Quantitative determination of catecholamines in individual bovine adrenomedullary cells by reversed-phase microcolumn liquid chromatography with electrochemical detection.
A microcolumn liquid chromatographic method for the determination of norepinephrine and epinephrine in single bovine adrenomedullary cells is described. A single cell is isolated from a culture plate, (3,4-dihydroxybenzyl)amine is added as an internal standard, and the cell is lysed with perchloric acid. After centrifugation, the supernatant is injected directly into a 42 or 50 microns Inner diameter C18 reversed-phase column operated with electrochemical detection. Detection limits for norepinephrine and epinephrine were 46 and 75 amol, respectively. Of the 22 cells examined, 8 contained predominantly norepinephrine, 10 contained predominantly epinephrine, and at least 4 cells contained significant amounts of each. On average, a single cell contained approximately 150 fmol of catecholamine.
Cooper
B R
BR
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290.
Jankowski
J A
JA
Leszczyszyn
D J
DJ
Wightman
R M
RM
Jorgenson
J W
JW
D000313:Adrenal Medulla / Q000737:chemistry* / Q000166:cytology; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D004837:Epinephrine / Q000032:analysis*; D009638:Norepinephrine / Q000032:analysis*
D009638:Norepinephrine; D004837:Epinephrine