PEGylated gold nanorod separation based on aspect ratio: characterization by asymmetric-flow field flow fractionation with UV-Vis detection

The development of highly efficient asymmetric-flow field flow fractionation (A4F) methodology for biocompatible PEGylated gold nanorods (GNR) without the need for surfactants in the mobile phase is presented. We report on the potential of A4F for rapid separation by evaluating the efficiency of functionalized surface coverage in terms of fractionation, retention time (tR) shifts, and population analysis. By optimizing the fractionation conditions, we observed that the mechanism of separation for PEGylated GNRs by A4F is the same as that for CTAB stabilized GNRs (i.e., according to their AR) which confirms that the elution mechanism is not dependent on the surface charge of the analytes and/or the membrane. In addition, we demonstrated that A4F can distinguish different surface coverage populations of PEGylated GNRs. The data established that a change in Mw of the functional group and/or surface orientation can be detected and fractionated by A4F. The findings in this study provide the foundation for a complete separation and physicochemical analysis of GNRs and their surface coatings, which can provide accurate and reproducible characterization critical to advancing biomedical research.