LC/MS lipid profiling from human serum: a new method for global lipid extraction
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.
KeywordsLipidomics Mass spectrometry Clinical trials Diagnostic tools Dyslipidemias
Bligh & Dyer
Methanol:methyl-tert-butyl ether:chloroform method
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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