A simple and sensitive determination for plasmalogen lysophosphatidylethanolamine in rabbit platelets.
Plasmalogen lysophosphatidylethanolamine (LPE) in rabbit platelets was quantitatively determined as glycero-3-phosphorylethanolamine (GPE) after treatment with 5% trichloroacetic acid at 20 degrees C for two hours. GPE was measured directly using a high performance liquid chromatography with monitoring the fluorescence of o-phthaldialdehyde/2-mercaptoethanol adducts. The assay was sensitive to 50 pmol of plasmalogen LPE and was linear over a 200-fold concentration range. While, 1-acyl LPE and phosphatidylethanolamine from bovine brain (containing about 50% plasmalogen) were hardly cleaved to GPE under the same conditions. These procedures were specific for plasmalogen LPE, and simpler and more sensitive in comparison with conventional analytical methods, e.g., two-dimensional thin layer chromatography with subsequent phosphorus assay. Employing the present method, plasmalogen LPE was found to increase rapidly and to decrease subsequently when rabbit platelets were stimulated by thrombin.
Morikawa
S
S
Department of Biochemistry, Shiga University of Medical Science, Japan.
Taniguchi
S
S
Fujiwara
M
M
Kumada
K
K
Ishida
T
T
Horiike
K
K
Nozaki
M
M
D000818:Animals; D001792:Blood Platelets / Q000032:analysis* / Q000187:drug effects; D001923:Brain Chemistry; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D005456:Fluorescent Dyes; D008246:Lysophospholipids / Q000032:analysis*; D008563:Membrane Lipids / Q000032:analysis*; D010714:Phosphatidylethanolamines / Q000032:analysis / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism; D010955:Plasmalogens / Q000032:analysis*; D011817:Rabbits; D013917:Thrombin / Q000494:pharmacology
D005456:Fluorescent Dyes; D008246:Lysophospholipids; D008563:Membrane Lipids; D010714:Phosphatidylethanolamines; D010955:Plasmalogens; C008301:lysophosphatidylethanolamine; C002449:glycerophosphoethanolamine; D010741:Phospholipases A; D013917:Thrombin; D002118:Calcium