Analysis of aspartate and glutamate in human cerebrospinal fluid by high-performance liquid chromatography with automated precolumn derivatization |
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| Authors: | D C Spink J W Swann O C Snead R A Waniewski D L Martin |
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| Institution: | 1. Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, New York 12201 USA;2. Department of Pediatrics, University of Alabama in Birmingham, School of Medicine, Birmingham, Alabama 35233 USA;1. October University for Modern Sciences and Arts, Faculty of Pharmacy, Analytical Chemistry Department, 11787 6th October City, Egypt;2. Cairo University, Faculty of Pharmacy, Analytical Chemistry Department, El-Kasr-El Aini St, 11562 Cairo, Egypt;3. School of Life and Medical Sciences, University of Hertfordshire Hosted By Global Academic Foundation, New Capital, Garden City, Cairo R5 New, Egypt;1. Amity Institute of Nanotechnology, Amity University, Noida 201313, UP, India;2. Department of Biochemistry, MD University, Rohtak 124001, HRY, India;1. School of Environmental Science and Public Health, Wenzhou Medical University, Wenzhou 325035, China;2. School of Information and Engineering, Wenzhou Medical University, Wenzhou 325035, China;3. State Key Laboratory of Marine Environmental Science (Xiamen University), College of the Environment and Ecology, Xiamen University, Xiamen 361005, China;1. LIEC-Laboratório Interdisciplinar de Eletroquímica e Cerâmica, Departamento de Química, UFSCar-Universidade Federal de São Carlos, Rod.Washington Luis km 235, CP 676 São Carlos, SP 13565-905, Brazil;2. Instituto de Química de Araraquara, UNESP-Universidade Estadual Paulista, Rua Francisco Degni, CP 355 Araraquara, SP 14801-907, Brazil;1. Natural Resources Institute Finland (Luke), FI-31600, Jokioinen, Finland;2. School of Forensic and Applied Sciences, University of Central Lancashire, Preston, PR1 2HE, United Kingdom;3. Department of Environmental Sciences, University of Helsinki, Niemenkatu 73, FI-15140 Lahti, Finland;1. Department of Allergy and Clinical Immunology, School of Medicine, Kyungpook National University, Daegu, Korea;2. Department of Allergy and Clinical Immunology, Ajou University School of Medicine, Suwon, Korea;3. Office of Hospital Information, Seoul National University Hospital, Seoul, Korea;4. MD Healthcare Inc, Seoul, Korea;5. Department of Computer Science and Engineering, Hanyang University, Seoul, Korea |
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| Abstract: | A method is described for the analysis of the neuroexcitatory amino acids, aspartate and glutamate, in human cerebrospinal fluid (CSF) by reverse-phase, high-performance liquid chromatography. Fluorescent isoindole derivatives of the amino acids were prepared by reacting the amino acids with ortho-phthalaldehyde in an automated, precolumn procedure. Chromatographic conditions were developed that resolve the isoindole derivatives of aspartate and glutamate from those of at least 10 unidentified components of CSF. Amino acids were reliably quantified in 5-microliter samples of CSF, and deproteinization of the specimens was not required. Furthermore, it was found that deproteinization by precipitation with strong acid can lead to artifactually high measurements of glutamate. The concentrations of free aspartate and glutamate in lumbar CSF from 15 neurologically normal children were 0.30 +/- 0.11 and 0.48 +/- 0.26 microM (mean +/- SD), respectively. The value for glutamate is considerably lower than has been reported in any previous study of human CSF. |
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