Analysis of cardiolipin molecular species by high-performance liquid chromatography of its derivative 1,3-bisphosphatidyl-2-benzoyl-sn-glycerol dimethyl ester.
Cardiolipin (CL, 1,3-bisphosphatidyl-sn-glycerol) is a four-acyl-chain phospholipid whose molecular species composition cannot be analyzed by standard procedures. Here we report a method to resolve the molecular species of CL by high-performance liquid chromatography of its derivative 1,3-bisphosphatidyl-2-benzoyl-sn-glycerol dimethyl ester. The CL derivative was characterized by 1H nuclear magnetic resonance spectroscopy, ultraviolet (uv) spectroscopy, thin-layer chromatography, and fatty acid analysis. The derivatization procedure did not change the fatty acid profile and provided a virtually complete conversion to the highly apolar, uv-visible product. In HPLC separations, recorded by 228 nm absorbance, a linear correlation was found between the area of individual peaks and their amount of lipid phosphorus. Bovine heart CL was resolved into 11 molecular species of which 6 (together accounting for 97 mol%) could be identified. The molecular species of bovine heart CL feature a linear relationship between their logarithmic retention time and their double bond number.
Schlame
M
M
Institute of Physical Biochemistry, University of Munich, Germany.
Otten
D
D
D002308:Cardiolipins / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D009682:Magnetic Resonance Spectroscopy; D010743:Phospholipids / Q000032:analysis
D002308:Cardiolipins; D010743:Phospholipids