Peroxyoxalate chemiluminescence detection of condensates of malondialdehyde with thiobarbituric acids using a flow system.
The peroxyoxalate chemiluminescence(CL) detection method for the evaluation of the CL intensity of malondialdehyde(MDA) condensates with seven 2-thiobarbituric acid derivatives is described. The method consists of a flow injection technique together with a CL detection system using bis(2,4,6-trichlorophenyl) oxalate(TCPO) and hydrogen peroxide as chemiluminogenic reagents. Linear correlations between CL intensity and concentration are obtained for pmol levels of condensates. Among the condensates, 1,3-diethyl-2-thiobarbituric acid(DETBA)-MDA shows the largest CL intensity. High performance liquid chromatography (HPLC)/CL detection of DETBA-MDA and 1,3-diphenyl-2-thiobarbituric acid(DPTBA)-MDA using a mixture of TCPO and hydrogen peroxide in acetonitrile as a postcolumn reagent solution is also described. The detection limits for DETBA-MDA and DPTBA-MDA are 20 and 200 fmol, respectively, per 20 microL injection at a signal-to-noise ratio of 2. This HPLC/CL detection system was applied to the determination of MDA in rat brains by using DETBA as a fluorescent derivatizing reagent.
Nakashima
K
K
School of Pharmaceutical Sciences, Nagasaki University, Japan.
Nagata
M
M
Takahashi
M
M
Akiyama
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D017022:Flow Injection Analysis; D008163:Luminescent Measurements; D008315:Malondialdehyde / Q000032:analysis*; D010070:Oxalates / Q000737:chemistry*; D051381:Rats; D013858:Thiobarbiturates / Q000737:chemistry*
D010070:Oxalates; D013858:Thiobarbiturates; C048554:1,3-diphenyl-2-thiobarbituric acid; D008315:Malondialdehyde; C044622:peroxyoxalate; C046058:1,3-diethyl-2-thiobarbituric acid; C029684:thiobarbituric acid