High-performance liquid chromatographic assay of asymmetric dimethylarginine, symmetric dimethylarginine, and arginine in human plasma by derivatization with naphthalene-2,3-dicarboxaldehyde

Marra, Maurizio; Bonfigli, Anna Rita; Testa, Roberto; Testa, I; Gambini, Anna; Coppa, Gilberto

doi:10.1016/s0003-2697(03)00157-x

Cited by 62 publications

(50 citation statements)

References 30 publications

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“…The mobile phase consisted of buffer A (50 mM KH 2 PO 4 , pH 7.0) and buffer B (ACN/MeOH 70:30) run at room temperature with a flow rate of 1.3 ml/min. The following gradient method was used: 0 -10 min with 90% A, and then 10 -40 min with a linear gradient from 90% A to 30% A (22,39). Intracellular levels of L-Arg and methylarginines were determined from values derived from standard curves of each analyte using the ESA peak integration software assuming intracellular water content of 2 pl (40).…”

Section: Methodsmentioning

confidence: 99%

Evidence for the Pathophysiological Role of Endogenous Methylarginines in Regulation of Endothelial NO Production and Vascular Function

Cardounel

Cui

Samouilov

et al. 2007

Journal of Biological Chemistry

225

172

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In endothelium, NO is derived from endothelial NO synthase (eNOS)-mediated L-arginine oxidation. Endogenous guanidinomethylated arginines (MAs), including asymmetric dimethylarginine (ADMA) and N G -methyl-L-arginine (L-NMMA), are released in cells upon protein degradation and are competitive inhibitors of eNOS. However, it is unknown whether intracellular MA concentrations reach levels sufficient to regulate endothelial NO production. Therefore, the dose-dependent effects of ADMA and L-NMMA on eNOS function were determined. Kinetic studies demonstrated that the K m for L-arginine is 3.14 M with a V max of 0.14 mol mg The biological significance of guanidino-methylated arginine derivatives has been known since the inhibitory actions of N G -methyl-L-arginine (L-NMMA) 3 on macrophage induced cytotoxicity were first demonstrated. It was subsequently realized that these effects were mediated through inhibition of NO release (1). NO has been demonstrated as a critical effector molecule in the maintenance of vascular function (2-4). In the vasculature, NO is derived from the oxidation of L-Arg, catalyzed by the constitutively expressed enzyme, eNOS (5-7). This endothelial-derived NO diffuses from the vascular endothelium into the smooth muscle cell layer where it activates soluble guanylate cyclase leading to smooth muscle relaxation (2-4). In addition to its role in the maintenance of vascular tone, NO helps to maintain the anti-atherogenic character of the normal vascular wall. NO, in concert with various cell signaling molecules, has been demonstrated to maintain smooth muscle cell quiescence and as such, counteracts pro-proliferative agents, specifically those involved in the propagation of athero-proliferative disorders (8 -14). As such, eNOS dysfunction is an early symptom of vascular disease and is manifested through insufficient NO bioavailability. Several potential causes of NO deficiency in disease settings have been proposed. Among these, high circulating levels of the endogenous methylarginine NOS inhibitor asymmetric dimethylarginine (ADMA) has been hypothesized to be of particular importance (15)(16)(17)(18)(19)(20)(21). In neurons and the brain, it has been shown that the methyl arginine L-NMMA is also present, however, the levels of this methylarginine have not been previously considered in studies evaluating vascular dysfunction (22).ADMA and L-NMMA are endogenous inhibitors of NOS and are derived from the proteolysis of methylated arginine residues on various proteins. The methylation is carried out by a group of enzymes referred to as protein-arginine methyltransferase (23). Subsequent proteolysis of proteins containing methylarginine groups leads to the release of free methylarginine into the cytoplasm, and if sufficient levels are reached NO production from NOS would be inhibited (24, 25). To date, six different isoforms of the enzyme have been identified with each

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Section: Methodsmentioning

confidence: 99%

Evidence for the Pathophysiological Role of Endogenous Methylarginines in Regulation of Endothelial NO Production and Vascular Function

Cardounel

Cui

Samouilov

et al. 2007

Journal of Biological Chemistry

225

172

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“…Quantification of ADMA and related metabolites was made by several methods based on HPLC [26][27][28][29][30][31][32], coupled with fluorescence [33] and mass spectrometric methods such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) [34][35][36][37][38], and gas chromatography-mass spectrometry (GC-MS) [39], capillary electrophoresis [40,41] and ELISA [42].…”

Section: Introductionmentioning

confidence: 99%

“…The order of magnitude is different for all metabolites so high specificity and sensitivity is required. GC/MS analysis give good separation of ADMA and SDMA but several extraction and derivatisation steps are required, and sample preparation takes about 2.5-3 h. For HPLC derivatised methods ortho-phthaldialdehyde (OPA), AccQ-Fluor or naphthalene-2,3-dicarboxaldehyde (NDA), are used and analysed by fluorescence [27][28][29][30][31][32][33] or mass spectrometric detection [34]. The crucial points of OPA fluorescence analysis are the rapid decomposition of derivative and the discrimination of ADMA and SDMA only by chromatographic separation.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous quantitative determination of NG,NG-dimethyl-l-arginine or asymmetric dimethylarginine and related pathway's metabolites in biological fluids by ultrahigh-performance liquid chromatography/electrospray ionization-tandem mass spectrometry

Gangi

Chiandetti

Gucciardi

et al. 2010

Analytica Chimica Acta

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“…Current assays for arginine, ADMA and SDMA generally involve ion exchange extraction from plasma followed by HPLC separation with fluorescence detection [8,10,13,14,[17][18][19][20][21], although capillary electrophoresis [1] and liquid chromatography (LC) mass spectrometry [22] have also been described. Derivatisation with ortho-phthaldialdehyde (OPA) and 2-mercaptoethanol is widely used but a major drawback is the instability of the fluorescent derivative necessitating manual or automated derivatisation just prior to analysis by HPLC.…”

Section: Introductionmentioning

confidence: 99%

Determination of l-arginine and NG,NG- and NG,NG′-dimethyl-l-arginine in plasma by liquid chromatography as AccQ-Fluor™ fluorescent derivatives

Heresztyn

Worthley

Horowitz

2004

Journal of Chromatography B

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High-performance liquid chromatographic assay of asymmetric dimethylarginine, symmetric dimethylarginine, and arginine in human plasma by derivatization with naphthalene-2,3-dicarboxaldehyde

Cited by 62 publications

References 30 publications

Evidence for the Pathophysiological Role of Endogenous Methylarginines in Regulation of Endothelial NO Production and Vascular Function

Evidence for the Pathophysiological Role of Endogenous Methylarginines in Regulation of Endothelial NO Production and Vascular Function

Simultaneous quantitative determination of NG,NG-dimethyl-l-arginine or asymmetric dimethylarginine and related pathway's metabolites in biological fluids by ultrahigh-performance liquid chromatography/electrospray ionization-tandem mass spectrometry

Determination of l-arginine and NG,NG- and NG,NG′-dimethyl-l-arginine in plasma by liquid chromatography as AccQ-Fluor™ fluorescent derivatives

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