Analytical and Bioanalytical Chemistry

, Volume 406, Issue 30, pp 7937–7948 | Cite as

LC/MS lipid profiling from human serum: a new method for global lipid extraction

  • Roberto Maria Pellegrino
  • Alessandra Di Veroli
  • Aurora Valeri
  • Laura Goracci
  • Gabriele Cruciani
Research Paper

Abstract

Over the last decade, technological advances have improved the sensitivity and selectivity of LC/MS analyzers, providing very efficient tools for lipidomics research. In particular, the nine lipid classes that constitute 99 % of the human serum lipidome (sterols, cholesteryl esters, phosphocholines, phosphoethanolamines, sphingomyelins, triacylglycerols, fatty acids, lysophosphocholines, and diacylglycerols) can be easily detected. However, until today there has not been a unique technique for sample preparation that provides a satisfactory recovery for all of these nine classes together. In this work, we have developed and validated a new one-phase extraction (OPE) method that overcomes this limitation. This method was also compared with the gold standard lipid extraction methods such as Folch, Bligh & Dyer, and recently developed methods with methanol and methyl-tert-butyl ether. Results demonstrate that the mixture of methanol/chloroform/MTBE (MMC) provides a recovery very close to 100 % for all nine lipid classes of the human serum investigated. For this extraction method, 100 μL of human serum is incubated with 2 mL of the solvents mixture, then vortexed and centrifuged. For its simplicity of execution, rapidity, reproducibility, and the reduced volume of sample required, this method opens the door to the use of human serum lipid profiling for large-scale applications in scientific research and clinical trials.

Keywords

Lipidomics Mass spectrometry Clinical trials Diagnostic tools Dyslipidemias 

Abbreviations

BD

Bligh & Dyer

FO

Folch

MA

Methanol:acetone method

MC

Methanol:chloroform method

ME

Methanol method

MIPA

Methanol:isopropanol method

MMC

Methanol:methyl-tert-butyl ether:chloroform method

OPE

One-phase extraction

SD

Standard deviation

TPE

Two-phase extraction

Notes

Acknowledgments

A.D.V. thanks Regione Umbria, this research project was partly supported by 2007–2013 ESF “Competitiveness and Employment objective” Umbrian Regional Operational  Programme (ROP), Avviso pubblico aiuti individuali per la realizzazione di progetti di ricerca, project “Correlazione tra impronta lipidica del sangue e incidenza delle malattie neurodegenerative. Sviluppo di un metodo non invasivo e poco costoso per abbattere i costi della sanità pubblica regionale”.

The authors thank the Centro Ospedaliero di Riabilitazione Intensiva (C.O.R.I.), Passignano sul Trasimeno (Perugia, Italy), for providing serum samples, and Molecular Discovery Ltd. (UK) for free software and financial support.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Roberto Maria Pellegrino
    • 1
  • Alessandra Di Veroli
    • 1
  • Aurora Valeri
    • 1
  • Laura Goracci
    • 1
  • Gabriele Cruciani
    • 1
  1. 1.Department of Chemistry, Biology, and BiotechnologiesUniversity of PerugiaPerugiaItaly

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