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Determination of Nitrate and Nitrite by High-Performance Liquid Chromatography in Human Plasma

Vera Jedlicková, Zoltán Paluch, Stefan Alusík

J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Nov 15;780(1):193-7.

PMID: 12383495

Abstract:

A new, accurate, fast and simple method has been implemented by which nitrite and nitrate ions, as stable forms of nitric oxide production were studied. A study of these two ions was carried out by a sensitive and accurate HPLC method with two detectors. The most important advantages of the reported method are: short time of analysis, minimal sample pre-treatment, long life of the analytical column and stable eluent solution. The photodiode array UV-Vis detector detected nitrite and nitrate ions at an absorbance of 212 nm. Much more sensitive electrochemical detection with a WE (glassy carbon) electrode was used for the detection of nitrite ions. An analytical chromatographic column was formed by a sorbent, containing strong base anion-exchange groups bound in Cl(-) form in the hydrophilic hydroxyethyl methacrylate matrix. The anions were analysed in human plasma without deproteinization using 0.02 M sodium perchlorate monohydrate as eluent solution at pH 3.9. At this pH organic substances do not affect the analysis. The retention times for nitrite and nitrate were 3.62 and 3.72 min (by electrochemical detection) and 4.44 min, respectively. The method was linear (r=0.9992, 0.9998, 0.996) within a 1-100 (nitrate), 1-20 micro mol/l (nitrite) concentration range.

Chemicals Related in the Paper:

Catalog Number Product Name Structure CAS Number Price
AP7791073 Sodium perchlorate monohydrate Sodium perchlorate monohydrate 7791-07-3 Price
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