Liposome/saline partition coefficients of low-molecular-weight solutes by gel chromatography

A chromatographic method for the determination of association constants of rapidly dissociable complexes is described and applied to quantification of liposome/saline partition coefficients using gel chromatography. The approach allows for estimation of the free solute concentration in the sample by simple manual processing of the intact right-hand part of the solute peak deformed due to gradual diffusion of the accumulated solute from the liposomes along the separation column. Validity of the procedure was confirmed by both reasonable agreement with equilibrium dialysis data and model-based deconvolution of the distorted peak into its two components corresponding to initially unbound compound and to that escaped from the liposomes during the separation process.