Abstract
Microchip capillary electrophoresis (MCE) is a relatively new analytical method requiring only small sample amounts, which is very favorable for the analysis of volume-limited biofluids. The practical use of MCE in bioanalysis is still restricted in terms of requirements for simplifying and/or concentrating sample pretreatment techniques. Here, we describe an MCE method for trace analysis of nitrite and nitrate, indicators of various neurological diseases, in cerebrospinal fluid (CSF). The complex CSF samples were simplified by solid-phase microextraction prior to an online combination of isotachophoresis with capillary zone electrophoresis performed on a microchip with coupled channels and a high-volume sample injection channel (9.9 μL). The method is suitable for rapid (total analysis time lasted 20 min), reproducible (0.6–2.4 % RSD for migration time), and sensitive (3–9 nM limits of detection) determinations of nitrite and nitrate in 15–50 times diluted CSF samples.
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Esplugues JV (2002) NO as a signalling molecule in the nervous system. Br J Pharmacol 135:1079–1095
Kelm M (1999) Nitric oxide metabolism and breakdown. Biochim Biophys Acta 1411:273–289
Feelisch M, Stamler JS (1996) Measurements of NO-related activities – which assay for which purpose. In: Feelisch M, Stamler J (eds) Methods in nitric oxide research. Wiley, Chichester, pp 303–309
Steinberg A, Wiklund NP, Brundin L et al (2010) Levels of nitric oxide metabolites in cerebrospinal fluid in cluster headache. Cephalalgia 30:696–702
Danilov AI, Andersson M, Bavand N et al (2003) Nitric oxide metabolite determinations reveal continuous inflammation in multiple sclerosis. J Neuroimmunol 136:112–118
Murawska-Ciałowicz E, Szychowska Z, Trebusiewicz B (2000) Nitric oxide production during bacterial and viral meningitis in children. Int J Clin Lab Res 30:127–131
Kuiper MA, Visser JJ, Bergmans PLM et al (1994) Decreased cerebrospinal fluid nitrate levels in Parkinson's disease, Alzheimer’s disease and multiple system atrophy patients. J Neurol Sci 121:46–49
Henry CS (2006) Microchip capillary electrophoresis: methods and protocols. Humana, Totowa
Manz A, Eijkel JCT (2001) Miniaturization and chip technology. What can we expect? Pure Appl Chem 73:1555–1561
Kaniansky D, Masár M, Bodor R et al (2003) Electrophoretic separations on chips with hydrodynamically closed separation systems. Electrophoresis 24:2208–2277
Miyado T, Wakida S-i, Aizawa H et al (2008) High-throughput assay of nitric oxide metabolites in human plasma without deproteinization by lab-on-a-chip electrophoresis using a zwitterionic additive. J Chromatogr A 1206: 41–44
Miyado T, Tanaka Y, Nagai H et al (2004) Simultaneous determination of nitrate and nitrite in biological fluids by capillary electrophoresis and preliminary study on their determination by microchip capillary electrophoresis. J Chromatogr A 1051:185–191
Miyado T, Tanaka Y, Nagai H et al (2006) High-throughput nitric oxide assay in biological fluids using microchip capillary electrophoresis. J Chromatogr A 1109:174–178
Troška P, Chudoba R, Danč L et al (2013) Determination of nitrite and nitrate in cerebrospinal fluid by microchip electrophoresis with microsolid phase extraction pre-treatment. J Chromatogr B 930:41–47
Foret F, Křivánková L, Boček P (1993) Capillary zone electrophoresis. VCH, Weinheim
Bodor R, Kaniansky D, Masár M et al (2003) Determination of bromate in drinking water by zone electrophoresis-isotachophoresis on a column-coupling chip with conductivity detection. Electrophoresis 23:3630–3637
Boudko DY, Cooper BY, Harvey WR et al (2002) High-resolution microanalysis of nitrite and nitrate in neuronal tissues by capillary electrophoresis with conductivity detection. J Chromatogr B 774:97–104
Grass B, Neyer A, Jöhnck M et al (2001) A new PMMA-microchip device for isotachophoresis with integrated conductivity detector. Sensor Actuator B Chem 72:249–258
Everaerts FM, Beckers JL, Verheggen TPEM (1976) Isotachophoresis: theory, instrumentation and applications. Elsevier, Amsterdam
Acknowledgments
This work was supported by the Slovak Grant Agency for Science (VEGA 1/1149/12) and by the Slovak Research and Development Agency (APVV-0259-12). Cerebrospinal fluid samples were provided by Prof. Turčáni (first neurological clinic of Medical Faculty and University Hospital, Comenius University in Bratislava). The financial support of Merck (Darmstadt, Germany) is also acknowledged.
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Masár, M., Bodor, R., Troška, P. (2015). Microchip Capillary Electrophoresis of Nitrite and Nitrate in Cerebrospinal Fluid. In: Van Schepdael, A. (eds) Microchip Capillary Electrophoresis Protocols. Methods in Molecular Biology, vol 1274. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2353-3_3
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DOI: https://doi.org/10.1007/978-1-4939-2353-3_3
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