Wallin B, Camejo G
Astra Hässle AB, Mölndal, Sweden.
Scand J Clin Lab Invest. 1994 Jul;54(4):341-6. doi: 10.3109/00365519409087532.
The oxidative modification of lipoproteins is of clinical importance because of potential contribution to atherogenesis [1, 2, 3]. An early step in the complex process of oxidation is the peroxidation of polyunsaturated fatty acids. We describe here a method for the Cu(II)-catalyzed oxidation of human low density lipoproteins with the subsequent analysis of hydroperoxides formation in a single microtitre plate. The procedure includes a modification of an iodometric peroxide assay for test tubes using a commercially available reagent. The microtitre plate method correlated well with the test tube procedure (r = 0.99) and showed comparable sensitivity and reproducibility. It was sensitive down to 0.5 nmol hydroperoxides/well and linear up to at least 20 nmol well-1. The method can handle several hundreds of samples a day with considerably less labour than the test tube procedure. It was well suited to monitor the kinetics of lipoprotein oxidation. The method was also used to test the potency of antioxidants, however, some antioxidants may interfere with the iodometric reaction and should be tested in the assay before use.
脂蛋白的氧化修饰具有临床重要性，因为它可能对动脉粥样硬化的发生有促进作用[1,2,3]。氧化复杂过程的早期步骤是多不饱和脂肪酸的过氧化。我们在此描述一种用Cu(II)催化人低密度脂蛋白氧化的方法，随后在单个微量滴定板中分析氢过氧化物的形成。该程序包括使用市售试剂对用于试管的碘量法过氧化物测定法进行改进。微量滴定板法与试管法相关性良好(r = 0.99)，并显示出相当的灵敏度和重现性。它对低至0.5 nmol氢过氧化物/孔敏感，线性范围至少可达20 nmol/孔。该方法每天能处理数百个样品，比试管法省力得多。它非常适合监测脂蛋白氧化的动力学。该方法也用于测试抗氧化剂的效力，然而，一些抗氧化剂可能会干扰碘量反应，使用前应在测定中进行测试。
