Detection of chemiluminescence in lipid peroxidation of biological systems and its application to HPLC

✍ Scribed by Miyazawa, Teruo ;Saeki, Rie ;Inaba, Humio

• Publisher: John Wiley and Sons
• Year: 1989
• Weight: 250 KB
• Volume: 4
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170040162

✦ Synopsis

A combined system of chemiluminescence detection and high performance liquid chromatography (CL-HPLC) was developed t o 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 oluminol mixture was used as a hydroperoxide-specific luminescent reagent, and the quantification of hydroperoxide was performed by detecting chemiluminescence due t o the luminol oxidation caused by the oxidant produced during the lipid hydroperoxides with heme. The detection limit of PCOOH was 10 pmole hydroperoxide-02. 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.