Yuliya E Silina
INM-Leibniz Institute for New Materials, Germany
Title: Hydrophilic interaction chromatography-tandem mass spectrometry for the cellular analysis: opportunities and cOlimpica 2enges
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. The 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 effect 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.