Previous work in this laboratory using Cholesteryl ester vesicles to encapsulate and deliver a wide variety of substances (e.g. chromophores, antibiotics, peptides, IgG antibodies and plasmid DNA) has shown cholesteryl ester vesicle-mediated delivery of FITC-labelled peptides into various mouse tissues (including brain) after oral administration.  Vital to the analysis of these vesicles is the determination and quantitation of ester composition using High Performance Liquid Chromatography (HPLC). Ongoing work has resulted in development of methods for a limited number of cholesteryl esters, both short (C6-C10) and moderate (C12-C16) alkyl chain lengths. These methods were for chains of even number length.  The present research project expands the development of HPLC analyses to include esters with alkyl chains of odd number length, from C9 to C19.  The current lipid analysis method was the start point for quantitative analysis of the odd chain length esters.  Initial work has included modification of the mobile phase solvents to determine ideal solubility of the esters that will optimize quantitative results using the HPLC.  Current methods for the short and even chains appears sufficient for chain lengths C9 to C13 as determined by standard curve determination of known concentrations.  However, the longer chain esters have differential solubility characteristics as well as weaker detection at 205nm.  Determination of the optimum solubility for the longer chain esters is critical for quantitative HPLC results.  These methods showed linear standard curves, which will allow for determination of lipid content of cholesteryl ester vesicles with the longer odd number alkyl chain esters.

Development of HPLC methods for analysis of cholesteryl esters with alkyl chains of odd number length

256th National Meeting (2018)

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