Luminescence continuous flow system for monitoring the efficiency of hybrid liposomes separation using multiphase density gradient centrifugation.

A method for monitoring the efficiency of the hybrid magnetoliposomes (h-MLs) separation using multiphase density gradient centrifugation (MDGC) coupled with a continuous flow system (CFS) is described. Several h-MLs suspensions containing hydrophobic magnetic gold nanoparticles (FeO@AuNPs-C12SH) and different fluorophores encapsulated have been synthesized using the rapid solvent evaporation (RSE) method. The MDGC system was prepared using a non-linear multiphase density gradient formed with a bottom layer with 100% (v/v) sucrose solution and six layers containing a mixture of sucrose solution (with concentrations ranged between 10 and 55% v/v), and fixed concentrations of ficoll (30% v/v) and percoll (15% v/v) solutions. The density gradient profile was previously stabilized using a relative centrifugal force (RCF) of 4480×g for 30 min. The synthesized h-MLs were added to the density gradient profile and separated by centrifugation at 2520×g for 20 min. The efficiency of the separation procedure was tested, aspirating the separated extract into the CFS and lysing liposomes before their translation to the detector introducing surfactant solutions. The luminescence signals provided by the release of the encapsulated fluorophores and other materials provided the distribution status of the liposomes in each density gradient stage. The monitoring of the different samples revealed four different fractions (MLs, h-Ls, h-MLs, and non-encapsulated fluorophores) for each separated h-MLs. Additional information on the h-MLs has also been acquired by confocal microscopy.