Micellar HPLC: Investigation of the Retention of Positively Charged Peptides Using Cationic Micellar Mobile Phases

Abstract

Many new and complex molecules are being investigated as potential drug candidates. Conventional analytical methodologies may not be suitable for determining the purity, identity, and degradation of these complex molecules. Therefore, new analytical techniques must be developed that address and overcome these problems. Micellar liquid chromatography is such a technique.

Micellar liquid chromatography (MLC) is a technique where a micellar agent is added to a mobile phase that contains a buffer and a small amount of organic modifier. Several advantages are apparent with MLC when compared to reversed-phase liquid chromatography. MLC uses a much lower amount of organic modifier and is therefore less toxic, MLC does not denature peptides and proteins as does RPLC, and gradient MLC is done without the need for long column re-equilibration. In this study, various mobile phase variables were studied to determine the effect that each had on peptide retention.

The variables studied include: the concentration of micellar agent, mobile phase ionic strength, concentration of buffer, concentration of organic modifier and mobile phase pH. The results that were obtained are discussed.