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.
Abstract:Lipid-based oleaginous microorganisms are potential candidates and resources for the sustainable production of biofuels. This study was designed to evaluate the performance of several alternative bio-based solvents for extracting lipids from yeasts. We used experimental design and simulation with Hansen solubility simulations and the conductor-like screening model for realistic solvation (COSMO-RS) to simulate the solubilization of lipids in each of these solvents. Lipid extracts were analyzed by high performance thin-layer chromatography (HPTLC) to obtain the distribution of lipids classes and gas chromatography coupled with a flame ionization detector (GC/FID) to obtain fatty acid profiles. Our aim was to correlate simulation with experimentation for extraction and solvation of lipids with bio-based solvents in order to make a preliminary evaluation for the replacement of hexane to extract lipids from microorganisms. Differences between theory and practice were noted for several solvents, such as CPME, MeTHF and ethyl acetate, which appeared to be good candidates to replace hexane.
In the present work, two different ways of lipids extraction from Yarrowia lipolytica yeast were investigated in order to maximize the extraction yield. Firstly, various modern techniques of extraction including ultrasound, microwave, and bead milling were tested to intensify the efficiency of lipid recovery. Secondly, several pretreatments such as freezing/defrosting, cold drying, bead milling, and microwave prior two washing of mixture solvent of chloroform:methanol (1:2, v/v) were study to evaluate the impact on lipid recovery. All these treatments were compared to conventional maceration, in terms of lipids extraction yield and lipid composition analysis. The main result of this study is the large difference of lipid recovery among treatments and the alteration of lipids profile after microwave and ultrasound techniques.
There is a great interest in finding alternatives and green solvents in extraction processes to replace petroleum based solvents. In order to investigate these possibilities, computational methods, as Hansen solubility parameters (HSP) and conductor-like screening model for real solvent (COSMO-RS), were used in this work to predict the solvation power of a series of solvents in salmon fish lipids. Additionally, experimental studies were used to evaluate the performance in lipids extraction using 2-methyltetrahydrofurane, cyclopentyl methyl ether, dimethyl carbonate, isopropanol, ethanol, ethyl acetate, p-cymene and d-limonene compared with hexane. Lipid classes of extracts were obtained by using high performance thin-layer chromatography (HPTLC), whereas gas chromatography with a flame ionization detector (GC/FID) technique was employed to obtain fatty acid profiles. Some differences between theoretical and experimental results were observed, especially regarding the behavior of p-cymene and d-limonene, which separate from the predicted capability. Results obtained from HPTLC indicated that p-cymene and d-limonene extract triglycerides (TAGs) and diglycerides (DAGs) at levels of 73 and 19%, respectively, whereas the other studied extracts contain between 75 and 76% of TAGs and between 16 and 17% of DAGs. Fatty acid profiles, obtained by using GC-FID, indicated that saturated fatty acids (SFAs) between 19.5 and 19.9% of extracted oil, monounsaturated fatty acids (MUFAs) in the range between 43.5 and 44.9%, and PUFAs between 31.2 and 34.6% were extracted. p-Cymene and limonene extracts contained lower percentages than the other studied solvents of some PUFAs due probably to the fact that these unsaturated fatty acids are more susceptible to oxidative degradation than MUFAs. Ethyl acetate has been found to be the best alternative solvent to hexane for the extraction of salmon oil lipids. Graphical Abstract ᅟ.
Biorefining
of bilberry pomace using innovative technologies such
as microwave hydrodiffusion and gravity extraction (MHG) and ultrasound-assisted
extraction (UAE) with different concentrations of ethanol/water as
the solvent was established. Bead milling was also utilized in this
sequential extraction scheme to remove the lipophilic fraction. The
solubility index of target polyphenols was predicted using a computational
tool (COSMO-RS) and compared to experimental results obtained by in
vitro antioxidant activity assessments. MHG extracts obtained using
microwave power of 2 W/g had the highest Folin–Ciocalteu reducing
capacity (43.46 ± 0.48 mg gallic acid equivalent (GAE)/g of extract),
total flavonoid (4.17 ± 0.04 mg quercetin equivalents (QE)/g
of extract), total monomeric anthocyanin content (12.19 ± 0.13
mg delphinidin-3-O-glucoside equivalents (D3GE)/g
of extract), and radical scavenging capacity (22.64 ± 2.23 mg
trolox equivalents (TE)/g of extract). In UA ethanol/water extracts,
the highest flavonoid (10.41 ± 0.08 mg QE/g of extract) and anthocyanin
content (12.19 ± 0.51 D3GE mg/g of extract) was present in ethanol
(100%), these results were in good correlation with computational
predictions. The lipid fraction recovered from pomace using bead-milling
extraction was mainly composed of oil rich in polyunsaturated linoleic
and linolenic acids.
This study was performed to compare, in term of quality and quantity, conventional and green lipid extraction procedures of three Tunisian date palm (Phoenix dactylifera L.) seeds cultivars (Deglet Nour, Allig and Belah). Extractions were performed using n-hexane and 2-methyltetrahydrofuran (MeTHF) as solvents. Identification and quantification of extracted fats and lipids were carried out by gas chromatography coupled with a flame ionization detector and high performance thin-layer chromatography. Results showed that Deglet Nour presents the highest lipids extraction yield (7.24-5.97%) using the both solvents. Hansen solubility simulation demonstrated that MeTHF is a good alternative solvent for lipid date seeds extraction. In addition, extraction of oils from date pits with ultrasound and microwave were better in terms of extraction time (30 min versus 8 h) comparing to soxhlet procedure and of extraction yield comparing to maceration (about 6% versus 4%). Triglycerides represent almost the whole recovered oils composition with 99% and the fatty acids were mainly oleic (44.02-46.9%), lauric (20.00-23.10%), myristic (8.88-11.26%), palmitic (9.00-10.73%) and linoleic (6.13-9.21%) acids.
Near-infrared (NIR) spectroscopy was evaluated as a rapid method for the determination of oleic, palmitic, linoleic and linolenic acids as well as omega-3, omega-6, and to predict polyunsaturated, monounsaturated and saturated fatty acids, together with triacylglycerides, diglycerides, free fatty acids and ergosterol in salmon oil. To do it, Partial Least Squares (PLS) regression models were applied to correlate NIR spectra with aforementioned fatty acids and lipid classes. Results obtained were validated in front of reference procedures based on high performance thin layer and gas chromatography. PLS-NIR has a good predictive capability with relative root mean square error of prediction (RRMSEP) values below or equal to 1.8% and provides rapid analysis without the use of any chemicals making it an environmentally friendly methodology.
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