Determination of endothelium-derived nitrite/nitrate by gas chromatography/tandem mass spectrometry using (15N)NaNO2 as internal standard

✍ Scribed by F.-M. Gutzki; D. Tsikas; U. Alheid; J. C. Frölich

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 472 KB
• Volume: 21
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/bms.1200210207

✦ Synopsis

A sensitive gas chromatographic/tandem mass spectrometric method for the determination of nitrite and nitrate in endothelial cell column perfusates was developed. Nitritelnitrate were converted in the presence of the internal standard ("N)NaNO, to the corresponding l-nitro-2,4,Btrimethoxybenzene (NTMB) derivatives which were subsequently analysed by gas chromatography/tandem mass spectrometry, monitoring m/z = 46 and m/z = 47, respectively. The good reproducibility, the wide linear range of the calibration curve from 25 ng ml-' to 2500 ng ml-' nitrite and the low detection limit of 200 pg NTMB allowed a sensitive and accurate quantification of nitrite/nitrate in perfusates of endothelial cell columns. Bradykinin bad a stimulating effect on human and bovine endothelial cells and led to a threefold increase of nitritelnitrate in endothelial cell column perfusates compared to those of unstimulated cells.