We developed an isocratic reversed-phase HPLC method to measure arachidonic, palmitoleic, linoleic, eicosatrienoic, oleic, palmitic, and stearic acids from hydrolyzed erythrocytes. Washed erythrocytes were heated in methanol:HCl and the fatty acids extracted into hexane:amyl alcohol. After derivatization with 4-bromomethyl-7-methoxycoumarin, samples diluted in mobile phase (acetonitrile:water, 85:15 by vol) were injected onto a 250 x 4.6 mm C18 column, and the eluted fatty acids were detected fluorometrically. For all analytes, the mean within-batch CV was 8.2% (5.5-10.8%), the mean limit of detection was 7.0 mumol/L, a linear response was maintained up to 400 mumol/L, and results agreed well with those by gas chromatography. The addition of antioxidant (butylated hydroxytoluene) was essential for sample stability. We discuss hydrolysis and extraction times, derivatization temperature, critical steps in chromatography, and concentration units.
Despite IV linoleic acid administration, patients on long-term home parenteral nutrition have low erythrocyte stores of this essential fatty acid. This appears to be related to their low body fat stores. We suggest that they may be using much of the infused linoleic acid as an energy source and therefore are at risk of subclinical essential fatty acid deficiency.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.