48] Preparation and analysis of radiolabeled phosphatidylcholine and phosphatidylethanolamine containing 3H- and 14C-Labeled polyunsaturated fatty acids

Abstract

Publisher Summary This chapter discusses the preparation and analysis of radiolabeled phosphatidylcholine and phosphatidylethanolamine containing 3 H- and 14 C-labeled polyunsaturated fattyacids. Phosphatidylcholines with labeled fatty acids at either the 1 or the 2 position can be synthesized either by chemical acylation of monoacylglycerophosphocholine with fatty acid anhydrides or acyl chlorides or by enzymic acylation with fatty acyl-CoAs. The chemical methods have the advantage of yielding those with defined fatty acids, but involve synthesis on a larger scale and thus, require more labeled fatty acids. Furthermore, the yields in chemical syntheses of phosphatidylethanolamines with defined fatty acids have been relatively low in our hand. The enzymic method is suitable for biochemistry laboratories, but the products are contaminated with endogenous phosphatidylcholine or phosphatidylethanolamine from the enzyme preparation. This drawback of the latter method is minimized by using deoxycholate-treated microsomes as the enzyme source.