Resolution and quantification of arginine, monomethylarginine, asymmetric dimethylarginine, and symmetric dimethylarginine in plasma using HPLC with internal calibration
| العنوان: | Resolution and quantification of arginine, monomethylarginine, asymmetric dimethylarginine, and symmetric dimethylarginine in plasma using HPLC with internal calibration |
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| المؤلفون: | Glenn Nardone, Matthew S. Alkaitis, Jessica H. Chertow, Hans Ackerman |
| المصدر: | Biomedical Chromatography |
| بيانات النشر: | Wiley, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | Male, 0301 basic medicine, Methylarginine, Vascular Homeostasis, Arginine, Coefficient of variation, Clinical Biochemistry, 01 natural sciences, Biochemistry, High-performance liquid chromatography, Analytical Chemistry, Mice, 03 medical and health sciences, chemistry.chemical_compound, Limit of Detection, SDMA, Drug Discovery, Animals, Humans, L-NMMA, Molecular Biology, Research Articles, Chromatography, High Pressure Liquid, Pharmacology, Chromatography, Arginine transport, biology, 010401 analytical chemistry, Reproducibility of Results, General Medicine, 0104 chemical sciences, ADMA, Mice, Inbred C57BL, Standard curve, Nitric oxide synthase, 030104 developmental biology, chemistry, Calibration, Linear Models, biology.protein, Nitric Oxide Synthase, Asymmetric dimethylarginine, Research Article |
| الوصف: | NG,NG‐dimethyl‐l‐arginine (asymmetric dimethylarginine, ADMA),NG‐monomethyl‐l‐arginine (l‐NMMA) and NG,N G’‐dimethyl‐l‐arginine (symmetric dimethylarginine, SDMA) are released during hydrolysis of proteins containing methylated arginine residues. ADMA and l‐NMMA inhibit nitric oxide synthase by competing with l‐arginine substrate. All three methylarginine derivatives also inhibit arginine transport. To enable investigation of methylarginines in diseases involving impaired nitric oxide synthesis, we developed a high‐performance liquid chromatography (HPLC) assay to simultaneously quantify arginine, ADMA, l‐NMMA and SDMA. Our assay requires 12 μL of plasma and is ideal for applications where sample availability is limited. We extracted arginine and methylarginines with mixed‐mode cation‐exchange columns, using synthetic monoethyl‐l‐arginine as an internal standard. Metabolites were derivatized with ortho‐phthaldialdeyhde and 3‐mercaptopropionic acid, separated by reverse‐phase HPLC and quantified with fluorescence detection. Standard curve linearity was ≥0.9995 for all metabolites. Inter‐day coefficient of variation (CV) values were ≤5% for arginine, ADMA and SDMA in human plasma and for arginine and ADMA in mouse plasma. The CV value for l‐NMMA was higher in human (10.4%) and mouse (15.8%) plasma because concentrations were substantially lower than ADMA and SDMA. This assay provides unique advantages of small sample volume requirements, excellent separation of target metabolites from contaminants and validation for both human and mouse plasma samples. © 2015 The Authors Biomedical Chromatography published by John Wiley & Sons, Ltd. |
| تدمد: | 1099-0801 0269-3879 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ac15bacd636332a22f4c28823362ba8eTest https://doi.org/10.1002/bmc.3548Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....ac15bacd636332a22f4c28823362ba8e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10990801 02693879 |
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