Related Experiment Video
Updated: May 19, 2026

Lipid Vesicle-mediated Affinity Chromatography using Magnetic Activated Cell Sorting (LIMACS): a Novel Method to Analyze Protein-lipid Interaction
Published on: April 26, 2011
Utilization of hydrophilic-interaction LC to minimize matrix effects caused by phospholipids
Beijing Tan1, Andre Negahban, Thomas McDonald
1Dynamics & Metabolism Department, Pfizer Global Research & Development, Groton, CT 06340, USA. beijing.tan@pfizer.com
Background:
In bioanalysis, phospholipids may affect the precision and accuracy of LC-MS/MS methods and compromise the quality of the results, especially when samples in complex biomatrices are extracted by protein precipitation techniques.
Results:
It was found that the retentive behavior of both common pharmaceuticals and physiologically relevant phospholipids under bare silica hydrophilic-interaction LC (HILIC) is more predictable than under reversed-phase conditions. In particular, the retention time of phospholipids was not significantly affected by varying the salt and acid modifiers in the mobile phases, but common pharmaceuticals can be shifted away from these phospholipid interferences through mobile phase modifiers. Several mass spectrometric techniques were applied to confirm this finding.
Conclusion:
HILIC chromatography is a valued tool in the development of robust bioanalytical assays with minimal and predictable phospholipid interferences. Furthermore, addition of a small amount of ion-pairing additives can reliably move pharmaceutical compounds away from these suppressive regions.
Related Concept Videos
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process
High-Performance Liquid Chromatography: Instrumentation
Thin-Layer Chromatography (TLC): Overview
To begin the analysis, a mixture of compounds is spotted on the starting line on the TLC plate using a thin capillary. The bottom of the...

