Yuliya E. Silina
INM-Leibniz Institute for New Materials, Germany
Title: Hydrophilic interaction chromatography-tandem mass spectrometry for the cellular analysis: opportunities and challenges
Biography
Biography: Yuliya E. Silina
Abstract
Recently, a link between changes in the cellular state refl ected by nucleotide and lipid profi les has been established. Th e
analytical determination of nucleotides is not trivial, however, because of their high polarity and hydrophilic nature. Liquid
chromatography (LC) has been widely implemented for determination of nucleotides from biological samples. Most of these
LC-based techniques usually require ion-pairing reagents, thus making them unsuitable for LC-mass spectrometry (MS), timeconsuming,
however, and also exhibited limited chromatographic resolving and strong matrix background power for biological
samples. On the other hand, because of complexity of lipids, lipids analysis is still full of challenges. Meanwhile, due to the
vital roles of the changes in human physiological and pathological process, lipidomics is attracting more and more attention.
Hydrophilic interaction chromatography (HILIC) has become a powerful tool for the retention of polar analytes, because
of its excellent mobile phase compatibility and complementary selectivity to RP chromatography. Herein, we implemented
HILIC-MS for separation and quantifi cation of low molecular weight nucleotides and phospholipids in a nanoparticle-treated
lung cells and cells under at diff erent stages of hypoxia. Th e challenges, namely necessity of liquid-liquid and solid phase
extraction, samples stability, dilution re-assay and matrix eff ect in tissues are resolved and discussed. Th e elution conditions
were subsequently optimized by evaluating organic content, pH and salt concentration in the mobile phase allowing a short
simple isocratic run of only 20 min for nucleotides and 9 min for phospholipids, respectively.