title: Determination of asymmetric and symmetric dimethylarginines in human plasma by HPLC with electrochemical detection.
abstract: A new HPLC method was developed and validated for the determination of asymmetric and symmetric dimethylarginines and l-arginine in human plasma. After SPE and evaporation of the eluate, the samples were derivatised with an o-phthaldialdehyde reagent containing 3-mercaptopropionic acid. The derivatives formed were analysed by isocratic RP-HPLC with electrochemical detection at +320 mV. The mobile phase consisted of 50 mM phosphate buffer (pH 6.1) containing 10% v/v acetonitrile, the flow rate was 1 mL/min. The retention times of all compounds including monomethylarginine (internal standard) were <24 min. The LODs (S/N 3:1) were 0.012 μM for both dimethylarginines and 0.013 μM for L-arginine; the linearity of the method was from 0.1 to 20 μM for both dimethylarginines and from 1 to 200 μM for L-arginine. Absolute extraction recoveries measured for all analytes ranged from 85 to 88%.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Masaryk University, Kamenice, Brno, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Kleparnik
authors.forename: Marie
authors.initials: M
authors.lastname: Tomandlova
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Glatz
authors.forename: Josef
authors.initials: J
authors.lastname: Tomandl
mesh_terms: D001120:Arginine / Q000031:analogs & derivatives* / Q000097:blood; D001774:Blood Chemical Analysis / Q000295:instrumentation* / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D055664:Electrochemical Techniques; D006801:Humans; D057230:Limit of Detection; D012015:Reference Standards
chemical_list: C487735:dimethylarginine; D001120:Arginine
keywords: Arginine; Asymmetric dimethylarginine; Electrochemical detection; HPLC; Symmetric dimethylarginine