - PRIN 2015 [20157ATSLF_002]
- MIUR-National Research Council (MIUR-CNR, Italy) Flagship InterOmics [PB05]
Background: Changes in NO metabolism correlate with cardiovascular risk factors and are associated with endothelial dysfunction. NO availability is regulated by nitric oxide synthase (NOS) and arginine and some chemically related metabolites and analogs have the capacity to alter NOS activity. Hence the need for analytical methods for the simultaneous assessment of these analytes. Methods: Analytes (L-arginine (Arg), N-G-monomethyl-L-arginine (MMA), L-homoarginine (hArg), asymmetric dimethyl-L-arginine (ADMA), symmetric dimethyl-L-arginine (SDMA), and L-citrulline (CIT)) were isolated from human plasma by thermal coagulation of plasma followed by a derivatization with diethylpyrocarbonate. Carbetoxy derivatives were separated on a C18 reversed-phase column in < 10 min using an aqueous solution of 0.4% v/v formic acid and acetonitrile (95:5, v/v) mixture as a mobile phase. Positive electrospray ionization and tandem mass spectrometry in combination with specific multiple reaction monitoring transitions were used for detection of analytes and three deuterated forms of the analytes used as internal standards. Results: Intra- and inter-day precision %RSD values ranged between 3 and 5.5% and percentage recoveries were close to 100% for all analytes. Plasma concentrations in 20 healthy male volunteers were 58.62 +/- 8.81 mu mol/L for Arg, 105.08 +/- 21.66 nmol/L for MMA, 1.88 +/- 0.57 mu mol/L for hArg, 0.612 +/- 0.140 mu mol/L for ADMA, 0.581 +/- 0.172 mu mol/L for SDMA, and 28.62 +/- 11.60 mu mol/L for Cit, respectively. Conclusion: This LC MS/MS method provides the capacity to quantify the plasma concentrations of arginine and some of its chemically related metabolites. Sample preparation was simple, inexpensive and effortless. Overall, given the short sample preparation and chromatographic run time, the method may be suitable for the fast and reproducible quantitative determination of the analytes in large clinical trials and routine analysis.
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