Enzymic assay for phosphatidylcholine and sphingomyelin in serum.
We describe a method for the direct enzymatic determination of phosphatidylcholine and sphingomyelin in serum. Phospholipase C (EC 3.1.4.3) and sphingomyelinase (EC 3.1.4.12) are used to hydrolyze phosphatidylcholine and sphingomyelin, respectively, with high specificity; alkaline phosphatase (EC 3.1.3.1) is used to cleave inorganic phosphorus. The choline group, after oxidation with choline oxidase (EC 1.1.3.17), is detected with 4-aminoantipyrine. Alternatively, phosphorus is assayed with metavanadate. The excellent agreement between results of this modification of a procedure described by Artiss et al. (Microchem. J. 26: 1017, 1980) for amniotic fluid and of the conventional thin-layer chromatographic method makes this an attractive method for determination of both phospholipid subclasses in serum.
D000429:Alcohol Oxidoreductases; D002855:Chromatography, Thin Layer; D006801:Humans; D008722:Methods; D010713:Phosphatidylcholines / Q000097:blood*; D013108:Sphingomyelin Phosphodiesterase; D013109:Sphingomyelins / Q000097:blood*; D010738:Type C Phospholipases
D010713:Phosphatidylcholines; D013109:Sphingomyelins; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D010738:Type C Phospholipases; D013108:Sphingomyelin Phosphodiesterase