Reactions of nitrite in erythrocyte suspensions measured by membrane inlet mass spectrometry.
The reactions of nitrite with deoxygenated human erythrocytes were examined using membrane inlet mass spectrometry to detect the accumulation of NO in an extracellular solution. In this method an inlet utilizing a silicon rubber membrane is submerged in cell suspensions and allows NO to pass from the extracellular solution into the mass spectrometer. This provides a direct, continuous, and quantitative determination of nitric oxide concentrations over long periods without the necessity of purging the suspension with inert gas. We have not observed accumulation of NO compared with controls on a physiologically relevant time scale and conclude that, within the limitations of the mass spectrometric method and our experimental conditions, erythrocytes do not generate a net efflux of NO after the addition of millimolar concentrations of nitrite. Moreover, there was no evidence at the mass spectrometer of the accumulation of a peak at mass 76 that would indicate N(2)O(3), an intermediate that decays into NO and NO(2). Inhibition of red cell membrane anion exchangers and aquaporins did not affect these processes.
Mikulski
Rose
R
Department of Pharmacology and Therapeutics, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Tu
Chingkuang
C
Swenson
Erik R
ER
Silverman
David N
DN
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D001457:Anion Exchange Protein 1, Erythrocyte / Q000037:antagonists & inhibitors; D020346:Aquaporins / Q000037:antagonists & inhibitors; D001672:Biocompatible Materials; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D008567:Membranes, Artificial; D009569:Nitric Oxide / Q000378:metabolism; D011485:Protein Binding; D012977:Sodium Nitrite / Q000737:chemistry / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D020346:Aquaporins; D001672:Biocompatible Materials; D006454:Hemoglobins; D008567:Membranes, Artificial; D009569:Nitric Oxide; D002731:4-Chloromercuribenzenesulfonate; C100003:deoxyhemoglobin; D012977:Sodium Nitrite; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid