New technique to quantify phospolipids in disordered phase in lung surfactant by electron spin resonance

In pulmonary surfactant (PS) the coexistence of the ordered (Lo) and disordered (Ld) lipid phases would be essential for an optimal activity. Electron spin resonance (ESR) of PS labeled with 5-doxil stearic acid shows two spectral components called S and W. S/W ratio could be understood as the ratio between the probe population in Lo and in Ld. Although the specificity of S/W as an indicator of Lo/Ld has not yet been demonstrated, S/W has been used qualitatively to study changes in Lo/Ld. The goal of this paper is to stablish the correlation between S/W parameter and the amount of lipids in Ld (%Ld) measured by the ESR TEMPO technique described in our previous work. S/W and %Ld were measured in different PS samples under different experimental conditions. The results showed an inverse correlation between S/W and %Ld (r = -0.983; p < 0.001). We demonstrated that the S/W is sensible to the changes of Lo/Ld and can be used to quantify the %Ld.