“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

Abstract

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.

Effect of Sludge Concentration and Crude Glycerol Matrix as a Substrate on the Production of Single-Cell Oil by Oleaginous Yeast Yarrowia lipolytica SKY7

“…A small fraction of fermentation broth was serially diluted in 0.85% NaCl solution and used to estimate the CFU/mL by using the spread plating method. Lipid analysis was performed by extracting the lipid from the suspended solids using the chloroform-methanol extraction method [11]. The residual glycerol present in the supernatant fraction of the sample was estimated by using the spectrophotometer method as used in Kuhn, et al [12].…”

Value of hydroalcoholic treatment of rapeseed for oil extraction and protein enrichment

“…Increasing concerns for health, safety and environmental care have prompted to study alternative solvents in order to substitute hexane in oil extraction (Li et al, 2014;Sicaire et al, 2015;Baümler et al, 2016;Breil et al, 2017). Sicaire et al (2015) and Breil et al (2017) used physical properties of solvents (viscosity, boiling point, vaporization enthalpy and toxicity), theoretical solvent-lipids solubility and extraction experimentations in order to compare different solvents. They demonstrated that extraction efficiency varies depending on the lipid class or component (TAG, phospholids, tocopherols...), and the solvent choice depends on lipids composition of the meal.…”

The Correlation between MicroRNA-199a and White Adipose Tissue in C57/BL6J Mice with High-Fat Diet

“…Subcutaneous adipose tissue and visceral adipose tissue were dissected, weighed, and stored at −80°C. Lipids from subcutaneous adipose tissue were extracted by the Folch method as previously described (Breil, Abert Vian et al 2017), and the levels of FFAs and TGs were measured with the use of enzyme-linked immunoassays (Shanghai Tongwei reagent biological technology Co., Ltd., China) and the kit catalogue numbers were TWp002083 and TWp003393. The concentrations of FFAs and TGs were calculated based on a standard curve, which is made standard concentration (μmol/L) as abscissas and optical density (OD) value (lg (1/trans)) as ordinate.…”