Chokshi H P, Barbush M, Carlson R G, Givens R S, Kuwana T, Schowen R L
Center for Bioanalytical Research, University of Kansas, Lawrence 66045-0046.
Biomed Chromatogr. 1990 May;4(3):96-9. doi: 10.1002/bmc.1130040304.
The mechanism of the oxalate/hydrogen peroxide chemiluminescence reaction has been examined by magnetic resonance techniques. Investigation of the reactive intermediates involved in chemiluminescence was carried out with bis(2,6-difluorophenyl)oxalate (DFPO) using 19F NMR to probe its reactions with aqueous hydrogen peroxide. Formation and reactions of the intermediate hydroperoxy oxalate ester B along with the formation of the half ester product C and difluorophenol D were monitored by 19F NMR. When the reaction of DFPO and aqueous hydrogen peroxide was carried out in the presence of dansylphenylalanine, a typical fluorescent analyte, the intensity of the resonance due to the intermediate B was diminished in direct proportion to the concentration of the analyte. Comparison of the time/intensity profile of the chemiluminescence emission with that of the 19F NMR transient suggests that the hydroperoxy oxalate ester B is the likely 'reactive' intermediate, capable of participating in a chemically initiated electron exchange luminescence mechanism.
通过磁共振技术研究了草酸酯/过氧化氢化学发光反应的机理。使用19F核磁共振研究双(2,6-二氟苯基)草酸酯(DFPO)与过氧化氢水溶液反应中涉及的化学发光反应中间体。通过19F核磁共振监测中间体氢过氧草酸酯B的形成和反应，以及半酯产物C和二氟苯酚D的形成。当DFPO与过氧化氢水溶液的反应在典型荧光分析物丹磺酰苯丙氨酸存在下进行时，中间体B引起的共振强度与分析物浓度成正比降低。化学发光发射的时间/强度曲线与19F核磁共振瞬态曲线的比较表明，氢过氧草酸酯B可能是能够参与化学引发电子交换发光机制的“活性”中间体。
