Simultaneous determination of nitrite and nitrate anions in plasma, urine and cell culture supernatants by high-performance liquid chromatography with post-column reactions

Journal of Chromatography. B, Biomedical Applications
M N Muscará, G de Nucci

Abstract

A high-performance liquid chromatographic method for the determination of nitrite and nitrate anions derived from nitric oxide in biological fluids is presented. After separation on a strong anion-exchange column (Spherisorb SAX, 250 x 4.6 mm I.D., 5 microns), two on-line post-column reactions occur. The first involves nitrate reduction to nitrite on a copper-plated cadmium-filled column. In the second, the diazotization-coupling reaction between nitrite and the Griess reagent (0.05% naphtylethylendiamine dihydrochloride plus 0.5% sulphanilamide in 5% phosphoric acid) takes place, and the absorbance of the chromophore is read at 540 nm. This methodology was applied to biological fluids. Before injection into the chromatographic system, the samples were diluted and submitted to suitable clean-up procedures (urine and cell culture supernatant samples are passed through C18 cartridges, and serum samples were deproteinized by ultrafiltration through membranes with a molecular mass cut-off of 3000). The method has a sensitivity of 30 pmol for both anions, as little as 0.05-0.1 ml sample volume is required and linearity is observed up to 60 nmol for each anion.

References

Jun 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·P KubesD N Granger
Feb 1, 1990·Analytical Biochemistry·M J DunphyD J Smith
Jul 1, 1990·Food Additives and Contaminants·M J DennisR C Massey
Aug 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·D SalveminiJ R Vane
Jan 1, 1985·Die Nahrung·F Vlácil, I Vins
May 14, 1982·Journal of Chromatography·J P WitterE Balish
Oct 1, 1982·Analytical Biochemistry·L C GreenS R Tannenbaum

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Citations

Jul 1, 1998·Clinical Biochemistry·G EllisS K Makela
Mar 13, 1999·Clinical Biochemistry·P N BoriesL Dziedzic
Dec 19, 1996·European Journal of Pharmacology·A ZappelliniG De Nucci
Mar 26, 1997·European Journal of Pharmacology·M S FariaG De Nucci
Apr 25, 2000·Toxicon : Official Journal of the International Society on Toxinology·M L GuzzoS B Mello
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
Feb 22, 2000·British Journal of Pharmacology·M N MuscaráJ L Wallace
Aug 11, 2001·British Journal of Pharmacology·M N MuscaráJ L Wallace
Feb 1, 2006·Clinical and Experimental Pharmacology & Physiology·Shang Jyh KaoHsing I Chen
Jan 5, 2007·Clinical and Experimental Pharmacology & Physiology·Demeral D LiuHsing I Chen
Feb 28, 2007·Clinical and Experimental Pharmacology & Physiology·Chain Fa SuHsing I Chen
Apr 19, 2007·Clinical and Experimental Pharmacology & Physiology·Shang-Jyh KaoHsing I Chen
Jul 13, 2007·Biomedical Research·Nobuaki HanaueGen-yuki Yamane
Sep 5, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Dimitrios Tsikas
May 23, 2006·Pulmonary Pharmacology & Therapeutics·Chia-Hsiang ChuHsing I Chen
Jan 3, 2012·Immunopharmacology and Immunotoxicology·Luciana VismariJoão Palermo-Neto
Oct 31, 2006·Pulmonary Pharmacology & Therapeutics·I Chun ChuangHsing I Chen
Nov 5, 2008·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Hirotatsu Kojima
Sep 21, 2007·Clinical and Experimental Pharmacology & Physiology·Yen Chin LiuHsing I Chen
Feb 28, 2008·The Journal of Trauma·Shang-Jyh Kao, Hsing I Chen
Sep 20, 2007·Journal of Cardiovascular Pharmacology·Shang-Jyh KaoHsing I Chen
Mar 1, 2006·Critical Care Medicine·Hsing I ChenShang-Jyh Kao
Jan 11, 2001·Current Opinion in Clinical Nutrition and Metabolic Care·P ThérondA Legrand
Feb 9, 2000·American Journal of Physiology. Gastrointestinal and Liver Physiology·S L Freeman, W K MacNaughton
Jul 18, 2000·American Journal of Physiology. Gastrointestinal and Liver Physiology·D M McCaffertyP Kubes
Apr 6, 2000·American Journal of Physiology. Lung Cellular and Molecular Physiology·C TsujiH Nakazawa
Aug 3, 2000·American Journal of Physiology. Heart and Circulatory Physiology·M N MuscaráJ L Wallace
Jun 29, 1999·Analytical Chemistry·D J Anderson

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