Determination of nitrite in human blood by combination of a specific sample preparation with high-performance anion-exchange chromatography and electrochemical detection

Journal of Chromatography. B, Biomedical Applications
H Preik-Steinhoff, M Kelm

Abstract

All photometric or HPLC methods described to date have been unable to detect nitrite, a reliable marker of NO synthase activity, in human blood because of its rapid metabolism within the erythrocytes. We now elaborate on method to prevent nitrite degradation during sample preparation which in combination with high-performance anion-exchange chromatography and electrochemical detection allows a sensitive measurement of nitrite. A linear current response in the concentration range of 10-1000 nmol/l nitrite was observed yielding a correlation coefficient of 0.99. In addition, the combination of the electrochemical with a UV detector allowed us to simultaneously quantify nitrate within one analytical run, which is the end product of NO/nitrite metabolism. Basal levels for nitrate and nitrite in human blood were determined with 25 +/- 4 mumol/l and 578 +/- 116 nmol/l (n = 8), respectively and thus were in the same concentration range as expected from NO measurement in saline perfused isolated organs or cultured endothelial cells. Therefore, the presented method may be used to assess activity of endothelial constitutive NO synthase in humans under physiological and pathophysiological conditions.

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Citations

Nov 1, 2012·Bulletin of Experimental Biology and Medicine·V Yu TitovA N Osipov
May 4, 2004·Blood Cells, Molecules & Diseases·André DejamMark T Gladwin
May 13, 1999·Biochimica Et Biophysica Acta·M Kelm
Jul 1, 1998·Clinical Biochemistry·G EllisS K Makela
Nov 3, 2001·Journal of the American College of Cardiology·Y IshibashiM Hashimoto
Jun 8, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Dmitri Y BoudkoLeonid L Moroz
Oct 4, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Christopher C T SmithD John Betteridge
Oct 18, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Vera JedlickováStefan Alusík
Mar 22, 2002·Clinical and Experimental Pharmacology & Physiology·Takashi SugamoriYo Murakami
Nov 19, 2004·Antioxidants & Redox Signaling·Paul W Buehler, Abdu I Alayash
Jan 13, 2004·Journal of Cardiovascular Pharmacology·Stefan Martin PeringsMalte Kelm
Nov 2, 2007·Clinical and Experimental Pharmacology & Physiology·Darren P CaseyRandy W Braith
Feb 14, 2004·Redox Report : Communications in Free Radical Research·Joseph M RifkindLuke Babu Ravi
Oct 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·T LauerM Kelm
Sep 23, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Erik HanffDimitrios Tsikas
Jul 20, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·S KageN Ikeda
Apr 7, 2005·Journal of Inorganic Biochemistry·Benjamin P LuchsingerDavid J Singel
Aug 15, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Wenjuan S JobgenGuoyao Wu
Aug 12, 1999·Nitric Oxide : Biology and Chemistry·K SchulzM Kelm
Oct 16, 1999·Biochimica Et Biophysica Acta·I B ZavodnikM Bryszewska
Dec 29, 2004·Free Radical Research·Daniela GiustariniRanieri Rossi
Sep 29, 2004·Free Radical Biology & Medicine·Gregory R J ThatcherVioleta Toader
Sep 4, 2003·The Journal of Biological Chemistry·Enika NagababuJoseph M Rifkind
Oct 25, 2017·Bulletin of Experimental Biology and Medicine·V Yu TitovA N Osipov
Mar 9, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Marijke GrauPetra Kleinbongard
Jun 29, 1999·Analytical Chemistry·D J Anderson

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