Bottu G
Laboratorium voor Biochemie, Instituut voor Moleculaire Biologie, Vrije Universiteit Brussel, Belgium.
J Biolumin Chemilumin. 1991 Jul-Sep;6(3):147-51. doi: 10.1002/bio.1170060303.
The chemiluminescence of the system luminol +Fe2+ +H2O2 was measured in aqueous buffer at pH 7.2. In veronal (5,5-diethybarbiturate) buffer, the luminescence is strongly quenched by ethanol and mannitol, but only weakly by t-butanol, benzoate and superoxide dismutase (SOD); complexing Fe2+ with 1,20-phenanthroline or 2,2'-dipyridyl causes a decrease of light production that can be partially obviated by the simultaneous addition of SOD. In phosphate buffer, the luminescence is higher than in veronal and it is efficiently quenched by all four OH. quenchers and by SOD. In Tris buffer, no light production is observed as long as the Fe2+ is not complexed. When Fe2+ is complexed by pyrophosphate or phytate, there is a strong chemiluminescence in all three buffers, which is quenched by all four OH. quenchers and by SOD. When Fe2+ is complexed by EDTA or DTPA, very little luminescence is observed. The luminol analogue phthalhydrazide, which was suggested by Merényi and Lind as a reliable OH. detector, can replace luminol only in phosphate buffer, and thus turns out to be very specific indeed for free OH..
在pH 7.2的水性缓冲液中测量了鲁米诺+Fe²⁺+H₂O₂体系的化学发光。在巴比妥缓冲液（5,5 - 二乙基巴比妥酸盐）中，乙醇和甘露醇会强烈淬灭发光，但叔丁醇、苯甲酸盐和超氧化物歧化酶（SOD）的淬灭作用较弱；将Fe²⁺与1,10 - 菲咯啉或2,2'-联吡啶络合会导致发光量减少，同时添加SOD可部分消除这种减少。在磷酸盐缓冲液中，发光强度高于巴比妥缓冲液，并且会被所有四种羟基淬灭剂和SOD有效淬灭。在Tris缓冲液中，只要Fe²⁺未络合就观察不到发光。当Fe²⁺被焦磷酸盐或植酸盐络合时，在所有三种缓冲液中都会产生强烈的化学发光，并且会被所有四种羟基淬灭剂和SOD淬灭。当Fe²⁺被EDTA或DTPA络合时，观察到的发光很少。Merényi和Lind提出的鲁米诺类似物邻苯二甲酰肼作为一种可靠的羟基探测器，仅在磷酸盐缓冲液中可以替代鲁米诺，因此实际上对游离羟基非常特异。
