Dipalmitoylphoshatidylcholine-palmitic acid phase diagram studied by 13C nuclear magnetic resonance

Abstract

The phase diagram of dipalmitoylphosphatidylcholine (DPPC) and palmitic acid mixtures in excess D 2O was studied by 13C-NMR. Phase boundaries were determined from plots of apparent spin-spin relaxation time T 2 ∗ (for both choline methyl and fatty acid chain carbons) versus temperature. The peritectic transition in the 1–10 mol% region, whose existence has been theoretically inferred from the Gibbs phase rule but which was undetectable by differential thermal analysis (DTA) (S.E. Schullery et al. Biochemistry, 20 (1981) 6818–6824) was located by NMR at 41.6°C. A second, nearby peritectic line at 44°C, which had been shown by DTA to extend from about 3–25 mol% palmitic acid, was seen by NMR only above 10 mol%. The palmitic acid/DPPC complex (2:1), with a sharp melting point at 64°C, reported in earlier studies, was also seen by NMR. A phase diagram including both NMR and DTA results is presented. Important general conclusions from this study are: (i) NMR and scanning thermal analysis are complementary techniques for phase studies; each can see transitions that are invisible to the other. (ii) The case for the applicability of the Gibbs phase rule to lipid bilayer systems has been strengthened by the observance of two predicted, close-spaced boundaries. (iii) Low concentrations of fatty acids and related molecules can not be assumed to disperse as simple ideal solutes in the bilayer matrix.