Determination of inulin in plasma and urine by reversed-phase high-performance liquid chromatography |
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| Affiliation: | 1. Ghent University, Faculty of Bioscience Engineering, Department of Food Safety and Food Quality, Research Group Food Chemistry and Human Nutrition, Belgium;2. School of Plant and Horticultural Science, Faculty of Agriculture, Hawassa University, P.O. Box 05, Awassa, Ethiopia;1. Graduate Program of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz, km 235, PO Box 676, 13565-905 São Carlos, SP, Brazil;2. Department of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz, km 235, PO Box 676, 13565-905 São Carlos, SP, Brazil;1. University Hospital of Heidelberg, Department of Internal Medicine 1 and Clinical Chemistry, Heidelberg, Germany;2. German Center for Diabetes Research (DZD), Munich-Neuherberg, Germany;3. University Hospital of Heidelberg, Department of Neuroradiology, Heidelberg, Germany;4. NeuroCentrum Odenwald, Darmstadt, Germany;5. Medical Faculty Mannheim, Department of Neurophysiology, University of Heidelberg, Mannheim, Germany;6. Joint Heidelberg-ICD Translational Diabetes Program, Helmoltz-Zentrum, Munich, Germany;1. Department of Food, Agricultural and Biological Engineering, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH, USA;2. Department of Horticulture and Crop Science, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH, USA |
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| Abstract: | We report a new HPLC procedure for measuring inulin in plasma and urine. Samples after dilution are boiled in mild acidic conditions and then analyzed on a C18 column. Solvent system A is 3.2 mM HCl, pH 2.5, and B is acetonitrile-3.2 mM HCl (60:40, v/v), pH 2.5. The separation is carried out in 8 min with a flow-rate of 1.0 ml/min and the absorbance monitored at 280 nm. The relationship between inulin and the recorded peak area is linear from 0.2 to 3.2 mg/ml with a correlation coefficient of 0.999 for plasma and 0.999 for urine. Within-run precision, measured at three inulin concentrations, ranged from 0.9 to 1.7% in plasma and from 0.8 to 1.2% in urine. Between-run precision varied in plasma from 2.7 to 3.2% and in urine from 3.0 to 3.3%. Analytical recovery ranged from 102 to 107% in plasma and from 101 to 105% in urine, respectively. The method is sensitive, selective and only 30-μl samples are required. Therefore, it could be used to evaluate the glomerular filtration rate even in small babies and to perform studies in animals. |
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