Fuel Processing Technology 193 (2019) 295–303

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Fuel Processing Technology

journal homepage: www.elsevier.com/locate/fuproc

Review paper

Extraction techniques in sustainable biofuel production: A concise review T

a,⁎ b b
Peng Li , Kiyoshi Sakuragi , Hisao Makino
a
Environmental Science Research Laboratory, Central Research Institute of Electric Power Industry (CRIEPI), 1646 Abiko, Chiba-ken, 270-1194, Japan
b
Energy Engineering Research Laboratory, Central Research Institute of Electric Power Industry (CRIEPI), Yokosuka, Kanagawa-Ken 240-0196, Japan

ARTICLE INFO ABSTRACT

Keywords: The concerns of fossil fuel depletion and environmental issues have increased general interest in studies on the
Extraction development of sustainable biofuel. In the biofuel production process, the extraction technique is a key tech-
Sustainable energy nology with respect to energy consumption and product value. In this review, recent advances related to the
Biofuel extraction process in biofuel production are discussed. The major extraction methods were classified as con-
Biomass
ventional solvent extraction (CSE), physical-supported solvent extraction (PSSE), supercritical fluid extraction
Algae biofuel
(SFE), and novel extractions. We address each of these classified extraction methods in turn both in relation to
the feedstock utilized in the different biofuel generation methods and to the targeted biofuels. Our objective is to
provide a concise, timely, and comprehensive review on various extraction methods applied/studied regarding
biofuel production, their principles and advantages/limitations, and recent case studies. In particular, the
classified extraction methods in the production of third-generation algal biofuel were intensively evaluated.
Consequently, the concept of free from drying and cell disruption (FDC) routes was advanced herein as it may
possess the potential to reduce the requirement for input energy during the production of algae biofuels.

1. Introduction esterification), and thermochemical (e.g. gasification, liquefaction)

approaches. In particular, thermochemical processes can be used to
Fossil fuels still serve as the primary global energy resource and produce oil and gas [5,6] whereas biochemical processes yield products
account for more than 88% of the primary energy consumption [1], such as bioethanol. These conversion processes have been described in
although new sources of cheap fossil fuels are no longer available and detail in several recent papers (e.g. [7–9]). Within the physico-chemical
experts have issued warnings regarding the possible depletion of the approach, extraction allows for recovery of the desired compositions or
current sources in the near future [2]. In addition, greenhouse gas removal of undesired substances from target feedstock [10].
emissions such as CO2 caused by the combustion of fossil fuels also The goal of the current review was to summarize recent studies of
represent a serious concern. Therefore, the development of sustainable extraction methods and classify these methods as well as to discuss their
energy resources and reduction of the greenhouse gas substances from possible application in biofuel production in relation to the feedstocks
fossil fuels have become essential topics of interest worldwide [3]. The utilized for the different biofuel generation strategies. Notably, classi-
main current sustainable energy resources consist of the energy that can fication of the extraction methods used in biofuel production is com-
be derived from biomass and natural phenomena such as solar and wind plex, especially considering the large volume of information available
(Fig. 1). The latter comprise the two largest sources of renewable regarding this topic. This review was thus structured around the main
electricity and have the major advantage that the source of energy is theme “classification of existing extraction methods” and is organized
free albeit the major disadvantage of being variable and intermittent. as follows. Section 2 illustrates the classification and basic information
In turn, biomasses constitute the third primary energy source regarding extraction methods. Section 3 presents the generation of
overall after coal and oil [4]. Many technological options to convert biofuel and the extraction methods applied or studied with regard to
biomass to biofuel have been and are being studied and implemented in the feedstocks of these different biofuels. Particularly, the extraction
practice with different degrees of success. Fig. 1 describes the general studies related to the new generation of algae biofuels are intensively
options for biofuel derivation from biomasses. Biomasses can be con- reviewed. The meaningful considerations for selection of extraction
verted into fuel products by means of biological (e.g. fermentation, methods based on algae biofuel production are presented in Section 4 as
anaerobic digestion), physico-chemical (e.g. extraction, trans- this is believed to likely comprise the biofuel of the future and to exhibit

⁎
Corresponding author.
E-mail address: [email protected] (P. Li).

https://doi.org/10.1016/j.fuproc.2019.05.009
Received 1 February 2019; Received in revised form 1 May 2019; Accepted 8 May 2019
Available online 28 May 2019
0378-3820/ © 2019 Elsevier B.V. All rights reserved.
P. Li, et al. Fuel Processing Technology 193 (2019) 295–303

Fig. 1. Schematic diagram of sustainable energy options.

considerable potential to deliver clean energy. Finally, Section 5 pro- substances such as lipids [18] and hydrocarbon from various biomasses
vides the main conclusions and highlights of the current review. [19]. Multifarious solvents can be selected with solvent selection
usually based on the type of biomass and the specific components
contained therein. Chemical and physical properties such as polarity
2. Basic information and classification of extraction methods comprise the main factors influencing the efficiency and selectivity of
extraction solvents in addition to the extraction conditions [20]. In
2.1. Basic information on extraction techniques addition, economic and environmental concerns are also important is-
sues that must be considered in the application of solvent extraction
Extraction normally constitutes a midstream step in the physico- [21,22]. Some representative extractable feedstocks for the production
chemical process of biofuel production (Fig. 2) as some procedures such of different generations of biofuels are listed in Table 1. The biofuel
as cultivation and drying should be performed prior to extraction (up- products acquired via extraction processes mainly comprise the pre-
stream), whereas the extracts themselves need to be further refined by cursors of bio-diesel and jet fuel. In some cases, bio-alcohols can also be
using procedures such as fractional distillation and trans-esterification acquired from the extracts of wood biomasses. Moreover, bio-solid fuel
(downstream). can be acquired after removal of the water and impurities from bio-
Extraction can be basically divided into mechanical, physical, and masses by using extraction (dewatering) technologies.
chemical approaches. The representative methods in the current clas- Notably, a general search will yield hundreds of thousands of links
sification are listed in Fig. 2; the respective theoretical backgrounds involving information regarding either or both extraction methods or
available in the cited references [11–17] and described as relevant biofuels. However, even within academic-technical review papers that
hereafter. All three approaches are used in biofuel production although concentrate on biofuel studies, the content directly related to extraction
the latter two are considered the more important and are frequently only represents a very limited part of the article; moreover, recent
applied. Solvents in different forms are widely used to extract fueling

Fig. 2. Schematic diagram of classification of extraction methods. SFE, supercritical fluid extraction.

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Table 1
List of extractable components for the production of different generations of biofuels.
Generation Extraction methods Representative feedstocks Extractable components Targeted biofuels Ref.

First CSE Energy crops, and animal fats Fatty acids Diesel [23]
Second Mechanical, CSE, PSSE, SFE, Novel Industrial, civil, and agricultural residues Fatty acids Diesel [22,24]
Hydrocarbons Jet fuel [25]
Bio-solids Solid fuel [26]
Woody crops: oil and seeds Fatty acids Diesel [27]
Third CSE, PSSE, SFE, Novel Microalgae Lipids Diesel [28]
Hydrocarbons Jet fuel [29]

CSE, conventional solvent extraction; PSSE, physical-supported solvent extraction; SFE, supercritical fluid extraction.

Table 2
Case analysis of classified extraction methods in algae biofuel production.
E
Extraction method Representative methods Necessity of drying and cell disruption Energy demand: Ref.
Comments: C

E
CSE Polar solvent Both :Energy intensive [30]
C
(e.g. Bligh-Dyer, Methanol-chloroform) : Environmental issues etc.
E
Non-polar solvent Both :Energy intensive [30]
C
(e.g. Soxhlet with hexane) : Environmental issues etc.
E
PSSE Microwave supported Drying : Energy intensive [31]
C
: Easy to scale up
L
:Redundant energy
E
Sonication supported Drying : Energy intensive [31]
C
: Cells disrupted
simultaneously
E
SFE SCCO2 Both combined by lyophilization : High pressure high energy demand [32]
C
: Good selectivity but safety issues
E
SuCH2O Both combined by lyophilization :Energy intensive [35]
C
: Multiple products
E
Novel Switchable solvent extraction Both combined by lyophilization :Low energy consuming [38]
C
:Low extraction rate
E
Liquefied gas Neither : Low energy consuming [18]
C
:Under development

studies related to extraction have generally focused only on new tech- 2.3. Conventional solvent extraction (CSE)
nical developments. Therefore, a critical literature review devoted to
the comprehensive application, recent research, and new methods of CSE was the most frequently mentioned extraction method among
extraction in the biofuel production processes is appropriate. the selected articles. This likely occurred because as a historical
method, chemical solvent extraction has been applied in many fields
such as the food and pharmaceutical industry, and many classical CSE
2.2. Classification of extraction methods methods have been verified. Chemical solvents used for CSE are nor-
mally in the liquid form at ambient pressure and room temperature and
The general classification of extraction methods that are used in possess high selectivity and solubility for the desired compositions.
biofuel production is depicted in Fig. 2. Mechanical methods are con- Herein, some conventional and classical solvent extraction methods
ventional for first generation biofuel production, avoiding the use of such as Soxhlet extraction and Bligh-Dyer methods, which are still
chemicals while providing advantages such as the production of in- frequently applied for the separation of oil from solid materials [16,17],
stantly consumable crude oil and low equipment cost [11]. In parti- are assigned as representing CSE. A wide range of solvents can be se-
cular, the use of presses/expellers constitutes the most traditional me- lected for the Soxhlet technique individually or as mixtures including
chanical oil extraction method applied for oleaginous material. hexane, chloroform, and methanol [16]. In the Bligh-Dyer method, the
Conversely, such mechanical extraction methods are rarely utilized for combination of utilized solvents can be changed based on the polarity
the production of second- and third-generation biofuels. Physical ex- of lipids present in the target materials [17], in accordance with the
traction methods include sonication, microwaving, homogenizing, and traditional classification of organic solvents as non-polar, aprotic polar,
heating. Chemical extraction basically utilizes various solvents to ex- and protic polar based on their ability to form hydrogen bonds. Con-
tract the biofuel component from target feedstocks, comprising the sistent with this, the use of various organic solvents has been reported
well-utilized Soxhlet extractor with different solvents and the classical for biofuel production. The principle of CSE in the extraction of biofuel
Bligh-Dyer method as well as traditional liquid-liquid extraction. Unlike is that upon contact of the organic solvent with the biomass, the target
mechanical methods, in many cases the application of physical and compositions are extracted into the organic solvent owing to the per-
chemical extraction methods is combined, with the physical methods meability effect of the solvent on the biomass. Therefore, a suitable
usually being applied to support chemical solvent extraction. Therefore, organic solvent should possess biocompatibility and maximum solubi-
to provide a clear yet comprehensive assessment of these processes, lity for the target compositions. However, a notable shortcoming of CSE
physical and chemical methods were sub-classified in this review into is that most organic solvents cause health hazards and environmental
conventional solvent extraction (CSE), physical-supported solvent ex- pollution. Moreover, in CSE a pre-treatment is normally required for
traction (PSSE), and supercritical fluid extraction (SFE). In addition, extracting the available components, thereby increasing the overall cost
some new extraction methods are assigned as belonging to “novel” burden (Table 2).
methods.

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Fig. 3. Relationships between feedstocks of different generation biofuels with the classified extraction methods along with targeted biofuels.

2.4. Physical-supported solvent extraction (PSSE) to the high operational costs (Table 2) and safety-related issues; for
example, the operating temperature and pressure of supercritical CO2
In this review, CSEs supported by physical methods, such as the are above 304.25 K and 7.39 MPa, respectively. To reduce associated
most frequently used microwave and sonication methods, were as- costs, recent studies have therefore attempted to instead use near- or
signed as PSSE. Microwaves, referring to a frequency of electromagnetic sub-critical solvents such as water [35].
radiation around 0.3–300 GHz, are generally applied at a small scale for
the disruption of plant cells [13]. In turn, the application of sonication
2.6. Novel methods
for plant cell disruption in association with oil extraction has been
studied for many years. Ultrasonic cavitation is significantly more in-
Some new extraction methods that do not fit within the previously
tense at low frequency (around 18–50 kHz), with the type of plant cell
described categories were assigned as “novel” methods. For example,
wall and medium conditions such as viscosity and temperature affecting
multiple ionic liquids were assessed for their ability to extract branched
sonication efficiency [14].
hydrocarbons from an aqueous medium [29,36]. Another study de-
Notably, PSSE represents a means to integrate pre-treatment pro-
scribed a lipid extraction method using ozonation, which can produce
cesses into CSE. In general, the use of PSSE is dependent on the type of
saturated hydrocarbons from Dunaliella salina (green alga) [37].
biomass, as some require pre-treatment prior to biofuel extraction. For
Moreover, solvents with switchable hydrophilicity have been used for
example, microwaving generates high frequency waves, which may
biofuel extraction from microalgae [38]. Finally, the simultaneous ex-
destroy the biomass through shock induction. Therefore, it was recently
traction and dewatering of biofuel from wet biomasses using liquefied
suggested to represent an efficient method for disrupting oil-containing
gases such as dimethyl ether (DME) as the extraction solvent have also
plant cells [30]. Furthermore, sonication has been extensively used for
been reported [26].
microbial cells, as it disrupts both the cell wall and membrane through
the cavitation effect. The characteristics of microwaving and sonication
are shown in Table 2. 3. Extraction methods applied to different biofuel feedstocks

3.1. Extraction methods applied to the feedstocks in first-generation biofuels

2.5. Supercritical fluid extraction (SFE)
The figures of extractable feedstocks in different generations of
SFE was firstly developed for the extraction of oil in 1980 [15]. biofuels are depicted in Table 1. First-generation biofuels are produced
Although SFE also belongs to the categories of both physical and che- directly from arable food crops or animal fats. The biofuel is ultimately
mical extraction, owing to its importance, SFE is treated as independent derived from the starch, sugar, animal fat, and vegetable oil that these
from PSSE in the current review. SFE has a unique advantage compared crops or animals provide. Corn, wheat, sugar cane, and oily plants are
to CSE as it shows high selectivity (Table 2). SFE involves the use of the most commonly used first-generation biofuel feedstocks [39] with
fluid in a supercritical state as an extracting solvent, the physical and bioethanol and biodiesel being the two major types of derived first-
thermal properties of which are between those of pure liquid and gas generation biofuels. Biodiesel (fatty acid methyl ester) in particular is
[33]. Carbon dioxide (CO2) and water are the most commonly used produced through the transesterification of oils extracted from oily
supercritical fluids, which could be potentially used for biofuel pro- crops.
duction. For example, supercritical CO2 has several advantages, espe- As shown in Fig. 3, the application of extraction methods for the
cially for the extraction of low polar chemicals such as lipids from feedstocks of first-generation biofuels mainly relies upon mechanical
biomasses [34]. The disadvantages of current SFE methods are related methods, because biodiesel derived from vegetable oil accounts for a

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large proportion of first-generation biofuel and the extraction of vege- in 24.0, 18.0, and 16.0 wt% total bio-oil extraction [27]. Hexane,
table oil from seeds or plant parts is traditionally accomplished by acetone, and petroleum ether were also evaluated for the extraction of
mechanical pressing or expression [40,41]. First-generation biofuels Bauhinia monandra (orchid tree) seed oil, with petroleum ether yielding
have therefore been considered to exhibit economic and environmental 14.8 wt% oil [44]. Generally, the selection of extraction solvent should
limitations. Accordingly, fewer studies related to first-generation bio- follow the premise of “similar dissolves similar”. Case studies between
fuels have been published in recent years, and very little information polarity and extractable compounds mediated by the three commonly
could be found in recent articles regarding the mechanical extraction used organic solvents hexane, chloroform, and acetone are described in
methods applied in first-generation biofuels. For example, among the Table 3. The compositions of the biofuels extracted by different solvents
recent publications only one article was published concerning the from same feedstock are normally dependent upon the polarities of the
synthesis of biodiesel from sunflower-extracted oil [42]. Therefore, solvent used.
considering the sustainability factor, successive generations of feed- As a PSSE method, ultrasonic associated extraction was used for
stocks are extensively analyzed in the current review with a particular liquid biofuel production from Jatropha seeds. This method potentially
focus on extraction in third-generation biofuel feedstocks. reduces the recalcitrance of lignocellulosic materials (lignin, cellulose,
and hemicelluloses) and improves their suitability as bioethanol feed-
stocks [45]. Alternatively, the application of SFE to lignocellulosic
3.2. Extraction methods for the feedstocks in second-generation biofuels biomasses was considered to be appropriate in terms of the high re-
solvability of this method [33]. The majority of studies regarding SFE
Second-generation biofuels comprise fuels that can be manufactured for biofuels from lignocellulosic biomass have been carried out using
from various types of biomass, which are generally obtained from ve- either ethanol or methanol. Notably, the second-generation biofuels
getal materials but can also include non-vegetal materials. Mechanical obtained from biomasses using SFE are significantly affected by ex-
methods supported by chemical CSE are usually applied in first-gen- traction parameters such as biomass/solvent ratio, extraction tem-
eration biofuel production, which can effortlessly extract vegetable oils perature, extraction time, and operating pressure [33]. Moreover, as a
from arable crops as shown in Fig. 3. In comparison, second-generation novel technique, a highly cost-efficient and environmentally friendly
biofuels are produced from lignocellulosic biomass, industrial, civil, method using liquefied gas operated under normal pressure and tem-
and agricultural residues, or wastes from which extraction of the re- perature has been reported with regard to the extraction of bio-crude
quired fueling substances is more difficult. In particular, it is especially fuel from vegetal biomass and industrial waste [26,46].
challenging to obtain the required fuel by means of only extraction
methods from lignocellulosic wood biomasses. Accordingly, the ex-
traction methods frequently mentioned in the recent articles for second- 3.3. Extraction methods for the feedstocks of third-generation biofuels
generation biofuels were instead those used for industrial, civil, and
agricultural residues. In contrast to the food oil crops prevalent in first- The main source of third-generation biofuels is microalgae.
generation feedstocks, the use of extractable woody crop oil for bio- Therefore, third-generation biofuels are also termed algae-biofuels. This
diesel fuel has been reported for second-generation feedstocks (Table 1) is presently regarded as a feasible alternative renewable energy re-
[27]. source for biofuel production, which overcomes the shortcomings of
As shown in Fig. 3, all these classified extraction methods are used first and second-generation biofuels. Algae can provide several different
for the feedstocks in second-generation biofuels, from which diverse types of renewable biofuels including biodiesel, hydrocarbons, and bio-
biofuel products can be produced. Few mechanical methods are used in hydrogen. There are many advantages for producing biofuel from algae
the second generation because compared to the first and third genera- as these organisms can yield 15 to 300-fold more biodiesel than tradi-
tions, the oil content of second generation biomass is generally low tional crops on an area basis [47]. In addition, the harvesting cycle of
leading to a poor recovery rate of oil if only mechanical methods are algae is very short and its growth rate is very high. Moreover, high
applied. Some articles described the derivation of fish oil-based biofuels quality agricultural land is not required for algae biomass production
from fish processing plant waste following application of a mechanical [48]. These undoubted advantages have led algae biofuels to receive
extraction method, whereupon the fish oil was subsequently recovered increasing attention worldwide. Unlike for terrestrial oily crops, me-
using carbon dioxide (SFE) [43] or Soxhlet extraction (CSE) [24]. chanical extraction such as via oil expellers/presses cannot be em-
Different conventional organic solvents have also been tested as ployed for extracting algae oil owing to their small cell size, complex
extraction agents for second generation biofuel production; re- cell membrane, and thick/rigid cell wall [49,50]. Accordingly, various
presentative examples are listed in Table 3. Non- or aprotic polar sol- alternative extraction methods have been and are being tested to ef-
vents hexane, petroleum ether, and chloroform were used for the ex- fectively obtain fueling substances from fresh or pro-treated algae, as
traction of hydrocarbons from Nicotiana glauca (tree tobacco), yielding algae biofuel production is still in the developmental stage (Fig. 3). In
total hydrocarbons of 6.28, 5.69, and 6.20 μg mg−1 FW (fresh weight), contrast to those from second-generation feedstocks, the biofuels ob-
respectively [25]. In another biodiesel production process, rubber seed tained from algae by means of extraction processes primarily comprise
oil was extracted by shaking with non-polar or polar solvents hexane, biodiesel and bio-jet fuel (Table 1), with this production depending on
dichloromethane, or acetone at room temperature for 30 min, resulting the different components biosynthesized by the various species of

Table 3
Case analysis of representative organic solvents applied in second generation feedstocks.
Organic solvent Feedstocks studied Extracted biofuels and yields Ref.
(extractable compounds)

Hexane (Non-polar; hydrocarbons and triacylglycerols) Tree tobacco Hydrocarbons, 6.28 μg mg−1 FW [25]
Rubber seed Bio-oil, 20.4 wt% [27]
Petroleum ether Tree tobacco Hydrocarbons, 5.69 μg mg−1 FW [25]
Orchid tree Bio-oil, 14.8% wt [43]
Chloroform (Aprotic polar; hydrocarbons, triacylglycerols, waxes, etc.) Tree tobacco Hydrocarbons, 6.20 μg mg−1 FW [25]
Dichloromethane Rubber seed Bio-crude oil, 16.0 wt% [27]
Acetone (Polar; phospholipids, glycolipids, diacylglycerols, etc.) Rubber seed Bio-crude oil, 18.0 wt% [27]

FW, fresh weight; wt., weight. The polarity of solvents from hexane to acetone decreases in the order of non-polar to polar.

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Table 4
Case studies of representative extraction methods in algae biofuel production.
Method Species studied Solvents and/or operating conditions Compound, yield Ref.

Soxhlet (CSE) Nannochloropsis sp. Hexane: methanol (3:2), 70 °C, NP Lipid, 30.8% [51]
Bligh-Dyer (CSE) Schizochytrium sp. Methanol-chloroform, 25 °C, NP Lipid, 22.1% [23]
Sonication (PSSE) Chlorella sp. 50 kHz, 15 min, NP Lipid, 25.5% [54]
Microwave (PSSE) Botryococcus sp. 2450 MHz, 100 °C, 5 min, NP Lipid, 28.5% [55]
SFE Tetraselmis sp. SCCO2, 40 °C, 15 Mpa Lipid, 10.8% [57]
Liquefied gas (novel) M. aeruginosa Dimethyl ether, 25 °C, 0.51 Mpa Bio-oil 40.1% [59]
B. braunii Hydrocarbons 48.9% [19]

Yields in dry weight base; NP, normal pressure.

microalgae. Case studies of representative extraction methods for algae Tetraselmis sp. (Table 4). Pilot-scale SCCO2 analyses for the recovery of
biofuel production are listed in Table 4. biofuels from microalgae were reported in 2012 [34]. In a more recent
study, lipid yield and oil composition from Scenedesmus almeriensis ex-
3.3.1. Studies of CSE in algae biofuel production tracted using SCCO2 were comparable to those from CSE via Soxhlet
Table 2 presents representative case analysis of classified extraction extraction and several organic solvents. In particular, results show
methods employed in algae biofuel production. As a widely applied SCCO2 to be the most efficient methodology to extract lipids from algae
standard method, various organic solvents have been tested for algae for further biodiesel production [32]. However, high infrastructure and
biomass extraction. Because different types of lipids comprise the main operational costs associated with SCCO2 remain as primary dis-
components in algae biomasses used for biofuel production, both polar advantages. Alternatively, subcritical water (SuCH2O) has been studied
and non-polar organic solvents have been utilized [30]. Polar organic for generating liquid transportation fuels from algae, with the author
solvents such as methanol disrupt hydrogen bonding between polar claiming that this technology could in principle be used for the pro-
lipids whereas non-polar solvents such as hexane disrupt hydrophobic duction of biofuels and bio-products from algae and other biomasses
interactions between non-polar/neutral lipids. Soxhlet and Bligh-Dyer [35].
[17] extraction constitute the two typically applied methods for the
extraction of lipids from algal biomass (Table 4), which utilize hexane 3.3.4. Studies of novel extraction methods in algae biofuel production
(non-polar), chloroform (aprotic polar), and methanol (polar) as sol- Several novel extraction methods have been recently studied for
vents, respectively. In a recent study, Soxhlet with hexane and me- algae biofuel production. Switchable hydrophilicity solvents were used
thanol (3:2) yielded 30.8 wt% lipids from Nannochloropsis sp. [51]. The for lipid extraction from lyophilized microalgae (Botryococcus braunii)
Bligh-Dyer method with methanol and chloroform (1,1) yielded 22.1 wt [38]. The application of lyophilization can simultaneously dry and
% lipids from Schizochytrium sp. [23]. Numerous other solvents have disrupt algae cells. In addition, ionic liquids mixed with methanol at a
also been tested and mentioned in recent articles, such as butanol and volume ratio of 1:1 was used to dissolve algal biomass, leaving lipids
diethyl ether [52]. The disadvantages of CSE for algae biofuel pro- insoluble [21]. This method allows the lipids to be easily recovered as
duction are that pre-treatment processes such as drying and cell dis- undissolved lipids are lighter than the ionic liquid and methanol mix-
ruption are generally required prior to CSE to remove the water from ture. Ionic liquids were also reported to have the ability to extract
algae cells and release the target components contained within the cells. branched, unsaturated hydrocarbons from the phototrophic microbes
Such processes are highly energy intensive; furthermore, the use of Synechocystis sp. and B. braunii [29]. Moreover, a new publication de-
organic solvents also results in environmental damage. monstrated photocatalysis to be an efficient method for bio-oil extrac-
tion from Nannochloropsis oculata [58]. Notably, liquefied gas as an
3.3.2. Studies of PSSE in algae biofuel production organic solvent can not only extract the biofuel from second-generation
Physical methods are usually utilized to support CSE for the ex- biomasses [26,46] but has also been reported to facilitate the recovery
traction of lipids from algae biomasses, such as application of micro- of bio-oil from wet algae via a shaking extraction method [18]. Using
waving and sonication, bead beating, autoclaving, grinding, or osmotic liquefied dimethyl ether at room temperature (25 °C) and under
shock [53]. Sonication and microwaving are the most frequently used 0.51 Mpa, 91.0 wt% of bio-crude oil and 48.9 wt% of hydrocarbons
PSSEs (Table 4). Recent studies reported that the inclusion of sonication were obtained from Microcystis aeruginosa and B. braunii [19,59].
at 50 kH for 15 min yields 25.5 wt% of lipids from Chlorella sp. [54]
whereas applying microwaves at 2450 MHz and 100 °C yields 28.5 wt% 4. Considerations for selection of extraction methods for algae
lipids from Botryococcus sp. in 5 min [55]. CSE assisted by sonication, biofuel production
bead beating, and heating was also applied for dry algae biomass [28].
A combined method has also been reported that resulted in a higher Fig. 4 depicts the concluding steps of the whole algae biofuel pro-
lipid extraction yield by first heating an algal paste in a continuous duction process for each classified extraction method. Here, the energy
microwave system followed by CSE with hexane [56]. The limitation of demand for algae cultivation, harvesting, concentration, and biofuel
these applied physical methods is that their energy demand is generally refinery are assumed as a fixed constant because these processes are
high; for example, the energy requirement of microwaving and soni- independent from the extraction process in relation to the total energy
cation for 1 kg of biomass is 420 and 132 MJ kg−1, respectively input of algae biofuel production. Conversely, the drying and cell dis-
(Table 2) [56]. ruption processes are treated as variable. The energy requirements
(Units in MJ kg−1 for dried algae biomass) of each upstream process
3.3.3. Studies of SFE in algae biofuel production prior to extraction are assessed based on recently available literature
SFE constitutes a promising green technology method that has the [31,60–62]. For the CSE route, both drying and cell disruption are in-
potential to displace CSE for algae biofuel production [32]. Super- dispensable and have an energy demand that accounts for more than
critical carbon dioxide (SCCO2) represents a good option for SFE as it 80% of the input energy in the upstream process prior to extraction.
offers high solvating power and low toxicity [30]. For example, Li et al. Therefore, the selection of extraction methods for minimizing the use of
[57] reported that at near room temperature (approximately 40 °C) and drying and cell disruption processes is key for reducing the total input
below 15 Mpa the SCCO2 method yields 10.8 wt% lipids from energy demand of algae biofuel production. Alternatively, only the

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Cultivation (65.4 MJ kg-1 ) [60]

Harvesting and concentration (0.64 MJ kg-1 ) [60]

Drying (>25 MJ kg-1 ) [60], (13.8 MJ kg-1 ) [62] Combining drying Free from drying
and cell disruption and cell disruption
(CDC route) (FDC route)

Cell disruption Cell disruption • Such as lyophilization

• Sonication: 132 MJ kg-1 19 MJ kg-1 [61]
may not be needed
[31]
• Microwave: 420 MJ kg-1
[31]

CSE PSSE SFE and Novel Novel

Further processing for the production of biodiesel and bio-jet fuel

Fig. 4. Diagram representing the concluding steps of the algae-to-biofuel flowchart for the classified extraction methods. The processes within the solid outline are
assumed as a fixed constant and within the dashed outline are assumed as a variable. The energy requirement units in MJ kg−1 for dried algae biomass are assessed
based on recent available literature.

drying process is needed for most PSSE methods studied in recent ar- products algal consortium project (NAABB) indicated that an emerging
ticles because physical and/or chemical cell disruption methods were extraction technology (wet solvent extraction) may be selectable to
integrated into the extraction process. However, as the representative provide scale-up data and have sufficient capabilities to produce lipids
physical cell disruption methods, microwaving and sonication impose a for the NAABB program [66]. Nevertheless, the assessment of input
high energy burden and have a high maintenance cost [63]. energy demands for these possible methods is a large and complex task
The representative methods of SFE for algae biofuel production are owing to the differences in the physical and chemical properties of each
SCCO2 and SuCH2O, both of which use lyophilization as a pre-treatment applied solvent and method. Therefore, the logical outcome of the
method. The advantage of lyophilization methods is that they combine present review is that under the current scenario, a potential extraction
both drying and cell disruption in a single process [64] and are there- method should minimize the use of pre-treatment processes to the
fore termed the combining drying and cell disruption (CDC) route. A greatest extent possible.
recently developed switchable solvent extraction method also used
lyophilization as a pre-treatment method [38]. In an experimental 5. Summary
small-scale model, microalgae were pre-treated by lyophilization with
the reported energy demand being 19 MJ for 1 kg of dried algal biomass This review was intended to fill the gap created by the lack of
[61]. Considering that in the case of a large-scale model the energy current review papers discussing recently published academic articles
demand of lyophilization should be lower than 19 MJ kg−1, it might be concerning studies of extraction processes in biofuel production. To
expected that the CDC route would reduce the energy burden in com- clarify the multiple existing and emerging extraction techniques, pre-
parison with CSE and PSSE routes. However, SFE itself may be more dominantly utilized physical and chemical methods were classified as
energy-intensive than CSE and PSSE owing to the use of high pressure CSE, PSSE, SFE, and novel methods. The relationship of feedstocks of
and temperature (Table 2) although it offers advantages such as high different generation biofuels with particular classified extraction tech-
selectivity. niques was confirmed. The extraction techniques already applied in the
Alternatively, some recently developed methods have been reported first-generation biofuel industry mainly relied on the mechanical
that could be free from drying and cell disruption pre-treatment pro- method supported by CSE [67] whereas those for second and third-
cesses (known as the FDC route). An example of such a route is the use generation biofuels are still under development, although most of the
of liquefied dimethyl ether, which serves as a novel solvent that can classified techniques might be employed if the energy demand could be
directly extract lipids and hydrocarbons from undamaged wet algae ignored. Theoretically, SFE has numerous advantages over CSE and
cells [18,19]. Numerous possible extraction methods are currently PSSE including the elimination of organic solvents; i.e., reducing the
being studied based on various ideas with several solvents having been risk of storage, along with rapid extraction speed. However, to date SFE
tested for these processes [65,66]. In an updated study of high-quality is still predominantly applied only in the food and pharmaceutical in-
aviation biofuel production, crude algae lipids were extracted com- dustries. At its current stage, SFE retains a technical bottleneck for large
parably from wet or dry Nannochloropsis with a ratio of algae:hex- scale application in the biofuel production field owing to its inefficient
ane:ethyl alcohol = 1:6.7:3.3 in association with ultrasound [65]; the cost.
final biofuel yield from dry and wet algae lipids was 73.02% and As a new generation biofuel source, the extraction techniques used
76.47%, respectively. Another study on an extraction program within and/or studied in algae biofuel production were intensively reviewed.
the recent 3 year National Alliance for Advanced Biofuel and Bio- Currently, various methods, ideas, and theories regarding extraction

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