Olszowski S, Olszowska E, Stelmaszyńska T, Krawczyk A
Jagiellonian University College of Medicine, Institute of Medical Biochemistry, 31-034 Krakow, ul Kopernika 7, Poland.
Luminescence. 1999 May-Jun;14(3):139-45. doi: 10.1002/(SICI)1522-7243(199905/06)14:3<139::AID-BIO531>3.0.CO;2-A.
Light is emitted in systems containing N-chloramines or hypochlorite, H(2)O(2) and N-(4-aminobutyl)-N-ethylisoluminol (ABEI). The emission is enhanced by 4-iodophenol (PIP) in alkaline solution (1 mol/L NaOH), while at lower pH range (9-11) PIP is not only inactive but also its presence reduces chemiluminescence (CL) of the monochloramine-H(2)O(2)-ABEI system to the background. Two procedures for ABEI-labelled IgG assays were developed, with PIP in 1 mol/L NaOH and without PIP at pH 11, and the standard curves of free ABEI in these conditions were examined. We suggest also that the oxidative deamination of taurine chloramine leads to the formation of the various carbonyl derivatives and their formation is accelerated in the presence of H(2)O(2), especially in less alkaline solutions (pH 11). Moreover, the formation of enol forms of aldehydes in assay buffers was observed. The yield of the phenoxy radical mediators of ABEI oxidation and the pH-dependent H(2)O(2):HO(2) ratio seems to be decisive for the overall CL in the system examined. The main advantage of this method is that CL does not need precise timing of measurements and assays can be performed over a long period of time (hours) using a plate luminometer.
在含有N - 氯胺或次氯酸盐、H₂O₂和N -（4 - 氨基丁基）- N - 乙基异鲁米诺（ABEI）的体系中会发光。在碱性溶液（1mol/L NaOH）中，4 - 碘苯酚（PIP）可增强这种发光，而在较低pH范围（9 - 11）时，PIP不仅无活性，而且其存在会将一氯胺 - H₂O₂ - ABEI体系的化学发光（CL）降低至背景水平。开发了两种用于ABEI标记IgG测定的方法，一种是在1mol/L NaOH中加入PIP，另一种是在pH 11时不加入PIP，并检测了这些条件下游离ABEI的标准曲线。我们还提出，牛磺酸氯胺的氧化脱氨基作用会导致各种羰基衍生物的形成，并且在H₂O₂存在下，尤其是在碱性较弱的溶液（pH 11）中，其形成会加速。此外，在测定缓冲液中观察到了醛的烯醇形式的形成。ABEI氧化的苯氧基自由基介质的产率以及pH依赖性的H₂O₂:HO₂比值似乎对所研究体系中的整体CL起决定性作用。该方法的主要优点是CL测定不需要精确计时，并且可以使用平板发光计在很长一段时间（数小时）内进行测定。
