Small-angle neutron scattering study of lipid phase diagrams by the contrast variation method

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Abstract
A small-angle neutron scattering study of slightly sonicated liposomes of binary lipid mixtures is presented. The neutron scattering of lipid lamellae may be analyzed in terms of the Kratky-Porod model of scattering by 2-dimensional systems. The contrast variation technique may thus be applied in order to study the structure and phase diagrams of lipid layers not disturbed by heavy sonication. The thickness of isolated bilayers is measured and molar volumes of pure lipid phases are determined. Mixtures of deuterated dimyristoylphosphatidylcholine with protonated dipalmitoylphosphatidylcholine and protonated distearoylphosphatidylcholine, respectively, are studied. Excess volumes of lipid mixtures, are determined by the contrast variation. For the 1st mixture, positive excess volumes of +86 .ANG.3 in the crystalline phase (5.degree. C) and of +49 .ANG.3 in the fluid phase (35.degree. C) are obtained. These large positive excess volumes are interpreted in terms of free volume creation at the interface between the monolayers, which indicates that the polar head groups are rather fixed with respect to the lipid-water interface. The phase boundaries at a given temperature may be determined by performing contrast variation experiments for 2 mixtures with different initial composition. Good agreement with existing experimental data is observed for the 1st mixture. A miscibility gap is established in the crystalline state of the 2nd mixture. A most interesting result is the finding of an immiscibility in the fluid state. This is interpreted in terms of critical concentration fluctuations caused by the critical demixing point of the solid-state miscibility gap hidden below the liquidus line.