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Separation and quantitation of phospholipids in animal tissues by latroscan TLC/FID
Author(s) -
De Schrijver Remi,
Vermeulen Daniel
Publication year - 1991
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02544028
Subject(s) - phosphatidic acid , chemistry , phosphatidylethanolamine , chromatography , phospholipid , phosphatidylserine , phosphatidylcholine , phosphatidylinositol , sphingomyelin , calibration curve , lysophosphatidylcholine , analytical chemistry (journal) , quantitative analysis (chemistry) , oxalic acid , detection limit , biochemistry , membrane , kinase
A procedure has been developed to separate and quantitate phospholipids, including phosphatidylinositol and phosphatidylserine, from animal tissues by means of the Iatroscan TLC/FID technique. The method is based on the use of 0.01 M oxalic acid impregnated Chromarods‐SII and stepwise resolution of the phospholipids in the presence of 1,2‐dipalmitoyl‐ sn ‐glycero‐3‐phospho ( N,N ‐dimethylethanolamine) as internal standard. To remove the neutral lipids, the rods are initially developed in a nonpolar solvent mixture followed by partial scanning. Next, the rods are impregnated with oxalic acid, developed twice in CHCl 3 /CH 3 OH/CH 3 COOH/HCOOH/H 2 O (80∶35∶2∶1∶3, v/v/v/v/v) and partially scanned for measuring lysophosphatidylcholine, sphingomyelin and phosphatidylcholine. The subsequent step involves double development in CHCl 3 /CH 3 OH/30% NH 4 OH (60∶35∶0.9, v/v/v) to resolve cardiolipin, internal standard, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and phosphatidic acid. For each phospholipid a linear calibration curve with a highly significant correlation coefficient was obtained. However, the calibration lines extrapolated to negative intercepts on the ordinate, indicating declining sensitivity at low phospholipid loads.