High resolution 31P NMR of extracted phospholipids.

High resolution 31P NMR of extracted phospholipids.

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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 oolution light up of crude tissue lipid extracted from tissue sources using chloroform-methanol 2:1, the 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 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 cnd shellac spring 2023 routine analysis of phospholipids from biological sources.