Analysis of solutions containing glutathione and inorganic nitrite: Application to nitroglycerin metabolism studies.

Original article


Curry SH, Aburawi SM, Whelpton R.

Division of Clinical Pharmacokinetics, College of Pharmacy, J. Hillis Miller Health Center, University of Florida, Gainesville, FL 32610, USA.

J Pharm Biomed Anal. 1987;5(3):275-82.


Nitroglycerin (GTN) is metabolized to 1,2-dinitroglycerin (1,2-GDN) and 1,3-dinitroglycerin (1,3-GDN) in vivo and in liver homogenates. 1,2-GDN and 1,3-GDN are converted to isomers of glyceryl mononitrate (GMN) in vivo. The denitration reactions yield inorganic nitrite (NO(-)(2)) which is oxidized to inorganic nitrate (NO(-)(3)). Denitration involves utilization of glutathione (GSH). In attempting to use the Bratton-Marshall assay for NO(-)(2) in studies of GTN metabolism in vitro, and in attempting to use Ellman’s reagent for GSH in the same research, apparent concentrations of both NO(-)(2) and GSH were noticed lower than anticipated. Apparent mutual interference by NO(-)(2) and GSH in their respective assays was then found. Development of a specific liquid chromatographic method for measurement of NO(-)(2), NO(-)(3), GSH and oxidized glutathione (GSSG) permitted the study of the interaction of NO(-)(2) and GSH, which yielded NO(-)(3) and GSSG.

Keywords: Nitroglycerin; glutathione; nitrate; nitrite; Bratton—Marshall assay; Ellman’s reagent; high-performance liquid chromatography