Chemical Microdetermination of Heparin in Plasma |
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| Institution: | 1. Faculty of Pharmaceutical Sciences, Chiba University, Yayoi, Inage, Chiba, 263, Japan;2. Division of Medicinal and Natural Products Chemistry and Department of Chemical and Biochemical Engineering, PHAR S328, University of Iowa, Iowa City, Iowa, 52242;1. Department of Pharmaceutical Sciences, University of Perugia, Via del Liceo 1, 06123 Perugia, Italy;2. Department of Chemistry, Biology and Biotechnology, University of Perugia, Via del Giochetto, 06126 Perugia, Italy;3. Department of Clinical and Experimental Medicine, Medical School, University of Foggia, viale Luigi Pinto, 1, 71100 Foggia, Italy;4. Department of Biochemical Sciences, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy;5. Laboratory of Virology – Regina Elena Cancer Institute, Via delle Messi d’Oro 156, 00158 Rome, Italy;1. Beijing National Laboratory for Molecular Sciences, Key Laboratory of Engineering Plastics, Institute of Chemistry, Chinese Academy of Sciences, Zhongguancun North First Street 2, Beijing 100190, PR China;2. University of Chinese Academy of Sciences, Beijing 100049, PR China;1. Spectral Service AG, Emil-Hoffmann-Straße 33, 50996 Cologne, Germany;2. Institute of Chemistry, Saratov State University,Astrakhanskaya Street 83, 410012 Saratov, Russia;3. Institute of Chemistry, Saint Petersburg State University, 13 B Universitetskaya Emb., St. Petersburg 199034, Russia;4. Hemostasis and Thrombosis, Department of Pathology, Loyola University Medical Center, Maywood, IL, United States;5. Ronnsi Pharma Co., LTD, Suite 328, Bldg. A2, bioBay 218 Xinghu, Rd/SIP, JiangSu 215123, China;1. Department of Radiology, Mayo Clinic, United States;2. Thoracic Disease Research Unit, Mayo Clinic, United States;3. Department of Pulmonary and Critical Care Medicine, Mayo Clinic, United States |
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| Abstract: | A new, simple, and highly sensitive method for the determination of heparin has been established. Heparin was first converted into unsaturated disaccharides through the action of heparin lyases I, II, and III. A major trisulfated unsaturated disaccharide product results, consistent with structural analysis of a number of pharmaceutical heparins using one- and two-dimensional1H NMR spectroscopy. This disaccharide was analyzed by HPLC using fluorometric postcolumn derivatization. The correlation between the amount of this trisulfated unsaturated disaccharide and anticoagulant activity of heparin as measured by anti-IIa was determined. The analysis of these pharmaceutical heparins showed a linear correlation between both HPLC and bioassay. This HPLC method was then applied to a pharmacokinetic study of heparin intravenously administered to rabbits. |
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