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Determination of asymmetric and symmetric dimethylarginines in human plasma by HPLC with electrochemical detection

M. Kleparnik, M. Tomandlova, Z. Glatz, J. Tomandl,

. 2013 ; 36 (23) : 3696-701.

Jazyk angličtina Země Německo

Typ dokumentu časopisecké články, práce podpořená grantem, validační studie

Perzistentní odkaz   https://www.medvik.cz/link/bmc14074606

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%.

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