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Licensed Unlicensed Requires Authentication Published by De Gruyter June 1, 2005

Determination of asymmetric dimethylarginine (ADMA) using a novel ELISA assay

  • Friedrich Schulze , Reinhard Wesemann , Edzard Schwedhelm , Karsten Sydow , Jennifer Albsmeier , John P. Cooke and Rainer H. Böger

Abstract

Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of nitric oxide synthase (NOS). Elevated ADMA plasma levels have been reported in connection with diseases associated with an impaired endothelial L-arginine-NO pathway and endothelial dysfunction, such as atherosclerosis, hypercholesterolemia, chronic heart failure, diabetes mellitus, and hypertension. NO production by NOS is decreased due to elevated ADMA levels. In fact, there is increasing interest in determination of ADMA levels in samples of various origins. The aim of this work was to develop a precise and easy immunoassay in contrast to the existing methods, such as HPLC, liquid chromatography-mass spectrometry (LC-MS) and gas chromatography (GC)-MS. We determined cross-reactivity in our immunoassay of 1.2% for symmetric dimethylarginine and <0.02% for L-arginine. The limit of quantitation was 0.05μmol/l. We found good correlation of the values measured when we compared our assay with LC-tandem MS (n=29; r=0.984; p<0.0001). We determined ADMA levels in human serum and plasma, mouse and rat plasma, and cell culture supernatant. For human plasma we found a mean of 0.65μmol/l in healthy subjects. In the plasma of mice and rats we found mean concentrations of 1.05 and 1.09μmol/l, respectively.


Corresponding author: Prof. Dr. med. Rainer H. Böger, Arbeitsgruppe Klinische Pharmakologie, Institut für Experimentelle und Klinische Pharmakologie, Universitätsklinikum Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany Phone: +40-42803-9759, Fax: +40-42803-9757,

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Received: 2004-8-3
Accepted: 2004-9-10
Published Online: 2005-6-1
Published in Print: 2004-12-1

©2004 by Walter de Gruyter Berlin New York

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