An Aromatic Phosphine Reagent for the HPLC-Fluorescence Determination of Hydroperoxides - Determination of Phosphatidylcholine Hydroperoxides in Human Plasma

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Abstract
A new method is developed to determine picomole levels of phosphatidylcholine hydroperoxides (PC-HPO) in human plasma. The PC-HPO was extracted from 500 μ1 of human plasma with a mixture of chloroform and methanol and separated on a HPLC with a silica gel column using the chloroform - methanol - water system. Detection was carried out by fluorometry after post column derivatization with diphenyl-1-pyrenylphosphine CDPPP). The determination of PC-HPO was performed by the method of standard addition. The sampling error was corrected by using the UV peak of inherent unoxidized PC as an internal standard. By this method, the PC-HPO levels in healthy adult human plasma were 20–40 pmol/ml which compares favorably to the normal expected values.