Separation and quantitation of phospholipids in animal tissues by Iatroscan TLC/FID.
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 non-polar solvent mixture followed by partial scanning. Next, the rods are impregnated with oxalic acid, developed twice in CHCl3/CH3OH/CH3COOH/HCOOH/H2O (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 CHCl3/CH3OH/30% NH4OH (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.
De Schrijver
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Catholic University of Leuven, Laboratory of Nutrition, Belgium.
Vermeulen
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D000818:Animals; D002308:Cardiolipins / Q000302:isolation & purification; D002855:Chromatography, Thin Layer; D005410:Flame Ionization; D008244:Lysophosphatidylcholines / Q000302:isolation & purification; D010070:Oxalates; D019815:Oxalic Acid; D010712:Phosphatidic Acids / Q000302:isolation & purification; D010713:Phosphatidylcholines / Q000302:isolation & purification; D010714:Phosphatidylethanolamines / Q000302:isolation & purification; D010716:Phosphatidylinositols / Q000302:isolation & purification; D010718:Phosphatidylserines / Q000302:isolation & purification; D010743:Phospholipids / Q000302:isolation & purification*; D013109:Sphingomyelins / Q000302:isolation & purification
D002308:Cardiolipins; D008244:Lysophosphatidylcholines; D010070:Oxalates; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; D010743:Phospholipids; D013109:Sphingomyelins; D019815:Oxalic Acid