Analysis of specific lipid hydroperoxides by HPLC

Free fatty acid hydroperoxides and phospholipid hydroperoxides partition into the aqueous methanol layers, whereas triglyceride hydroperoxides and cholesterol ester hydroperoxides are recovered in 

9 ml of hexane supernatant layer evaporate dissolve in 450 p methanol/r-butan-ol (1 1, v/v) 

5 ml aqueous methanol layer filter through a 2 /im filter 

Inject 100 /A portions on RP18 column, elute with methanol/(-butanol (1 1 v/v). Detect hydro-peroxides etc. at 210 nm. 

Inject 50 /A on an aminophase column, elute wih methanol/40 mM sodium phosphate buffer pH 7.5,95 5 or 90 10 (v/v). Detect hydroperoxides at 265 nm. 

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