Shnizer Sergei, Kagan Tamara, Lanir Amos, Maor Irit, Reznick Abraham Z
Lumitest Ltd, Nesher, Israel.
Luminescence. 2003 Mar-Apr;18(2):90-6. doi: 10.1002/bio.699.
Detection of electronically excited species (EES) in body fluids may constitute an important diagnostic tool in various pathologies. Examples of such products are triplet excited carbonyls (TEC), which can be a source for photon emission in the 400-550 nm range. The aim of the present study was to determine the actual contribution of lipid and protein components (protein carbonyls) to photon emission generated by thermochemiluminescence (TCL) during the heating of biological fluids. In this study, a new TCL Photometer device, designed by Lumitest Ltd, Israel, was used. Samples were heated to a constant temperature of 80 +/- 0.5 degrees C for 280 s and photon emission was measured at several time points. In order to compare the results of TCL measurements to conventional methods of detecting lipid and protein oxidation, each examined sample was also heated in a waterbath at 80 degrees C for 10-280 s. Lipid and protein oxidation were subsequently measured using conventional methods. The TCL of four polyunsaturated fatty acids (PUFA) with three to six double bonds was measured. The elevation of the PUFA TCL amplitude correlated with the increase in the number of double bonds of PUFA. A correlation between the increase in TCL intensity and protein carbonyl generation in bovine serum albumin (BSA) was also observed. In the venous blood serum, our study showed that an increase of TCL intensity during heating reflected the cleavage of TEC of lipid origin. Our study suggests that biological molecules such as proteins, lipids and other molecules, which may become unstable during heating, are capable of generating EES. We demonstrated that a TCL curve can be used as a kinetic model for measuring oxidative processes, which reflects modifications of different molecules involved in the oxidative stress phenomena.
检测体液中的电子激发态物种（EES）可能成为各种病理学中的一种重要诊断工具。这类产物的例子是三重态激发羰基化合物（TEC），它可以是400 - 550 nm范围内光子发射的来源。本研究的目的是确定脂质和蛋白质成分（蛋白质羰基）对生物流体加热过程中热化学发光（TCL）产生的光子发射的实际贡献。在本研究中，使用了以色列Lumitest Ltd公司设计的一种新型TCL光度计装置。将样品加热到80±0.5℃的恒定温度持续280秒，并在几个时间点测量光子发射。为了将TCL测量结果与检测脂质和蛋白质氧化的传统方法进行比较，每个检测样品还在80℃的水浴中加热10 - 280秒。随后使用传统方法测量脂质和蛋白质氧化。测量了具有三到六个双键的四种多不饱和脂肪酸（PUFA）的TCL。PUFA的TCL振幅升高与PUFA双键数量的增加相关。还观察到牛血清白蛋白（BSA）中TCL强度的增加与蛋白质羰基生成之间的相关性。在静脉血清中，我们的研究表明加热过程中TCL强度的增加反映了脂质来源的TEC的裂解。我们的研究表明，诸如蛋白质、脂质和其他分子等生物分子在加热过程中可能变得不稳定，它们能够产生EES。我们证明了TCL曲线可以用作测量氧化过程的动力学模型，该模型反映了参与氧化应激现象的不同分子的修饰。
