Assessment of phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin in human serum.
We describe a method for analyzing the choline-containing phospholipids, namely phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin that is highly sensitive, easy to perform, and very reliable. The method employs enzymic hydrolysis of choline from these phospholipids by phospholipase D with subsequent oxidation of choline by choline oxidase and generation of hydrogen peroxide. Hydrogen peroxide is then reduced by peroxidase with p-hydroxy-phenylacetic acid acting as an oxygen acceptor to form a fluorescent product which is stable for at least 24 h without loss of linearity. The analysis requires a 5 microL sample volume and demonstrates linearity over a wide range (133 mumol/L-6.65 mmol/L). The CVs are less than 3%.
Fleming
J K
JK
College of Health Related Professions, Medical University of South Carolina, Charleston 29425.
Gadsden
R H
RH
D000293:Adolescent; D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008244:Lysophosphatidylcholines / Q000097:blood*; D008297:Male; D008875:Middle Aged; D010713:Phosphatidylcholines / Q000097:blood*; D013050:Spectrometry, Fluorescence; D013109:Sphingomyelins / Q000097:blood*; D013379:Substrate Specificity; D013696:Temperature
D007202:Indicators and Reagents; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D013109:Sphingomyelins