A sensitive, radiometric assay for lysophosphatidylcholine.
To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [3H]acetic anhydride and the [3H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [14C]lysophosphatidylcholine as an internal standard.
Dobmeyer
D J
DJ
Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Corr
P B
PB
Creer
M H
MH
D000089:Acetic Anhydrides; D000107:Acetylation; D000818:Animals; D002308:Cardiolipins; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003327:Coronary Disease / Q000378:metabolism; D004285:Dogs; D005227:Fatty Acids; D008244:Lysophosphatidylcholines / Q000032:analysis*; D008722:Methods; D009206:Myocardium / Q000166:cytology; D010955:Plasmalogens / Q000032:analysis*; D015203:Reproducibility of Results; D012588:Scintillation Counting; D013109:Sphingomyelins
D000089:Acetic Anhydrides; D002308:Cardiolipins; D005227:Fatty Acids; D008244:Lysophosphatidylcholines; D010955:Plasmalogens; D013109:Sphingomyelins; C032459:choline plasmalogens; C031800:acetic anhydride