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Journal of Lipid Research, Vol. 3, 378-381, July 1962
Copyright © 1962 by Lipid Research, Inc.

A sensitive and specific method for plasmalogens and other enol ethers

J. N. Williams Jr. , Carl E. Anderson , and Alice D. Jasik

Laboratory of Nutrition and Endocrinology, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Maryland and The Department of Biochemistry and Nutrition, School of Medicine, University of North Carolina, Ch

A spectrophotometric procedure for estimating plasmalogens and other enol ethers, based upon the specific reaction of the enol ether group with iodine, is presented. Optimum conditions for iodination with respect to methanol and KI concentration, pH, and time of incubation have been defined. After evaporating the solvent from the lipid extract containing 0.02 to 0.125 µmole plasmalogen, 0.9 ml methanol, 3.2 ml 0.094 m Na citrate (pH 5.5), 0.4 ml 3 m KI, and 0.5 ml 0.0005 m I₂ in 3 m KI are added and mixed. After standing for 40 min at room temperature, the mixture is extracted with 5 ml n-butyl acetate. Absorbancy of the butyl acetate layer is read at 363 mµ. Lipid and reagent blanks and an I₂ standard containing 0.25 µmole I₂ are treated similarly except that 0.9 ml 3 m KI is added to both blanks in place of I₂. Reproducibility is ±3.6% with 0.1 µmole levels of plasmalogen in lipid extracts. The range of enol ether estimated by this procedure is 0.02-0.125 µmole.

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