Determination of myoglobin based on its enzymatic activity by stopped-flow spectrophotometry.
A new method has been developed for the determination of myoglobin (Mb) based on its enzymatic activity for the oxidation of o-phenylenediamine (OPDA) with hydrogen peroxide. Stopped-flow spectrophotometry was used to study the kinetic behavior of the oxidation reaction. The catalytic activity of Mb was compared to other three kinds of catalyst. The time dependent absorbance of the reaction product, 2,3-diamimophenazine (DAPN), at a wavelength of 426 nm was recorded. The initial reaction rate obtained at 40 degrees C was found to be proportional to the concentration of Mb in the range of 1.0 x 10(-6) to 4.0 x 10(-9)mol L(-1). The detection limit of Mb was found to be 9.93 x 10(-10)mol L(-1). The relative standard deviations were within 5% for the determination of different concentrations of Mb. Excess of bovine serum albumin (BSA), Ca(II), Mg(II), Cu(II), glucose, caffeine, lactose and uric acid did not interfere.
Zheng
Qi
Q
Department of Chemistry, Wuhan University, Wuhan 430072, PR China. zq.1101@sina.com
Liu
Zhihong
Z
Cai
Ruxiu
R
D000818:Animals; D002384:Catalysis; D002417:Cattle; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007700:Kinetics; D009211:Myoglobin / Q000032:analysis*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D010619:Phenazines / Q000032:analysis; D010655:Phenylenediamines / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods*; D014527:Uric Acid / Q000378:metabolism
D009211:Myoglobin; D016877:Oxidants; D010619:Phenazines; D010655:Phenylenediamines; C054312:2,3-diaminophenazine; D014527:Uric Acid; D012710:Serum Albumin, Bovine; C034193:1,2-diaminobenzene; D006861:Hydrogen Peroxide