Ultra-high-performance liquid chromatography electrospray ionization tandem mass spectrometry for accurate analysis of glycerophospholipids and sphingolipids in drug resistance tumor cells

Lin Li; Linlin Wang; Dihua Shangguan; Yanbo Wei; Juanjuan Han; Shaoxiang Xiong; Zhenwen Zhao

doi:10.1016/j.chroma.2015.01.013

Ultra-high-performance liquid chromatography electrospray ionization tandem mass spectrometry for accurate analysis of glycerophospholipids and sphingolipids in drug resistance tumor cells

J Chromatogr A. 2015 Feb 13:1381:140-8. doi: 10.1016/j.chroma.2015.01.013. Epub 2015 Jan 14.

Authors

Lin Li¹, Linlin Wang², Dihua Shangguan², Yanbo Wei¹, Juanjuan Han², Shaoxiang Xiong², Zhenwen Zhao³

Affiliations

¹ Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry Chinese Academy of Sciences, Beijing Mass Spectrum Center, Beijing 100190, China; Graduate School, University of Chinese Academy of Sciences, Beijing 100049, China.
² Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry Chinese Academy of Sciences, Beijing Mass Spectrum Center, Beijing 100190, China.
³ Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry Chinese Academy of Sciences, Beijing Mass Spectrum Center, Beijing 100190, China. Electronic address: zhenwenzhao@iccas.ac.cn.

PMID: 25614189
DOI: 10.1016/j.chroma.2015.01.013

Abstract

Glycerophospholipids and sphingolipids are important signaling molecules which are involved in many physiological and pathological processes. Here we reported an effective method for accurate analysis of these lipids by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The methanol method was adopted for extraction of lipids due to its simplicity and high efficiency. It was found that two subclasses of sphingolipids, sulfatide (ST) and cerebroside (CB), were heat labile, so a decreased temperature in the ion source of MS might be necessary for these compounds analysis. In addition, it was found that the isobaric interferences were commonly existent, for example, the m/z of 16:0/18:1 PC containing two (13)C isotope being identical to that of 16:0/18:0 PC determined by a unit mass resolution mass spectrometer; therefore, a baseline separation of interferential species was required to maintain selectivity and accuracy of analysis. In this work, an ultra-high-performance liquid chromatography (UHPLC)-based method was developed for separation of interferential species. Moreover, in order to deal with the characteristics of different polarity and wide dynamic range of glycerophospholipids and sphingolipids in biological systems, three detecting conditions were combined together for comprehensive and rational analysis of glycerophospholipids and sphingolipids. The method was utilized to profile glycerophospholipids and sphingolipids in drug resistant tumor cells. Our results showed that many lipids were significantly changed in drug resistant tumor cells compared to paired drug sensitive tumor cells. This is a systematic report about the isobaric interferences and heat labile compounds interferences when analyzing glycerophospholipids and sphingolipids by ESI-MS/MS, which aids in ruling out one potential source of systematic error to ensure the accuracy of analysis.

Keywords: Glycerophospholipids; MCF-7; MCF-7R; Sphingolipids; UHPLC–ESI-MS/MS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antineoplastic Agents / pharmacology
Chromatography, High Pressure Liquid / methods
Drug Resistance, Neoplasm*
Glycerophospholipids / analysis*
Humans
MCF-7 Cells
Spectrometry, Mass, Electrospray Ionization / methods
Sphingolipids / analysis*
Tandem Mass Spectrometry / methods

Substances

Antineoplastic Agents
Glycerophospholipids
Sphingolipids