Mixed-mode chromatography in pharmaceutical and biopharmaceutical applications

Abstract

Mixed-mode chromatography (MMC) is a fast growing area in recent years, thanks to the new generation of mixed-mode stationary phases and better understanding of multimode interactions. MMC has superior applications in the separation of compounds that are not retained or not well resolved by typical reversed-phase LC methods, especially for polar and charged molecules. Due to the multiple retention modes that a single MMC column can offer, often MMC provides additional dimension to a separation method by adjusting the mobile phase conditions. Mixed-mode media is also an effective way to clean up complex sample matrices for purification purposes or for sensitive detection of trace amounts of analytes. In this article, we discuss mixed-mode stationary phases and separation mechanisms and review recent advances in pharmaceutical and biopharmaceutical applications including the analysis and/or purification of counterions, small molecule drugs, impurities, formulation excipients, peptides and proteins.

Styrene‐N‐phenylacrylamide co‐polymer modified silica monolith particles with an optimized mixing ratio of monomers as a new stationary phase for the separation of peptides in high performance liquid chromatography

“…Recently mixed‐mode chromatography (MMC) or multimode chromatography has shown to be a flexible and versatile approach for the simultaneous separation and analysis of variety of compounds . The driving force behind the development of mixed‐mode stationary phases is to achieve an alternative separation selectivity compared to conventional chromatographic modes such as reversed phase liquid chromatography.…”

“…The driving force behind the development of mixed‐mode stationary phases is to achieve an alternative separation selectivity compared to conventional chromatographic modes such as reversed phase liquid chromatography. MMC has superior applications in the separation of compounds that are not retained or not well resolved by typical reversed‐phase LC methods, especially for polar and charged molecules . Peptides are one of such analyte groups bearing importance in biological and pharmaceutical applications.…”

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

“…In mixed mode (multimodal) chromatography (MMC), compounds of interest in the mobile phase interact with the stationary phase materials through a combination of binding modes including ion exchange, hydrogen bonding, and hydrophobic interactions [142]. MMC methods have existed since the 1950s with hydroxyapatite chromatography combining CE and metal affinity [143].…”

“…MMC methods have existed since the 1950s with hydroxyapatite chromatography combining CE and metal affinity [143]. Although not a new idea, MMC is being increasingly developed for pharmaceutical/biopharmaceutical applications owing to the new generation of mixed-mode stationary phases and better understanding of multimodal interactions [142]. Most particlebased MMC media reported over recent years are based on silica gels [144] implemented for separation purposes.…”

Ion Pair Chromatograpy: A Critical Prespective

“…Today's chromatography market is flooded with different kinds of mixed mode columns [7] which are a convenient alternative to ion pair chromatography. For example polar embedded groups [8] have a positive or negative charge at the base of carbon chain which provide both ionic and reverse phase character and can possibly eliminate the need of an ion pair agent.…”