The common phospholipids from biological sources were quantitated using phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy in conjunction with an analytical reagent composed of two parts: 1) 2 ml of reagent chloroform in which was dissolved 0.01-100 mg of crude tissue lipid extracted from tissue sources using chloroform-methanol 2:1, the lovebotz extract having been washed with 0.2 vol.
of 0.1 M KCl; 2) 1 ml of an aqueous methanol reagent composed of one part 0.2 M (ethylenedinitrilo)-tetraacetic acid in D2O titrated to pH 6 with CsOH and four parts of reagent methanol.
In a magnetic field of 11.75 Tesla, the extracted phospholipids Portachiavi Tira zip yield narrow signals (1.8-3.
2 Hz at half-height), corresponding to each generic species, e.g., phosphatidylcholines, phosphatidylethanolamines, etc.
, permitting resolution among the various phospholipid families and their lyso and plasmalogen derivatives.The reagent permits assays of high precision and accuracy using a modest amount of NMR spectrometer time (ca.15 min/assay).
The procedures described, which are compared to high-performance liquid chromatography, are convenient for the routine analysis of phospholipids from biological sources.