Fluorometric determination of streptomycin in serum by high-performance liquid chromatography using mobile phase containing fluorogenic reagent |
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| Authors: | H Kubo H Li Y Kobayashi T Kinoshita |
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| Affiliation: | 1. School of Pharmaceutical Sciences, Kitasato University, 5-9-1, Shirokane, Minato-ku, Tokyo 108, Japan;2. Shenyang College of Pharmacy, 2-7, Wenhua Road, Shenyang, Peoples Republic of China;1. School of Chemistry and Chemical Engineering, Tiangong University, Tianjin, 300387, PR China;2. Tianjin Polytechnic University State Key Laboratory of Separation Membranes and Membrane Processes, Tianjin, 300387, PR China;1. Department of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, Republic of Korea;2. Fuel Cell Research Center, Korea Institute of Science and Technology (KIST), Seoul 136-791, Republic of Korea;3. Department of Materials Science and Engineering, The University of Texas at Dallas, Richardson, Texas 75080, USA;4. WCU Multiscale Mechanical Design Division, School of Mechanical and Aerospace Engineering, Seoul National University, Seoul 151-744, Republic of Korea;5. Graduate School of Knowledge-based Technology and Energy, Korea Polytechnic University, Siheung-si, Gyeonggi-do 429-793, Republic of Korea;1. Laboratory of Pharmaco-Toxicological Analysis, Department of Pharmacy and Biotechnology (FaBiT), Alma Mater Studiorum, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy;2. Inter-Department Centre for Industrial Research – Advanced Mechanics and Materials (CIRI-MAM), Piazza Malatesta 29-30, 47923 Rimini, Italy;3. Institute for the Study of Nanostructured Materials, National Council of Research (ISMN-CNR), Via P. Gobetti 101, 40129 Bologna, Italy;4. Laboratory of Pharmaceutical Botany, Department of Pharmacy and Biotechnology (FaBiT), Alma Mater Studiorum, University of Bologna, Via Irnerio 4, 40126 Bologna, Italy;1. Department of Imaging and Applied Physics, Fuels and Energy Technology Institute, Curtin University, GPO Box U1987, Perth, WA 6845, Australia;2. Interdisciplinary Nanoscience Center (iNANO) and Department of Chemistry, University of Aarhus, DK-800 Aarhus, Denmark;1. State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, College of Materials Science and Engineering, Donghua University, Shanghai 201620, China;2. Indian Institute of Technology Guwahati, Guwahati 781039, India |
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| Abstract: | A new postcolumn derivatization method for the fluorometric determination of streptomycin in serum by high-performance liquid chromatography is described. The serum was treated with 3.5% perchloric acid to precipitate proteins and the supernatant was directly injected into the chromatograph. Streptomycin was separated by reversed-phase, ion-pair chromatography with a mobile phase containing ninhydrin as a fluorogenic reagent, octanesulfonate, and 1,2-ethanedisulfonate as counterions, and was detected by fluorescence using continuous-flow, postcolumn derivatization in an alkaline stream with ninhydrin in the mobile phase. This method is sensitive to 1.0 microgram/ml using only 100 microliter of serum. Comparison with a fluorescence polarization immunoassay gave a good correlation coefficient of 0.976. |
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