Indigenous microalgae strains characterization for a sustainable biodiesel production.

Microalgae have been widely recognized as a promising feedstock for sustainable biofuels production to tackle global warming and pollution issues related to fossil fuels uses. This study identified and analyzed indigenous microalgae strains for biodiesel production, specifically Chlorella vulgaris and Coelastrella thermophila var. globulina, from two distinct locations in Algeria. Molecular identification confirmed their identity, and the microalgae exhibited notable growth characteristics. Local Chlorella vulgaris and Coelastrella thermophila var. globulina showed good growth and high biomass yield, compared to Chlorella vulgaris CCAP211/11B reaching a weight of 1.48 g L-1 , 1.95 g L-1 , and 2.10 g L-1 , respectively. Lipids content of local Chlorella vulgaris, Coelastrella thermophila var. globulina, and Chlorella vulgaris CCAP211/11B, were found to be 31.39 ± 3.3%, 17 ± 2.26%, and 19 ± 0.64%, respectively. Chlorella vulgaris stood out as a candidate for biodiesel production due to its equilibrium between SFA and PUFA (43.24% and 45.27%). FAs are predominated by SFA and MUFA for Coelastrella thermophila var. globulina with value of 81.49% (SFA+MUFA). Predicted biodiesel qualities comply with ASTM6751 and EN14214 standards. Studied microalgae have therefore a promising potential for biodiesel production. However, optimising cultivation conditions is necessary to enhance biomass and lipids yield at a large scale.

Towards Substitution of Hexane as Extraction Solvent of Food Products and Ingredients with No Regrets.

Hexane is a solvent used extensively in the food industry for the extraction of various products such as vegetable oils, fats, flavours, fragrances, colour additives or other bioactive ingredients. As it is classified as a "processing aid", it does not have to be declared on the label under current legislation. Therefore, although traces of hexane may be found in final products, especially in processed products, its presence is not known to consumers. However, hexane, and in particular the n-hexane isomer, has been shown to be neurotoxic to humans and has even been listed as a cause of occupational diseases in several European countries since the 1970s. In order to support the European strategy for a toxic-free environment (and toxic-free food), it seemed important to collect scientific information on this substance by reviewing the available literature. This review contains valuable information on the nature and origin of the solvent hexane, its applications in the food industry, its toxicological evaluation and possible alternatives for the extraction of natural products. Numerous publications have investigated the toxicity of hexane, and several studies have demonstrated the presence of its toxic metabolite 2,5-hexanedione (2,5-HD) in the urine of the general, non-occupationally exposed population. Surprisingly, a tolerable daily intake (TDI) has apparently never been established by any food safety authority. Since hexane residues are undoubtedly found in various foods, it seems more than necessary to clearly assess the risks associated with this hidden exposure. A clear indication on food packaging and better information on the toxicity of hexane could encourage the industry to switch towards one of the numerous other alternative extraction methods already developed.

2-Methyloxolane (2-MeOx) as Sustainable Lipophilic Solvent to Substitute Hexane for Green Extraction of Natural Products. Properties, Applications, and Perspectives.

This review presents a complete picture of current knowledge on 2-methyloxolane (2-MeOx), a bio-based solvent for the extraction of natural products and food ingredients. It provides the necessary background about the properties of 2-MeOx, not only its solvent power and extraction efficiency, but its detailed toxicological profile and environmental impacts are discussed. We compared 2-MeOx with hexane which is the most used petroleum-based solvent for extraction of lipophilic natural products. The final part focuses on successful industrial transfer, including technologic, economic, and safety impacts. The replacement of petroleum-based solvents is a hot research topic, which affects several fields of modern plant-based chemistry. All the reported applications have shown that 2-MeOx is an environmentally and economically viable alternative to conventional petroleum-based solvents for extraction of lipophilic foodstuff and natural products.

Ultrasound and deep eutectic solvents: An efficient combination to tune the mechanism of steviol glycosides extraction.

Ultrasound-assisted extraction is widely recognized as an eco-friendly technique due to low solvent consumption and time extraction as well as enhanced extraction efficiency with respect to conventional methods. Nevertheless, it would be convenient to avoid the usually used organic solvents to reduce the environment pollution. In this regard, Deep Eutectic Solvents (DES) represent nowadays a green and sustainable alternative for the extraction of bioactive compounds from natural sources. In this study, an efficient extraction of stevioside and rebaudioside A from Stevia rebaudiana coupling ultrasound with DES was developed. A solvent screening was performed using the predictive approach COnductor-like Screening MOdel for Real Solvent (COSMO-RS). The effect of three independent variables, namely % of water, temperature, and sonication amplitude, were investigated by the response surface methodology (RSM). Comparing ultrasound-assisted extraction (UAE) with conventional extraction, it has been demonstrated that the amount of steviol glycosides through UAE is almost three times higher than that obtained by the conventional method. Possible physicochemical factors involved in the UAE mechanism were discussed.

Review of Alternative Solvents for Green Extraction of Food and Natural Products: Panorama, Principles, Applications and Prospects.

In recent years, almost all extraction processes in the perfume, cosmetic, pharmaceutical, food ingredients, nutraceuticals, biofuel and fine chemical industries rely massively on solvents, the majority of which have petroleum origins. The intricate processing steps involved in the industrial extraction cycle makes it increasingly difficult to predict the overall environmental impact; despite the tremendous energy consumption and the substantial usage of solvents, often the yields are indicated in decimals. The ideal alternative solvents suitable for green extraction should have high solvency, high flash points with low toxicity and low environmental impacts, be easily biodegradable, obtained from renewable (non-petrochemical) resources at a reasonable price and should be easy to recycle without any deleterious effect to the environment. Finding the perfect solvent that meets all the aforementioned requirements is a challenging task, thus the decision for the optimum solvent will always be a compromise depending on the process, the plant and the target molecules. The objective of this comprehensive review is to furnish a vivid picture of current knowledge on alternative, green solvents used in laboratories and industries alike for the extraction of natural products focusing on original methods, innovation, protocols, and development of safe products.

"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.

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.

Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review.

This review presents a complete picture of current knowledge on ultrasound-assisted extraction (UAE) in food ingredients and products, nutraceutics, cosmetic, pharmaceutical and bioenergy applications. It provides the necessary theoretical background and some details about extraction by ultrasound, the techniques and their combinations, the mechanisms (fragmentation, erosion, capillarity, detexturation, and sonoporation), applications from laboratory to industry, security, and environmental impacts. In addition, the ultrasound extraction procedures and the important parameters influencing its performance are also included, together with the advantages and the drawbacks of each UAE techniques. Ultrasound-assisted extraction is a research topic, which affects several fields of modern plant-based chemistry. All the reported applications have shown that ultrasound-assisted extraction is a green and economically viable alternative to conventional techniques for food and natural products. The main benefits are decrease of extraction and processing time, the amount of energy and solvents used, unit operations, and CO2 emissions.

Microwave, ultrasound, thermal treatments, and bead milling as intensification techniques for extraction of lipids from oleaginous Yarrowia lipolytica yeast for a biojetfuel application.

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.

Ultrasound induced green solvent extraction of oil from oleaginous seeds.

Ultrasound-assisted extraction of rapeseed oil was investigated and compared with conventional extraction for energy efficiency, throughput time, extraction yield, cleanness, processing cost and product quality. A multivariate study enabled us to define optimal parameters (7.7 W/cm(2) for ultrasonic power intensity, 40 °C for processing temperature, and a solid/liquid ratio of 1/15) for ultrasound-assisted extraction of oil from oilseeds to maximize lipid yield while reducing solvent consumption and extraction time using response surface methodology (RSM) with a three-variable central composite design (CCD). A significant difference in oil quality was noted under the conditions of the initial ultrasound extraction, which was later avoided using ultrasound in the absence of oxygen. Three concepts of multistage cross-current extraction were investigated and compared: conventional multistage maceration, ultrasound-assisted maceration and a combination, to assess the positive impact of using ultrasound on the seed oil extraction process. The study concludes that ultrasound-assisted extraction of oil is likely to reduce both economic and ecological impacts of the process in the fat and oil industry.

Bio-Based Solvents for Green Extraction of Lipids from Oleaginous Yeast Biomass for Sustainable Aviation Biofuel.

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.

Effects of Temperature and Other Operational Parameters on Chlorella vulgaris Mass Cultivation in a Simple and Low-Cost Column Photobioreactor.

Mass production of microalgae worldwide, and even more so in developing countries, is strongly contingent upon the availability of economical and efficient photobioreactors (PBRs) that are amenable for use in resource-limited environments. Such options are limited. This work assesses the effects of temperature, CO2 enrichment, and mixing by air + CO2 bubbling on Chlorella vulgaris biomass production in a simple, low-cost 84-L column PBR. Cultivation at 25, 30, and 35 °C in a batch process showed that biomass production was negatively affected above 30 °C. Specific growth rates at each temperature were 0.75, 0.76, and 0.63 day(-1), respectively, with batch productivities of 70.50, 81.67, and 35.83 mg L(-1) day(-1). While a relatively low CO2/air ratio (1 %) seemed beneficial during the early stages of cultivation, higher concentrations were required to maintain growth rate and achieve higher biomass concentrations around 1000 mg L(-1). Cultivation with air + CO2 bubbling rates of 100, 200, and 400 L h(-1) led to specific growth rates (and batch productivities) of 0.64 day(-1) (59.58 mg L(-1) day(-1)), 0.74 day(-1) (81.67 mg L(-1) day(-1)), and 0.80 day(-1) (86.67 mg L(-1) day(-1)), respectively. The results indicate that high biomass productivities of C. vulgaris can be obtained up to 30 °C with moderate (2 %) to high (10 %) CO2 in a fairly simple PBR.

Alternative bio-based solvents for extraction of fat and oils: solubility prediction, global yield, extraction kinetics, chemical composition and cost of manufacturing.

The present study was designed to evaluate the performance of alternative bio-based solvents, more especially 2-methyltetrahydrofuran, obtained from crop's byproducts for the substitution of petroleum solvents such as hexane in the extraction of fat and oils for food (edible oil) and non-food (bio fuel) applications. First a solvent selection as well as an evaluation of the performance was made with Hansen Solubility Parameters and the COnductor-like Screening MOdel for Realistic Solvation (COSMO-RS) simulations. Experiments were performed on rapeseed oil extraction at laboratory and pilot plant scale for the determination of lipid yields, extraction kinetics, diffusion modeling, and complete lipid composition in term of fatty acids and micronutrients (sterols, tocopherols and tocotrienols). Finally, economic and energetic evaluations of the process were conducted to estimate the cost of manufacturing using 2-methyltetrahydrofuran (MeTHF) as alternative solvent compared to hexane as petroleum solvent.

New procedure for extraction of algal lipids from wet biomass: a green clean and scalable process.

A new procedure, called Simultaneous Distillation and Extraction Process (SDEP), for lipid extraction from wet microalgae (Nannochloropsis oculata and Dunaliella salina) was reported. This method does not require a pre-drying of the biomass and employs alternative solvents such as d-limonene, α-pinene and p-cymene. This procedure has been compared with Soxhlet extraction (Sox) and Bligh & Dyer method (B&D). For N. oculata, results showed that SDEP-cymene provided similar lipid yields to B&D (21.45% and 23.78%), while SDEP-limonene and pinene provided lower yields (18.73% and 18.75% respectively). For D. salina, SDEP-pinene provided the maximum lipid yield (3.29%) compared to the other solvents, which is quite close to B&D result (4.03%). No significant differences in terms of distribution of lipid classes and fatty acid composition have been obtained for different techniques. Evaluation of energy consumption indicates a substantial saving in the extraction cost by SDEP compared to the conventional extraction technique, Soxhlet.

Green extraction of natural products: concept and principles.

The design of green and sustainable extraction methods of natural products is currently a hot research topic in the multidisciplinary area of applied chemistry, biology and technology. Herein we aimed to introduce the six principles of green-extraction, describing a multifaceted strategy to apply this concept at research and industrial level. The mainstay of this working protocol are new and innovative technologies, process intensification, agro-solvents and energy saving. The concept, principles and examples of green extraction here discussed, offer an updated glimpse of the huge technological effort that is being made and the diverse applications that are being developed.

Terpenes as green solvents for extraction of oil from microalgae.

Herein is described a green and original alternative procedure for the extraction of oil from microalgae. Extractions were carried out using terpenes obtained from renewable feedstocks as alternative solvents instead of hazardous petroleum solvents such as n-hexane. The described method is achieved in two steps using Soxhlet extraction followed by the elimination of the solvent from the medium using Clevenger distillation in the second step. Oils extracted from microalgae were compared in terms of qualitative and quantitative determination. No significant difference was obtained between each extract, allowing us to conclude that the proposed method is green, clean and efficient.

"Solvent-free" ultrasound-assisted extraction of lipids from fresh microalgae cells: a green, clean and scalable process.

In order to comply with criteria of green chemistry concepts and sustainability, a new procedure has been performed for solvent-free ultrasound-assisted extraction (UAE) to extract lipids from fresh Nannochloropsis oculata biomass. Through response surface methodology (RSM) parameters affecting the oil recovery were optimized. Optimum conditions for oil extraction were estimated as follows: (i) 1000 W ultrasonic power, (ii) 30 min extraction time and (iii) biomass dry weight content at 5%. Yields were calculated by the total fatty acids methyl esters amounts analyzed by GC-FID-MS. The maximum oil recovery was around 0.21%. This value was compared with the one obtained with the conventional extraction method (Bligh and Dyer). Furthermore, effect of temperature on the yield was also investigated. The overall results show an innovative and effective extraction method adapted for microalgae oil recovery, without using solvent and with an enable scaling up.

Combined extraction processes of lipid from Chlorella vulgaris microalgae: microwave prior to supercritical carbon dioxide extraction.

Extraction yields and fatty acid profiles from freeze-dried Chlorella vulgaris by microwave pretreatment followed by supercritical carbon dioxide (MW-SCCO(2)) extraction were compared with those obtained by supercritical carbon dioxide extraction alone (SCCO(2)). Work performed with pressure range of 20-28 Mpa and temperature interval of 40-70 °C, gave the highest extraction yield (w/w dry weight) at 28 MPa/40 °C. MW-SCCO(2) allowed to obtain the highest extraction yield (4.73%) compared to SCCO(2) extraction alone (1.81%). Qualitative and quantitative analyses of microalgae oil showed that palmitic, oleic, linoleic and α-linolenic acid were the most abundant identified fatty acids. Oils obtained by MW-SCCO(2) extraction had the highest concentrations of fatty acids compared to SCCO(2) extraction without pretreatment. Native form, and microwave pretreated and untreated microalgae were observed by scanning electronic microscopy (SEM). SEM micrographs of pretreated microalgae present tearing wall agglomerates. After SCCO(2), microwave pretreated microalgae presented several micro cracks; while native form microalgae wall was slightly damaged.

Clean recovery of antioxidant flavonoids from onions: optimising solvent free microwave extraction method.

A solvent free microwave hydrodiffusion and gravity extraction (MHG) of flavonol content from onion (Allium cepa L.) was studied. Effectiveness of this innovative method in extraction of onion total phenolic content, total quercetin (TQ), quercetin aglycon (QA), quercetin-3,4'-diglucoside (QDG), quercetin-4'-monoglucoside (Q4G), quercetin-3-monoglucoside (Q3G), kaempferol (KMF) and myricetin (MRT) have been evaluated and compared with conventional solvent extraction. Microwave extraction offers important advantages like shorter extraction time (23min), cleaner feature (no solvent or water used) and extraction of valuable onion crude juice retaining fresh organoleptic properties with higher phenolic content (58.29mgGAE/gDW) at optimized power (500W). Microwave extraction resulted significant yield (81.5%) with 41.9% of flavonol contents, with better retain of remaining flavonoids (55.9%) in residues of onions. QDG (239.7mg/100gDW) and Q4G (82.55mg/100gDW) have been reported the main flavonol in this study. Minor quantities of QA (traces), Q3G (4.22mg/100gDW) and KMF (3.99mg/100gDW) were also detected in microwave onion extracts.

Improved microwave steam distillation apparatus for isolation of essential oils. Comparison with conventional steam distillation.

Steam distillation (SD) is routinely used by analysts for the isolation of essential oils from herbs, flowers and spices prior to gas chromatographic analysis. In this work, a new process design and operation for an improved microwave steam distillation (MSD) of essential oils from aromatic natural products was developed. To demonstrate its feasibility, MSD was compared with the conventional technique, SD, for the analysis of volatile compounds from dry lavender flowers (Lavandula angustifolia Mill., Lamiaceae). Essential oils isolated by MSD were quantitatively (yield) and qualitatively (aromatic profile) similar to those obtained by SD, but MSD was better than SD in terms of rapidity (6 min versus 30 min for lavender flowers), thereby allowing substantial savings of costs in terms of time and energy. Lavender flowers treated by MSD and SD were observed by scanning electron microscopy. Micrographs provide evidence of more rapid opening of essential oil glands treated by MSD, in contrast to conventional SD.

Microwave hydrodiffusion and gravity, a new technique for extraction of essential oils.

A new process design and operation for the extraction of essential oils was developed. Microwave hydrodiffusion and gravity (MHG) is a combination of microwaves for hydrodiffusion of essential oils from the inside to the exterior of biological material and earth gravity to collect and separate. MHG is performed at atmospheric pressure without adding any solvent or water. MHG has been compared with a conventional technique, hydrodistillation (HD), for the extraction of essential oil from two aromatic herbs: spearmint (Mentha spicata L.) and pennyroyal (Mentha pulegium L.) belonging to the Labiatae family. The essential oils extracted by MHG for 15 min were quantitatively (yield) and qualitatively (aromatic profile) similar to those obtained by conventional hydrodistillation for 90 min. MHG also prevents pollution through potential 90% of energy saved which can lead to greenhouse gas emission benefits.