An improved HPLC assay for phosphatidylcholine hydroperoxides (PCOOH) in human plasma with synthetic PCOOH as internal standard.
Quantitative and qualitative analyses of 1-palmitoyl-2-linoleoyl-phosphatidylcholine monohydroperoxide [PC 16:0/18:2-OOH] and 1-stearoyl-2-linoleoyl-phosphatidylcholine monohydroperoxide [PC 18:0/18:2-OOH] in human plasma were improved by chemiluminescence HPLC using synthetic 1-stearoyl-2-erucoyl-phosphatidylcholine monohydroperoxide (PC 18:0/22:1-OOH) as internal standard. The calibration curves of synthetic PC 16:0/18:2-OOH and PC 18:0/18:2-OOH, obtained by their direct injections with the IS into the HPLC system, were linear throughout the calibration range (10-1000 pmol). Within-day and between-day coefficients of variation were below 8%, and the recoveries were between 84% and 101%. Plasma concentrations of PC 16:0/18:2-OOH and PC 18:0/18:2-OOH were 102+/-59 nM (mean+/-SD) and 36+/-20 nM, respectively, in the 33 healthy volunteers. The present method might help understanding incompletely understood pathway of plasma phosphatidylcholine hydroperoxides.
Hui
Shu-Ping
SP
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan. keino@hoku-iryo-u.ac.jp
Chiba
Hitoshi
H
Sakurai
Toshihiro
T
Asakawa
Chitose
C
Nagasaka
Hironori
H
Murai
Tsuyoshi
T
Ide
Hajime
H
Kurosawa
Takao
T
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008054:Lipid Peroxides / Q000097:blood*; D008163:Luminescent Measurements / Q000379:methods; D008297:Male; D010713:Phosphatidylcholines / Q000097:blood* / Q000737:chemistry*; D010743:Phospholipids / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008054:Lipid Peroxides; D010713:Phosphatidylcholines; D010743:Phospholipids; C057590:phosphatidylcholine hydroperoxide