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Fluorometric measurement of nitrite/nitrate by 2,3-diaminonaphthalene

Nature Protocols volume 1, pages 2223–2226 (2006)Cite this article

Abstract

We describe a step-by-step protocol for measuring the stable products of the nitric oxide (NO) pathway: nitrite, nitrite plus nitrate and nitrate. This described protocol is easy to apply and is about 50 times more sensitive than the commonly used Griess reaction or commercially available assay kits based on the Griess reaction. It also allows the study of minimal changes in the NO pathway. With this method, it takes about 3 h to analyze the above-mentioned stable products in culture supernatants or in various body fluids, and the method has a sensitive linear range of 0.02–10.0 μM. This restricted linear range suggests that the technique is useful for studying small changes of nitrite and nitrate, rather than for routine diagnostic measurements.

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Figure 1
Figure 2: Standard curve for the detection of nitrite in aqueous solutions.

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Acknowledgements

The present work was partially supported by the Federal Ministry of Research (BMBF 0313079A to N.C.N.).

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Authors and Affiliations

  1. Department of General- Visceral- and Transplantation Surgery, University Medicine Berlin, Campus Virchow, Augustenburger Platz 1, Berlin, 13353, Germany

    Andreas K Nussler, Matthias Glanemann, Anja Schirmeier, Liegang Liu & Natascha C Nüssler

  2. Department of Nutrition and Food Hygiene, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, 430030, PR, China

    Liegang Liu

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  1. Andreas K Nussler
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Correspondence to Andreas K Nussler.

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Nussler, A., Glanemann, M., Schirmeier, A. et al. Fluorometric measurement of nitrite/nitrate by 2,3-diaminonaphthalene. Nat Protoc 1, 2223–2226 (2006). https://doi.org/10.1038/nprot.2006.341

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