Abstract
Nitrite and nitrate are widely used reporters of endogenous nitric oxide (NO) and nitric oxide synthase (NOS) activity, which are crucial for a broad spectrum of physiological and pathophysiological pathways. Because of the great variety in spatial expression and activity of NOS in animal tissues, a high-resolution analysis of nitrite/nitrate concentrations in very small biological samples, such as individual cells or homogeneous cell clusters, is required. A high-performance capillary zone electrophoresis (CZE) system, which includes a PrinCE-476 computerized capillary electrophoresis and Crystal-1000 conductivity detector, was optimized to analyze nitrite/nitrate concentrations in submicroliter samples of mammalian neuronal tissues and large individual cells of invertebrates. Solid-phase microextraction (SPME) and Isotachophoretic stacking (ITS) were used. The method is highly reproducible and yield excellent limits of detection (LODs): 8.9 nM (0.41 ppb) and 3.54 nM (0.22 ppb) for nitrite and nitrate, respectively, relative to undiluted samples.
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Boudko, D.Y. (2004). High-Resolution Capillary Electrophoresis of Nitrite and Nitrate in Biological Samples. In: Hassid, A. (eds) Nitric Oxide Protocols. Methods in Molecular Biology™, vol 279. Humana Press. https://doi.org/10.1385/1-59259-807-2:009
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DOI: https://doi.org/10.1385/1-59259-807-2:009
Publisher Name: Humana Press
Print ISBN: 978-1-58829-237-7
Online ISBN: 978-1-59259-807-6
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