Miyazawa T, Saeki R, Inaba H
Department of Food Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.
J Biolumin Chemilumin. 1989 Jul;4(1):475-8. doi: 10.1002/bio.1170040162.
A combined system of chemiluminescence detection and high performance liquid chromatography (CL-HPLC) was developed to determine primary peroxidation products in biological tissues, such as phosphatidylcholine hydroperoxide (PCOOH). The CL-HPLC assay consists of separation of lipid classes with HPLC and detection of hydroperoxide-specific chemiluminescence. Hydroperoxides react with heme compounds to produce oxidants as suggested by our early studies on tissue low-level chemiluminescence in which singlet molecular oxygen is generated as one of the excited species in several biological systems involving free radical events. In the CL-HPLC method, a cytochrome c-luminol mixture was used as a hydroperoxide-specific luminescent reagent, and the quantification of hydroperoxide was performed by detecting chemiluminescence due to the luminol oxidation caused by the oxidant produced during the lipid hydroperoxides with heme. The detection limit of PCOOH was 10 pmole hydroperoxide-O2. PCOOH in normal human blood was found to be 10-500 pmol/ml plasma and significantly higher levels of PCOOH were observed in some hospitalized patients.
开发了一种化学发光检测与高效液相色谱联用系统（CL-HPLC），用于测定生物组织中的初级过氧化产物，如磷脂酰胆碱过氧化氢（PCOOH）。CL-HPLC分析包括用HPLC分离脂质类别以及检测过氧化氢特异性化学发光。如我们早期关于组织低水平化学发光的研究所表明的，过氧化氢与血红素化合物反应生成氧化剂，在涉及自由基事件的几个生物系统中，单线态分子氧作为激发态物种之一产生。在CL-HPLC方法中，细胞色素c-鲁米诺混合物用作过氧化氢特异性发光试剂，通过检测脂质过氧化氢与血红素产生的氧化剂引起的鲁米诺氧化导致的化学发光来进行过氧化氢的定量。PCOOH的检测限为10皮摩尔过氧化氢-O2。发现正常人血液中的PCOOH为10-500皮摩尔/毫升血浆，并且在一些住院患者中观察到PCOOH水平显著更高。
