New method for analyzing the nitrite level in PC12 cells using capillary electrophoresis.
Nitric oxide (NO) is a gaseous molecule shown to have signal transmitter properties in organisms. Direct measurement of NO in physiological conditions has been difficult due to its short lifetime and low concentration. Nitrite has been used as a marker for NO formation in biological systems. Capillary electrophoresis (CE) has been recently used to measure nitrite in biological fluids. The purpose of this study is to analyze nitrite in PC12 cells (pheochromocytoma cell line) using CE. Optimal CE performance was employed with 150 mM Tris-phosphate, 6 microM hexadecyltrimethyammonium chloride buffer at pH 7.0 and a fused-silica column of 57 cm x 75 microm I.D. The signal was measured with a UV detector at 214-nm wavelength and negative potential of 10 kV was applied for nitrite analysis. Under the optimum conditions, we monitored the changes in the concentration of the nitrite levels through synergistic stimulation of tumor necrosis factor alpha plus gamma-interferon in PC12 cells.
Lee
Jeeyeon
J
Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, PO Box 131, Cheongryang, Seoul 130-650, South Korea.
Ban
Eunmi
E
Yi
Seh-Yoon
SY
Yoo
Young Sook
YS
D000818:Animals; D002021:Buffers; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D009573:Nitrites / Q000032:analysis*; D016716:PC12 Cells; D051381:Rats; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D002021:Buffers; D002593:Cetrimonium Compounds; D009573:Nitrites; D000077286:Cetrimonium