Fluorimetric determination of peroxynitrite based on an enzymatic reaction.
A novel fluorimetric method for the determination of peroxynitrite (ONOO-) using hemoglobin (Hb) as a catalyst is described. The method employs the reaction of ONOO with thiamine (TM), a colorless, non-fluorescent reagent in a glycine-NaCl-NaOH buffer solution (pH 12.7), to generate a highly fluorescent product, thiochrome (TC). The fluorescent product was monitored by fluorimetry. A linear calibration graph was obtained over an ONOO- concentration range from 4.95 x 10(-7) mol L(-1) to 2.97 x 10(-5) mol L(-1), with a detection limit of 9.78 x 10(-9) mol L(-1) ONOO-. The relative standard deviation at an ONOO- concentration of 2.11 x 10(-6) mol L(-1) was 4.15% (n = 9).
Cao
Qi Hua
QH
Department of Chemistry, Wuhan University, Wuhan 430072, People's Republic of China.
Zhou
Qian Xiong
QX
Cai
Ru Xiu
RX
Liu
Zhi Hong
ZH
D002021:Buffers; D002138:Calibration; D002384:Catalysis; D006454:Hemoglobins / Q000737:chemistry; D007202:Indicators and Reagents; D007700:Kinetics; D030421:Peroxynitrous Acid / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013831:Thiamine / Q000031:analogs & derivatives / Q000737:chemistry
D002021:Buffers; D006454:Hemoglobins; D007202:Indicators and Reagents; D030421:Peroxynitrous Acid; C005793:thiochrome; D013831:Thiamine