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Mixed-mode chromatography in pharmaceutical and biopharmaceutical applications

Authors : Kelly Zhanga, Xiaodong Liub


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.


Mixed-mode chromatography; Pharmaceutics; Biopharmaceutics

Citation: Kelly Zhanga, Xiaodong Liub Mixed-mode chromatography in pharmaceutical and biopharmaceutical applications doi:10.1016/j.jpba.2016.05.007

Received: 1 February 2016 Revised: 4 May 2016 Accepted: 5 May 2016 Available online: 6 May 2016

Copyright:© 2016 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (


We thank Xuefei Sun of Thermo Fisher Scientific for helping edit some of the references, Geoffrey Yeh and Sam Yang of Genentech for reviewing the manuscript.

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