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Int. J. Mol. Sci. 2017, 18(4), 708; doi:10.3390/ijms18040708

“Bligh and Dyer” and Folch Methods for Solid–Liquid–Liquid Extraction of Lipids from Microorganisms. Comprehension of Solvatation Mechanisms and towards Substitution with Alternative Solvents

GREEN (Groupe de Recherche en Eco-Extraction de Produits Naturels), Université d’Avignon et des Pays du Vaucluse, INRA, UMR 408, GREEN, F-84000 Avignon, France
Institut de Chimie Séparative de Marcoule, F-30207 Bagnols Sur Cèze, France
Institute of Physical and Theoretical Chemistry, University of Regensburg, D-93040 Regensburg, Germany
Author to whom correspondence should be addressed.
Academic Editor: David Arráez-Román
Received: 21 February 2017 / Revised: 13 March 2017 / Accepted: 19 March 2017 / Published: 27 March 2017
(This article belongs to the Section Bioactives and Nutraceuticals)
View Full-Text   |   Download PDF [2654 KB, uploaded 28 March 2017]   |  


Bligh and Dyer (B & D) or Folch procedures for the extraction and separation of lipids from microorganisms and biological tissues using chloroform/methanol/water have been used tens of thousands of times and are “gold standards” for the analysis of extracted lipids. Based on the Conductor-like Screening MOdel for realistic Solvatation (COSMO-RS), we select ethanol and ethyl acetate as being potentially suitable for the substitution of methanol and chloroform. We confirm this by performing solid–liquid extraction of yeast (Yarrowia lipolytica IFP29) and subsequent liquid–liquid partition—the two steps of routine extraction. For this purpose, we consider similar points in the ternary phase diagrams of water/methanol/chloroform and water/ethanol/ethyl acetate, both in the monophasic mixtures and in the liquid–liquid miscibility gap. Based on high performance thin-layer chromatography (HPTLC) to obtain the distribution of lipids classes, and gas chromatography coupled with a flame ionisation detector (GC/FID) to obtain fatty acid profiles, this greener solvents pair is found to be almost as effective as the classic methanol–chloroform couple in terms of efficiency and selectivity of lipids and non-lipid material. Moreover, using these bio-sourced solvents as an alternative system is shown to be as effective as the classical system in terms of the yield of lipids extracted from microorganism tissues, independently of their apparent hydrophilicity. View Full-Text
Keywords: Bligh and Dyer; Folch; bio-sourced solvent; yeast; lipids Bligh and Dyer; Folch; bio-sourced solvent; yeast; lipids

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Breil, C.; Abert Vian, M.; Zemb, T.; Kunz, W.; Chemat, F. “Bligh and Dyer” and Folch Methods for Solid–Liquid–Liquid Extraction of Lipids from Microorganisms. Comprehension of Solvatation Mechanisms and towards Substitution with Alternative Solvents. Int. J. Mol. Sci. 2017, 18, 708.

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