Green Chemistry Letters and Reviews

ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/tgcl20

The advances and limitations in biodiesel

production: feedstocks, oil extraction methods,
production, and environmental life cycle
assessment

Konstantin Pikula , Alexander Zakharenko , Antonios Stratidakis , Mayya

Razgonova , Alexander Nosyrev , Yaroslav Mezhuev , Aristidis Tsatsakis &
Kirill Golokhvast

To cite this article: Konstantin Pikula , Alexander Zakharenko , Antonios Stratidakis , Mayya
Razgonova , Alexander Nosyrev , Yaroslav Mezhuev , Aristidis Tsatsakis & Kirill Golokhvast
(2020) The advances and limitations in biodiesel production: feedstocks, oil extraction methods,
production, and environmental life cycle assessment, Green Chemistry Letters and Reviews, 13:4,
11-30, DOI: 10.1080/17518253.2020.1829099

To link to this article: https://doi.org/10.1080/17518253.2020.1829099

© 2020 The Author(s). Published by Informa Published online: 06 Oct 2020.

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GREEN CHEMISTRY LETTERS AND REVIEWS
2020, VOL. 13, NO. 4, 11–30
https://doi.org/10.1080/17518253.2020.1829099

REVIEW

The advances and limitations in biodiesel production: feedstocks, oil extraction

methods, production, and environmental life cycle assessment
Konstantin Pikula a,b, Alexander Zakharenko a,b, Antonios Stratidakisc, Mayya Razgonova a,b
,
Alexander Nosyrevd, Yaroslav Mezhuev e, Aristidis Tsatsakis d,f and Kirill Golokhvast a,b,g
a
Education and Scientific Center of Nanotechnology, Far Eastern Federal University, Vladivostok, Russian Federation; bN.I. Vavilov All-Russian
Institute of Plant Genetic Resources, Saint-Petersburg, Russian Federation; cInstitute for Advanced Study (IUSS), Environmental Health
Engineering, Pavia, Italy; dCentral Chemical Laboratory of Toxicology, I.M. Sechenov First Moscow State Medical University, Moscow, Russian
Federation; eDmitry Mendeleev University of Chemical Technology of Russia, Moscow, Russian Federation; fLaboratory of Toxicology, School
of Medicine, University of Crete, Heraklion, Greece; gPacific Geographical Institute FEB RAS, Vladivostok, Russian Federation

ABSTRACT ARTICLE HISTORY

Although fossil fuels remain the main source of energy, the volume of renewable sources of energy Received 21 December 2018
is constantly increasing. Biodiesel is a promising alternative fuel due to the number of advantages Accepted 19 September
compared to fossil fuel and other types of biofuel. The specific objective of this study was to 2020
identify the difference between conventional and novel technologies applied during the whole
KEYWORDS
life cycle of biodiesel production and consumption. The study offers some important insights Biodiesel feedstock; fatty
into the recent advances in the biodiesel industry including biodiesel production from acid methyl esters;
microalgal lipids, advanced homogenous and enzymatic transesterification, non-catalytic vegetable-oils; oil extraction
supercritical transesterification, application of microwave and ultrasound assisting technologies. methods; environmental life
Considering all the factors affecting the efficiency and safety of the biodiesel production cycle assessment
process, here we reviewed the main principals and recent achievements in the environmental
life cycle assessment of biodiesel production and consumption.

1. Introduction
TG) decomposition by substitution of glycerol base
Biodiesel is a promising alternative fuel due to the with ethanol or methanol (1). In 1977 Brazilian scientist
number of advantages compared to fossil fuel and Expedito Parente designed the first industrial biodiesel
other types of biofuel (bioethanol, biogas, biomass, production line. Then, in 1989 in Australia was opened
etc.). American Society for Testing and Materials the first plant to produce biodiesel from rapeseed oil.
(ASTM) defines biodiesel as monoalkyl esters produced In 2001 a sudden rise in oil prices stimulated biodiesel
from lipid feedstocks, such as vegetable oils or animal entrance to the world fuel market. According to the
fat. In 1937 Belgian scientist George Chavanne patented U.S. Energy Information Administration (EIA) Monthly
the “Procedure for the vegetable oil transformation for Biodiesel Production Report (March 2020), USA
their uses as fuels” which described the transesterifica- biomass-based diesel production in 2019 was 1.725
tion method for triacylglycerol (TAG, or triglyceride, billion gallons (6.530 billion liters). The official EU

CONTACT Konstantin Pikula [email protected] FEFU8 Sukhanova St.Vladivostok 690090 Russiaa; Aristidis Tsatsakis [email protected] Lab-
oratory of Toxicology, Medical School, University of Crete Voutes, Heraklion 71003 Greece
© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
12 K. PIKULA ET AL.

report Biofuels Annual 2019 declares the production of biofuels are made from edible biomass (15). According
14.170 billion liters of biodiesel by EU countries. to the Biofuels Data Sources provided by the United
Oil and gas depletion and environmental law enforce- States Department of Agriculture soybean oil is the
ment for sulfur oxides, carbon oxides, and greenhouse main raw material in the USA, Brazil and Argentina, rape-
gas (GHG) emissions stimulate the increase of environmen- seed oil is the main biodiesel feedstock used in the EU
tally friendly, renewable, and cost-effective biofuel pro- countries, while palm oil mostly used for biodiesel pro-
duction (2). Unburned hydrocarbons (UHC) and GHG duction in Indonesia, Malaysia, and Thailand. Neverthe-
emission reduction, feedstock availability, relatively less, some disadvantages, such as high cost of raw
simple production, cost-effectiveness, high combustion materials, dependence on food prices, fertile land expro-
efficiency, biodegradability, and low toxicity are some of priation, deforestation, and sensitivity of edible plants to
the main biodiesel advantages (3). Biodiesel combustion environmental conditions made biodiesel production
associated with lower emission of GHG, UHC, carbon mon- from edible plants ultimately ineffective (13).
oxide (CO), and particulate matter (PM) emission com-
pared to conventional diesel fuel (4, 5). It was reported
2.2. Second-generation feedstocks
that more than 90% of biodiesel spill volume can
degrade in the environment for 21 days (6). Besides, bio- A promising alternative of using edible plants for biodie-
diesel presence in oil-contaminated sites can have a syner- sel production is the second-generation feedstocks. Non-
getic effect on improving pollution biodegradability (7). edible plants have many advantages as a source for bio-
The purpose of this paper was to review recent diesel production, such as (1) the ability to grow on con-
research in the field of (1) biodiesel feedstock selection, taminated and unsuitable for agriculture areas with
(2) oil extraction methods, (3) biodiesel production tech- minimal requirements for fertility and humidity, (2) stab-
nologies, and (4) life cycle assessment of biodiesel pro- ility over a wide range of climatic conditions, (3) low
duction and consumption. environmental stress and the same quality of the final
product compared to edible feedstocks, (4) less land allo-
cation and the ability to grow in a mixture with different
2. Feedstocks
plants, (5) potential rehabilitation of degraded lands, (6)
The main step for biodiesel production is the selection of no competition with agricultural food sources, (7) resist-
feedstock. A known fact that each type of feedstocks has ance to pests and diseases, (8) the possibility to
a different composition of fatty acids, which defines the produce useful by-products, (9) availability, renewability,
properties of biodiesel, and finally it affects the life-cycle biodegradability, low sulfur, and aromatic hydrocarbons
of biodiesel (8, 9). Physical and chemical properties of oils content (16). Among the main disadvantages of second-
obtained from different feedstocks described in many generation feedstocks are lower oil yield and higher
reviews (8, 10, 11). Depending on the raw feedstock, biodie- alcohol consumption during the production process com-
sel can be classified as one of four biofuel generations: (1) pared to edible sources (10). The diversity of non-edible
produced from edible oil, (2) produced from oils of non- plants with relatively high oil content and their appli-
edible plants, (3) produced from microalgae lipids, used cation as biodiesel feedstock was demonstrated in
cooking oil, or waste animal fats, and (4) produced from many published studies (17–21). The ability of non-
genetically modified microorganisms (12). Given classifi- edible plants to grow in different climatic conditions
cation includes only the origin of biodiesel but not con- allow creating a stable production of biofuels for many
siders the quality and properties of the final product countries. In Table 1, we listed several common and
obtained from diverse feedstocks. The section below promising second-generation biodiesel plants.
briefly describes current state, development, application,
advantages, and limitations for each generation of biofuel.
2.3. Third-generation feedstocks
Third generation biodiesel feedstocks include used
2.1. First-generation feedstocks
cooking oils (UCOs), waste animal fats (WAFs), and
The production of first-generation biodiesel from edible microalgal lipids.
feedstocks was quite popular at the beginning of the
biodiesel era because of raw material availability and 2.3.1. Used cooking oil and waste animal fat
relatively simple biodiesel production process (13). The world population’s growth causes an increase in the
Rapeseed, canola, soybeans, castor, coconut, palm, volume of food industry wastes and by-products (31).
sunflower, and corn are common conventional sources Biodiesel production from UCOs and WAFs becomes a
of oil for biodiesel (14). Up to now, over 90% of total good opportunity for their effective utilization with
 GREEN CHEMISTRY LETTERS AND REVIEWS 13

Table 1. Non-edible plants used for biodiesel production.

Countries and Oil content, %
Source Environmental requirements regions wt. Productivity and features Ref.
Cottonseed tropical and subtropical regions China, India, USA, 17–25 has non-glyceride residuals, such as (10,
Brazill, Pakistan sterols, carbohydrates, gossypol, resins, 22)
phospholipids, and pigments
Jatropha semi-arid areas, marginal areas, can grow on Mexico, India, Brazil, 20–60 the highest productivity after 5 years of (23)
drought stress and nutrient-poor soil Argentina the plantation; can be collected two
times in a year
Jojoba can survive on arid areas, wastelands, and deserts USA, Israel, Mexico, 48–56 presence of complicated wax esters (24)
India, South Africa causes inconvenience in biodiesel
production
Karanja can survive in heat, drought, salinity and frost India, South-Eastern 25–40 the highest productivity after 4–7 years of (10,
conditions Asia, Australia plantation 25)
Linseed semi-arid areas, relatively cool temperatures, Canada, China, 33–47 source of high-quality proteins, soluble (26)
fine-textured and loamy soils, sensitive to the Kazakhstan, India fibers, and polyunsaturated fatty acids
amount of rainfall and moisture conditions
Mahua arid areas, different soil conditions Southern Asia 35–50 the highest productivity after 10 years of (10,
plantation. 27)
Neem diverse climatic conditions (except long-lasting India, Argentina, 20–30 the highest productivity after 10–15 year (10,
cold and freezing), different types of sand, China, Brazil, of plantation 28)
almost all types of soil Egypt
Rubber rainforest areas with high humidity and Thailand, Indonesia, 42–51 (shell), the highest productivity after 6–9 year of (10,
seed precipitation, sensitive to salinity Vietnam, China. 50–60 plantation 29)
India (kernel)
Tobacco tropical climate and woodlands areas China, Brazil, India, 40–50 grows only for one season but can be (30)
USA, Argentina harvested multiple times per year

additional environmental, economic, and food security as the best candidates for this purpose (40). Cultivation
advantages (32). Among the main limiting factors of of microalgae attracts increasing attention as a source
industrial application of oil and fat wastes are the impur- of lipid for biodiesel (41). Using of microalgae as a
ity and the lack of a centralized system for collecting source of lipids has many advantages (42, 43), such as
such row material (32). (1) simple cultivation, (2) high rate of biomass growth
UCOs have a high concentration of free fatty acids with high lipid content, (3) no need to alienate fertile
(FFAs), which adversely affect the yield of biodiesel. soil, and (4) no dependence on the cost of food
According to the content of FFAs, UCOs are divided resources. A complete life cycle of microalgae ends
into two groups: (1) yellow grease (FFA < 15%) and (2) within a few days and the cells reproduce themselves
brown grease (FFA > 15% and water). Yellow grease is using sunlight as a source of energy and carbon
a common raw material for biodiesel production and it dioxide as a source of carbon (44). Moreover, the
can be used after filtration and cleaning (33, 34). biomass growth rate can be increased with the addition
Animal fats are generally formed of a mixture of TGs, of specific nutrients and the optimization of aeration
proteins, water, and diverse minerals (35). WAFs often (45). Microalgae can be cultivated either in open
collected from animal slaughterhouses or fish factories (environmental) or closed (photobioreactor) systems.
(36). Biodiesel derived from WAFs has a high cetane Closed systems commonly used in large-scale biodiesel
number but low pour point due to the presence of production because of the ability to provide lower con-
FFAs and low oxidative stability due to the lack of tamination and controllable cultivation conditions (light
natural antioxidants (37). There are many published illumination, the CO2 feed, the cultivation medium feed,
studies (35, 38, 39) that describe feasibility and features and the circulator system) (46).
of biodiesel production from such sources as tallow Since microalgae grow in a water column, the
(cattle, ship, and goat), lard (pig), poultry fat (birds and biomass content per unit of occupied area is significantly
fish). According to the U.S. Energy Information Adminis- higher in comparison with other plant sources (13).
tration (EIA) Monthly Biodiesel Production Report, EU Among the other positive environmental effects of
countries, USA, China, Brazil, and Canada are rhe main using microalgae are: (1) bio fixation and removal of
produsers of biofuel from UCOs and WAFs. carbon dioxide from industrial flue gases (47), (2) remov-
ing of carbon, nitrogen, phosphorus and other micronu-
2.3.2. Microalgae trients from wastewaters (48), (3) possible utilization of
Microorganisms e.g. bacteria, yeasts, cyanobacteria, and the biomass remained after oil extraction as a source
microalgae can be used in the field of biofuel pro- of ethanol or methane production, as a nutrient in
duction. Cyanobacteria and microalgae are considered animal husbandry, or to be used as organic fertilizer in
14 K. PIKULA ET AL.

agriculture (49). Besides, microalgae biomass contain by- the environment, removal of nutrients from the ecosys-
products useful for pharmaceuticals, food production, tem reducing the biodiversity of the flora and fauna,
fisheries, cosmetics, and other areas (50). changing aquatic ecosystems, and other (66). Although
However, the high amount of unsaturated lipids the fourth generation of biodiesel is under the early
made biodiesel produced from microalgae biomass stage of development, in the future it could overcome
less stable. Other drawbacks of microalgae as a biodiesel the disadvantages of the first three generations and
feedstock are the need for a high amount of nutrients become the most effective replacement of fossil fuel (69).
(nitrogen and phosphorus) (51) and the ability of
certain species to produce harmful toxins (52). Despite
3. Oil extraction methods
this, many researchers consider microalgae as the only
renewable energy source that could completely Oil extraction techniques are an important step in bio-
replace fossil fuels (53). Moreover, microalgae biomass diesel production. There are three main methods of oil
can be processed into a broad spectrum of promising extraction from organic materials: (1) mechanical extrac-
biofuels, such as gasoline fuel, jet fuel, diesel fuel, tion (70, 71), (2) solvent extraction (72, 73), and (3) enzy-
heavy oil, or green diesel (54, 55). matic extraction (74, 75). The choosing of the oil
The content of fatty acids varies for different microal- extraction technique strongly depends on the feedstock
gae species and can have a significant effect on biodiesel source and forms a significant part of the efficiency and
quality. The content of fatty acids also depends on the cost of biodiesel production (76).
cultivation conditions and growth phase of microalgae Mechanical extraction is the most inexpensive method,
culture (56, 57). The essential parameters of choosing however, extracted oil required further filtration and
microalgae species for biodiesel production are (1) phospholipid removal (77). Moreover, presses used for
high growth rate (58), (2) content of FFA and TGs (57, mechanical oil extraction mostly specialized for specific
59), (3) sensitivity to environmental conditions (tempera- plants and ineffective for a variety of feedstocks. It was
ture, pH, salinity, nutrient intake, illumination) (58), (4) assumed that solvent extraction can be cost-effective
sensitivity to carbon dioxide content and aeration (45), for biodiesel production from 50 tons per day and
resistance to other microalgae species and/or pathogens higher (76). Extraction with n-hexane is the most widely
(bacteria, fungi, viruses) (60), (5) simple biomass separ- used technology. However, the application of this
ation and further processing (61), and (6) the possibility method is associated with the risks of a negative impact
of obtaining additional valuable components (50). Many on the environment and human health through the pro-
studies compared different microalgae species and duction of sewage and emissions of volatile organic com-
suggested useful models of biodiesel production from pounds (70). The practice of using mixtures of polar and
algal lipids (62–65). nonpolar solvents is used for oil extraction to ensure com-
plete recovery of neutral lipids. The most frequently used
mixtures are hexane/isopropanol or chloroform/metha-
2.4. Fourth generation
nol with the addition of water (78).
If the species isolated from nature do not suit the indus- This section aimed to review state-of-the-art achieve-
trial requirements, a promising direction is the application ments provided stable and effective methods of oil
of genetic and metabolic engineering to improve the extraction, such as accelerated solvent extraction, micro-
characteristics of oil-producing microorganisms (66). wave-assisted extraction, ultrasound-assisted extraction,
Microalgal modification is mainly focused on lipid and supercritical fluid extraction, and enzymatic extraction.
carbohydrate metabolism, improved nutrient use
efficiency, hydrogen production, improved photosyn-
3.1. Advanced solvent extraction techniques
thesis efficiency, higher stress tolerance, enhanced cell
disintegration, and flocculation (67). Moreover, genetic 3.1.1. Accelerated solvent extraction
modification can contribute oil extraction from microalgal Accelerated solvent extraction (ASE) or pressurized
biomass by inducing autolysis and product secretory solvent extraction is an advanced chemical extraction
systems (40) The most applied genome editing tools method. The essence of the method lies in using sol-
are: zinc-finger nuclease (ZFN), transcription activator- vents at elevated pressure and temperature. The extrac-
like effector nucleases (TALEN), and clustered regularly tion rate increases with increasing temperature, while
interspaced palindromic sequences (CRISPR/Cas9) (68). high pressure prevents the solvent from boiling at
Despite significant advantages, genetically modified higher temperatures. As a result, the reaction time and
species represent a serious environmental and human solvent consumption are significantly reduced com-
health risk, including releasing toxic algae strains into pared to other solvent extraction methods (79).
 GREEN CHEMISTRY LETTERS AND REVIEWS 15

The previous study showed that ASE was the most required preliminary damaging of the cells to gain the
effective and faster method for lipid extraction com- oil (96). Several methods have been developed for
pared to sonication, Soxhlet extraction, and homogeniz- microalgae cell disruption, such as acid addition, soni-
ation (80, 81). Moreover, used solvent or solvent systems cation, autoclaving, and bead-beating (97, 98). MAE
significantly influence the efficiency of ASE. The choos- was reported as the most effective and easiest way to
ing of the solvent system should depend on the simultaneous cell disruption and lipid extraction from
content of lipid fractions (neutral, FFA, and polar). microalgae biomass (94). Moreover, MAE provides an
Several reports demonstrated that chloroform:methanol opportunity to use water as a solvent reducing negative
(2:1) or dichloromethane:methanol (2:1) were the best influence on the environment and allowing oil extrac-
solvent systems for lipid extraction using ASE (80). Cur- tion from wet microalgae biomass (99, 100). The main
rently, many studies have confirmed the effectiveness factors determining the efficiency of MAE extraction
of the ASE method for lipid extraction from microalgae include used type and quality of the feedstock, power
biomass (82–84). of the microwave, volume and properties of the used
solvent, and temperature (101, 102).
3.1.2. Ultrasound-assisted extraction A comprehensive literature review described the
Ultrasound-assisted extraction (UAE) has been con- efficiency of MAE used with different microalgae
sidered as a key technology in achieving the objective species under different working conditions including
of sustainable “green” chemistry (85). The principle of time and power of microwave treatment, type and
UAE consists of the formation of acoustic waves in a volume of the used solvent system, and the ratio of
liquid medium by ultrasound exposure and creating solvent to biomass (100).
cycles of high and low pressure. During the high-
pressure cycle, small vacuum bubbles formed in the 3.1.4. Supercritical fluid extraction
low-pressure cycle are destroyed intensively, creating a Supercritical fluid extraction (SFE) has many advan-
phenomenon called cavitation. High pressure and inten- tages in comparison with solvent extraction, includ-
sive jets of liquid formed during the cavitation mechani- ing high reaction rate, the absence of toxic
cally destroy the cellular structure of the substrate and solvents, the ability to adjust solvent power to accu-
improves mass transfer, facilitating lipid extraction (86). rate extraction of target components, and pro-
UAE was reported as an efficient method of lipid duction of pure crude lipids, which are not
recovery from oleaginous microorganisms, such as contaminated with solvents (103). The most
yeast, fungus, and microalgae (87–89). The authors common supercritical solvent is carbon dioxide
demonstrated the same level of conventional chloro- (104, 105) but acetone, methanol, ethanol, and
form–methanol (2:1) extraction with and without appli- water also used for lipid extraction under supercriti-
cation of ultrasound but ultrasonication reduced the cal conditions (103, 106). Supercritical carbon-dioxide
duration of the process from 8–12 h to 15–20 min (87, efficiently solubilize non-polar compounds while the
89). Moreover, ultrasonication often used as a method extraction polar compounds can be increased using
of simultaneous oil extraction and production of biodie- a polar co-solvent as ethanol, methanol, and acetyl
sel from a variety of feedstocks (90, 91). chloride (107) or azeotropic mixtures as hexane/
ethanol (108). The parameters of SFE, such as extrac-
3.1.3. Microwave-assisted extraction tion temperature, extraction time, biomass/solvent
Microwave-assisted extraction (MAE) is an extraction ratio, and pressure significantly influence the
technology that combines microwave exposure with quality of extracted oil and obtained biodiesel (109,
traditional chemical extraction (92). Microwave electro- 110). Mild critical temperature and the oxygen-free
magnetic radiation is capable of penetrating biomater- operating system allow the successful extraction of
ials, interact with polar molecules such as water, and thermally sensitive lipid fractions without their
eventually heat the entire sample. The advantages of degradation and oxidation (111).
the method are low cost, fast process, low solvent con- The main limitations of SFE are the high cost of the
sumption, high oil recovery, high product quality, process, the need for specialized equipment, the need
reduction of energy consumption, and reduction of for preliminary drying of raw materials to reduce moist-
CO2 emissions. ure below 20% (112). Application of SFE to microalgal
In practice, MAE often used to extract oils from the biomass required substrate pre-treatment (e.g. high
biomass of microalgae (93, 94) or fish waste (95). pressure, bead-beating, ultrasound, microwave, and
Effective extraction of microalgae lipids is complicated other) to enhance mass transfer and facilitate solvent
by the existence of a thick cell wall of microalgae and access to cell content (97, 98).
16 K. PIKULA ET AL.

3.2. Enzymatic extraction The main drawbacks of AEE are the duration, high
cost of enzymes, and additional stage of demulsification
Aqueous enzymatic extraction (AEE) is a promising tech-
(77, 123). Nevertheless, AEE has great prospects and can
nology for extracting oils from plant materials (113). The
become a more affordable and cheaper method in the
advantages of the method include environmental safety,
future.
absence of volatile organic component formation,
reduction in solvent consumption, the possibility of sim-
ultaneous protein and oil extraction without loss of 4. Transesterification technologies
quality, no need to remove phospholipids (114). The
efficiency of oil extraction depends on enzyme concen- There are four basic methods of biodiesel production,
tration, type, and composition of enzymes, type of feed- such as (1) straight using and mixing of raw vegetable
stock, and reaction conditions. oils (124, 125), (2) micro emulsification (126), (3)
The increase in the concentration of enzymes leads to thermal cracking (pyrolysis) (127), and (4) transesterifica-
an increase in the interaction with the substrate, which tion (128).
promotes peptide bonds’ destruction. When the concen- In this section, we will focus only on transesterifica-
tration of enzymes exceeds the saturation point of the tion methods of biodiesel production as it is the most
substrate-active sites, odors, bitterness, and carameliza- used and promising technology. Transesterification is
tion of sugars is possible, which hinders the extraction the process of esters and glycerol formation by reaction
process and affects the quality of extracted oil (115). of triglycerides with low molecular weight alcohols
The presence of water is necessary for diffusion, (methanol or ethanol). In contrast to the other
enzyme mobility, and hydrolytic reactions (116). Low methods, transesterification produces little or no
moisture content leads to the formation of a thick sus- carbon deposition, and the product has properties com-
pension and inhibits enzyme activity (117). At the same parable with petroleum diesel (129). Transesterification
time, high humidity can reduce the concentration of can be divided into (1) catalytic and (2) non-catalytic
enzymes, ergo decrease the extraction grade (118). (130). Catalytic transesterification is classified according
Cellulase, hemicellulase, phospholipase, pectinase, and to the type of used catalyst. Non-catalytic transesterifica-
protease enzymes are often used for oil extraction (119). tion includes reactions under supercritical conditions
Some studies showed that using of enzyme mixtures (131, 132). In practice, biodiesel production is often
was more effective compared to individual enzymes carried out by combined methods (130).
(120), and hydrolysis of protease proteins in protein-rich
plants promotes the release of oils and increases the 4.1. Catalytic transesterification
efficiency of extraction (121). Moreover, the cell wall of
plant materials is composed of cellulose, hemicellulose, A simplified scheme of catalytic transesterification
and pectin all of which can easily be broken down by shown in Figure 1. The release of fatty esters occurs at
the wide range of commercially available enzymes (122). each step of triglyceride conversion to diglyceride,
It was shown that extraction intensity increases with a monoglyceride, and then to glycerin (133). There are
reduction of substrate particle size. The extraction several types of catalysts, such as (1) acidic (homo-
efficiency also depends strongly on the pH of the geneous or heterogeneous), (2) alkaline (homogeneous
medium. Each enzyme has a specific pH optimum and or heterogeneous), (3) enzymatic (134). The selection
the extraction should be carried out away from the iso- of catalyst used for biodiesel production through trans-
electric point at which proteins are insoluble and discou- esterification reaction is determined by the quantity of
rage the extraction process. The next crucial factor is FFAs and the nature of the feedstock (10). The advan-
temperature. The maximum enzyme activity is observed tages and disadvantages of different catalysts are pre-
at the temperature of about 30–45°C, and it varies for sented in Table 2.
different raw materials (114). Attention also should be
paid to the mixing regime, so it is necessary to select
such parameters at which the maximum interaction of
enzymes with the substrate can be achieved, but a
stable emulsion is not formed, facilitating the formation
of the easily detachable oil layer on the surface. More-
over, stirring leads to energy consumption and raising
the cost of the process (122). In practice, AEE is also
Figure 1. Catalytic transesterification scheme. R1, R2, R3 –
used in combination with sonication (123).
hydrocarbon radicals.
 GREEN CHEMISTRY LETTERS AND REVIEWS 17

Table 2. Advantages and disadvantages of transesterification with different types of catalysts.

Catalysts Advantages Disadvantages
Homogeneous alkaline: NaOH, KOH, CH3ONa, availability of catalysts, the possibility of reaction sensitivity to FFAs and water, the need for
CH3OK under atmospheric pressure and temperature, preliminary purification of raw materials (135)
high reaction rate (135)
Heterogeneous alkaline: CaO, MgO, SrO, cheap synthesis of catalysts, the possibility of high reaction temperature, the possibility of by-
Mg6Al2(OH)16CO3.4(H2O) catalyst regeneration and reuse, the activity similar product formation, slower reaction rate (137)
to homogeneous acid catalysts, environmental
safety (136)
Homogeneous acid: H2(SO)4, HCl resistance to FFAs, the possibility of simultaneous high reaction temperature, high alcohol-to-oil ratio,
esterification of FFAs and transesterification of high duration of the process, the ability to promote
triglycerides (138) the corrosion of equipment and materials (138)
Heterogeneous acid: ZrO2, SnO2, TiO2, zeolites, resistance to FFAs, the possibility of simultaneous low activity, high duration of the process, high cost
sulfonic ion-exchange resins, ZrO2/Al2O3/ esterification of FFA and transesterification of of catalysts, undesirable side reactions (139)
3+
WO3, SiO2/ZrO2, SO2−
4 /TiO2/La, ionic liquids triglycerides, low toxicity, lower corrosion attack
than a homogeneous acid catalyst, the possibility
of regeneration and reuse of catalyst (126)
Enzymatic: bacterial, fungal, and yeast lipases resistance to FFAs, the possibility of simultaneous the high cost of enzyme production, low reaction
esterification of FFA and transesterification of rate compared to alkali catalyzed
triglycerides, no need to recover unreacted transesterification, inhibition of enzymes by
methanol and wastewater treatment, low alcohol- alcohol, gradual deactivation and limited
to-oil ratio, simplicity of enzyme separation, regeneration of enzymes (140, 141)
possibility reusing, the biodegradability of
enzymes, absence of by-products, moderate
reaction temperature (30–50°C), high selectivity to
the substrate (126, 140)

4.1.1. Alkaline catalysts a tetrahedral intermediate, (3) destruction of this com-

4.1.1.1. Homogeneous alkali catalysts. The prevailing pound and (4) regeneration of the catalyst. This
part of biodiesel is currently produced with an alkaline sequence is repeated two or more times until the for-
catalyst (142). However, the presence of FFAs in the feed- mation of biodiesel and glycerin (143).
stock can lead to soap and water formation in reaction
with an alkaline catalyst (Figure 2). This reaction is unde-
sirable because of the formation of soap which reduces 4.1.1.2. Heterogeneous alkaline catalysts. It is
the volume of produced biodiesel and slows the separ- assumed that transesterification with heterogeneous
ation of esters from glycerol (143). Moreover, water alkaline catalysts can overcome the disadvantages of
from oils or fats inhibits the transesterification by the process with homogeneous catalysts. However, the
hydrolysis reaction. Water can hydrolyze triglycerides reaction of the catalyst with glycerin may lead to the for-
to diglycerides and form more FFA (Figure 3), which, mation of side products (126, 145). Ca and Mg are most
eventually, inhibit the transesterification. The widely used as heterogeneous alkaline catalysts due to
maximum permissible concentration of FFAs for the their poor solubility in methanol and the strongest cata-
use of alkaline catalysts is 0.5–2.5% of dry weight. If feed- lytic activity among alkaline earth metal oxides (146).
stock contains more than 0.5–2.5% of FFA of dry weight, The formation of calcium diglyceride during transester-
a pretreatment stage is required (144). In industrial pro- ification with CaO is shown in Figure 5.
duction, the content of FFA and water in the feedstock The transesterification mechanism with CaO and
must be strictly controlled to guarantee the quality of methanol includes the following steps: (1) the formation
biodiesel. of methylate anion by cleavage of a proton from metha-
The transesterification mechanism with a homo- nol by the main CaO center, (2) attack of the carbon
geneous alkaline catalyst (Figure 4) includes (1) for- atom of triglyceride carbonyl group by the methylate
mation of an alkoxide, (2) nucleophilic attack of anion to form an alkoxy carbonyl intermediate, (3)
alkoxide on the carbonyl group of triglycerides to form

Figure 2. The scheme of FFA interaction with an alkaline

catalyst. Figure 3. The scheme of triglyceride hydrolysis.
18 K. PIKULA ET AL.

Figure 4. The mechanism of transesterification with a homogeneous alkaline catalyst.

rearrangement of the intermediate with the formation of acid transesterification is applicable for processing of
more stable compounds consisting of biodiesel and an cheap feedstocks with a high content of FFAs. It was
anion of diglyceride, (4) attack of the calcium hydroxide noted that the acid hydrolysis reaction is more
cation by the diglyceride anion to form a diglyceride and effective when the content of FFAs in the raw material
CaO (catalyst regeneration). The reaction mechanism is is higher than 1% wt. (126).
repeated two or more times until the formation of bio- The mechanism of homogeneous transesterification
diesel and glycerin (126). with an acid catalyst (Figure 7) involves (1) addition of
It was reported that the surface of CaO strongly a hydrogen ion to the carbonyl group, (2) nucleophilic
adsorbs FFAs, atmospheric carbon dioxide, and water attack of alcohol to form a tetrahedral intermediate, (3)
lowering biodiesel yield and inhibiting catalyst regener- migration of protons, and (4) destruction of the inter-
ation (145). However, these problems can be overcome mediate. This sequence is repeated two or more times
by loading CaO on carrier or support providing better until the formation of biodiesel and glycerin.
availability of the catalytically active site and stability Despite the resistance to a high level of FFAs in the
of the catalyst (147). raw feedstock, the process of homogeneous acid trans-
Thus, heterogeneous alkaline catalysts can be formed esterification did not find industrial application due to
by incorporation of catalysts with active basic functional the slower reaction rate, requirement of high reaction
groups in different support materials such as carbon temperature, the high molar ratio of alcohol to oil,
(148), lignin (149), clays (150), ionic exchange resins difficult separation of the catalyst, serious environmental
(151), basic ionic liquid (152), and basic organic com- and corrosion-related problems (143).
pounds. Moreover, the availability of active sites of
solid catalysts can be increased by the synthesis of nano- 4.1.2.2. Heterogeneous acid catalysts. Biodiesel produ-
particle size catalyst with high surface area (153). cing with heterogeneous acid catalysts is used on an
industrial scale, despite the low activity. The presence
4.1.2. Acid catalysts of various acid sites with different acid strengths facili-
4.1.2.1. Homogeneous acid catalysts. Homogeneous tates the separation of the catalyst from the reaction
acid transesterification (Figure 6) is carried out in the medium, regeneration, and repeated application of cat-
presence of concentrated sulfuric or hydrochloric acid. alyst (154).
Unlike reactions with an alkaline catalyst, the presence Zeolites are most widely used as solid acid catalysts
of FFAs does not adversely affect acid catalysis. In the for the transesterification of oil (142). The advantage of
presence of an acid catalyst, FFAs can react with alcohols using zeolites as heterogeneous acid catalysts is their
to form esters in the esterification reaction. Thus, the natural origin, cheapness, the ability to control acidity
by modifying the aluminosilicate base. However, such
catalysts have low activity due to steric hindrances,
which limit the diffusion of triglycerides into the

Figure 5. Calcium diglyceride formation in transesterification

with calcium oxide. Figure 6. The scheme of FFA esterification with an acid catalyst.
 GREEN CHEMISTRY LETTERS AND REVIEWS 19

Figure 7. The mechanism of transesterification with a homogeneous acid catalyst.

micropores of the catalyst (155). During the reaction, a 4.1.3.1. Sources of enzymes. Lipase enzymes are used
certain amount of organic matter can accumulate on as biological catalysts to promote the hydrolysis of TGs
the surface of the catalyst and cause further inhibition to glycerol and fatty acids. According to their origin,
of the reaction. these enzymes classified as (1) plant, (2) animal, and
It was reported that after the addition of ZrO2 with (3) microbial lipases. Plant lipases are not commercially
sulfuric acid H2SO4 the yield of biodiesel from palm available. Animal lipases are less stable and more
and coconut oil increased from 64.5% and 49.3% to expensive than microbial. Lipases from bacteria (164),
90.3% and 86.3%, respectively (156). The reason for fungi (165), and yeasts (166) described as effective cata-
increasing the volume of produced biodiesel is the lysts for biodiesel production. Among known lipase-pro-
modification of metal oxides surface acidity. The combi- ducing microorganisms, the most commonly used for
nation of ZrO2-Al2O3-WO3 catalysts can increase the the production of biodiesel are Candida sp., Pseudomo-
mechanical strength and acidity of the catalyst (157). nas sp., and Rhizopus sp. (167).
The properties of various solid acid catalysts were Lipases from different sources have a different inter-
clearly described in the article (143). action with the substrate. It leads to a difference in bio-
Ionic liquids are thermally stable, nonvolatile solvent, diesel yield with different substrates (168). For optimal
which can act as a catalyst for biodiesel synthesis. production of biodiesel, the lipases must be able to
Choline chloride, 4-allyl-4-methyl morpholine-4-ium, 1- convert mono-, di- and triglycerides to biodiesel and
ethyl-3-methylimidazolium methyl sulfate, and 4-(3- efficiently esterify FFA.
methyl-1-imidazolio)-1-butane sulfonic acid hydrogen
sulfate were reported as efficient, low toxic, and environ-
ment-friendly catalysts for biodiesel production in com- 4.1.3.2. Reaction conditions. Methanol, ethanol, propa-
bination with microwave irradiation (158–160). The nol, n-butanol, isopropanol, and isobutanol are used
hydrophobic nature of ionic liquids makes it suitable with enzymatic catalytic transesterification. Methanol is
for the low quality of feedstock with a high content of most often used due to its low cost and high reactivity.
FFAs (161). However, it was noted that short-chain alcohols inacti-
vate enzymes more easily than long aliphatic alcohols
(169). To solve this problem, several options have been
4.1.3. Enzyme catalysts proposed, such as (1) step-by-step methanol introduc-
Chemical catalytic transesterification is complicated by tion (170), (2) using of solvents such as hexane, n-
the need for wastewater treatment and the need to heptane, tert-butyl alcohol, 1,4-dioxane, benzene,
purify the glycerin as an added value product. These pro- chloroform (171), and (3) using of alternative acetate
blems can be avoided by using enzymatic transesterifi- acceptors, such as alkyl ethers (methyl acetate, ethyl
cation (162). Due to their unique properties, the acetate) or long-chain alcohols (isopropanol, t-butyl
enzymes act at solid–liquid interfaces and catalyze the alcohol, octanol) (172, 173).
hydrolysis, transesterification, and alcoholysis reactions The addition of water can increase the activity of
(163). The efficiency of enzymatic biodiesel production enzymes but reduce the yield of biodiesel by triglyceride
depends on many factors, such as (1) nature and proper- hydrolysis and formation of FFA and diglycerides. The
ties of enzymes, (2) method of enzyme immobilization, optimum pH for microbial lipases is 7.5–9 (174).
(3) pre-treatment procedures, (4) feedstock type, (5) Lipases of different microorganisms differ in optimal
extraction conditions, (6) transesterification conditions, temperature. Thus, for lipases of mesophilic microorgan-
and (7) type of used bioreactor (164). isms, the optimum temperature is 35–50°C, for
20 K. PIKULA ET AL.

thermophilic microorganisms 60–80°C (175). However, immobilization (126). Whole-cell filamentous fungal
some thermostable lipases can ensure activity up to lipases immobilized within biomass support particles
100°C. in the presence of solvents and detergents considered as advantageous catalysts. In this case, immo-
(176). Enzymatic biodiesel production is performed in bilization happens spontaneously during cell growth
either batch or continuous system. The variety of bio- (177). Several studies reported higher conversion ability
reactor designs described in many research (162, 177– of the whole-cell lipases from Rhizopus oryzae, Rhizopus
179). chinensis, and Mucor circinelloides compared to
Novozym 435 (189–191). Moreover, genomic methods
4.1.3.3. Enzyme immobilization. Immobilization is a were used to produce lipases with required character-
key part of biodiesel production with enzyme catalysts. istics (192, 193). Genetic engineering is a promising area
Immobilization includes physical attachment of for further investigation to manipulate the lipase gene
enzymes to solid support, which can be natural or syn- for higher biodiesel yield in different conditions.
thetic material (180). Compared to free enzymes,
immobilized enzymes have higher stability, accelerated
4.2. Non-catalytic supercritical transesterification
reactivity, prevent undesirable distribution in waste-
water, and can be easily separated and reused (181). Biodiesel production under supercritical conditions is an
The choice of the immobilization technique depends excellent alternative to catalytic transesterification (194).
on the nature of the substrate, enzyme type, and reac- The mechanism of the reaction of triglycerides with
tion conditions (162). Common methods used for methanol under supercritical conditions was first
lipase immobilization include adsorption, covalent described by Kusdian and Saka (195). It was shown
bonding, cross-linking, and entrapment (177). that a reaction in supercritical conditions can overcome
The immobilizing materials should have a porous many problems of chemical and enzymatic catalytic
surface that allows lipase to be loaded within the internal transesterification (196). Non-catalytic production of bio-
structure. The pores should provide access of substrate to diesel in supercritical conditions exceeds the traditional
the active site of enzymes and output of the products methods of reaction in speed and productivity. It was
(182). Various materials such as polymer resins, celite, demonstrated that the share of biodiesel yield under
silica, ceramics, nanoparticles, magnetic particles, micro- supercritical conditions increased with increasing
spheres, cross-linked protein-coated microcrystals, and molar ratio of oil to solvent from 1:1–1:41 (197).
electrospun nanofibers used as lipase carrier (177). In Methanol (198), ethanol (199), propanol (200), butanol
recent years, different nanomaterials became very prom- (201), methyl acetate (202), dimethyl carbonate (203),
ising support for enzyme immobilization. Enzyme nano- dimethyl tert-butyl methyl ether (204) usually used as
carriers classified as nanometal, gold nanoparticles, reaction medium for transesterification under supercriti-
chitosan nanoparticles, nano-silica, zirconia nanoparti- cal conditions. The increase in density causes a decrease
cles, nanodiamond, nanographene, and nanofibers (183). in polarity due to the addition of hydrogen (194). Thus,
According to the nature of immobilized enzymes, non-polar triglycerides are better dissolved in methanol
they divided into (1) extracellular (180), and (2) intra- under supercritical conditions and form a homogeneous
cellular (whole cells) (140). phase. The solubility of triglycerides increases with
Candida antarctica enzymes immobilized on acrylic, increasing temperature and pressure (205). The addition
ethoxylate resin, or various nanoparticles are the most of co-solvent can increase the mutual solubility of trigly-
commonly used lipases among extracellular enzymes cerides and methanol. Hence, biodiesel can be produced
(184). Novozym 435 is the general name for effective com- under more moderate conditions. Propane, carbon
mercial enzymatic catalyst consisting of immobilized dioxide, acetone, ethane, n-butane, n-hexane, n-
Candida antarctica lipase B on macroporous resin material, heptane, and tetrahydrofuran can be used as co-solvents
produced through submerged fermentation of a geneti- (126, 206).
cally modified Aspergillus niger (185). In several research Comparing transesterification with methanol under
works, Novozym-catalyzed transesterification was com- subcritical and supercritical conditions, it was noted
pared with other commercial lipases such as Amberlyst that the distribution of methanol, triglyceride, biodiesel,
15 (186), Lipozyme TL IM (187), Lipozyme TLIM (188), and and glycerin varied with pressure and temperature (207).
other enzymatic catalysts and lipase mixtures (134). Supercritical production of biodiesel was successfully
The technique of whole-cell immobilization con- used in the processing of feedstocks with a high
sidered one of the most promising methods for enzy- content of FFAs (208). It was found that at 350°C, up to
matic production of biodiesel since it does not require 95% of FFAs were converted to biodiesel in supercritical
the separation and purification of enzymes before methanol (198), and the presence of water does not
 GREEN CHEMISTRY LETTERS AND REVIEWS 21

reduce biodiesel production but even increase pro- heating biodiesel production (219). Microwave heating
ductivity by hydrolysis of triglycerides to FFAs (195). was also used with non-catalytic supercritical transester-
Thus, supercritical conditions allows producing biodiesel ification increasing the heating rate and decreasing the
from the raw biomass with high moisture content such energy consumption of the process (220).
as microalgae excluding oil extraction step by simul-
taneous extraction and transesterification of algal 4.3.2. Ultrasound-assisted transesterification
biomass to biodiesel (209–212). Ultrasound-assisted transesterification can significantly
It was noted that the addition of catalysts can lead to increase the mass transfer between immiscible liquids
an increase in the efficiency of biodiesel production in a heterogeneous system (221). As a result, it reduces
under supercritical conditions (213, 214). Most of the het- the ratio of alcohol to oil, the required amount of cata-
erogeneous catalysts are not stable under supercritical lyst, and consumption of energy for the mixing of the
conditions. Zinc and zirconium aluminates supported liquids. Moreover, the collapse of cavitation bubbles pro-
by a metal oxide catalyst are more stable catalysts vides the generation of free radicals during the soni-
under supercritical conditions (126). Since the hetero- cation and enhances the chemical reaction and
geneous catalysts showed instability, it is possible to biodiesel yield (222).
add trace amounts of homogeneous catalysts under Ultrasound can be applied in pulse or continuous
subcritical conditions to enhance biodiesel yield (215). mode. Pulse sonication creates relaxation intervals pro-
Currently, supercritical transesterification is one of the viding good emulsification of reaction mixture without
most promising techniques for large scale biodiesel pro- a significant increase in thermal energy and preventing
duction. However, there are some drawbacks such as the energy loss (223). Ultrasound-assisted transesterification
low energy efficiency of the process, high required with heterogeneous catalysts can cause catalyst deacti-
temperature and pressure, high oil-to-solvent ratio, vation by long term operation under intense mixing con-
possible coking in the reactor, and the other (126). ditions (224). It was found that enzymatic activity
These limitations made supercritical transesterification increases under low frequency and moderate energy
expensive, restrict large scale biodiesel production ultrasound (225).
under supercritical conditions, and required further
study.
5. Life cycle assessment of biodiesel
production and consumption
4.3. Assisting transesterification techniques
Biodiesel was initially proposed as a renewable, stable,
4.3.1. Microwave-assisted transesterification cheap, safe, biodegradable, and environmentally
Microwave irradiation directly heats the fluid instead of friendly alternative to fossil fuel. However, there are
convection and conduction, hence it prevents heat different concerns related to the possible risks of the
loss, uneven heating, and energy losses (216). Micro- growing biodiesel industry (226). The life cycle assess-
wave-assisted transesterification influences the dielec- ment (LCA) approach has been frequently used to evalu-
tric properties of a blend containing a polar and ionic ate and compare the environmental, energy, and
compound of oil and solvents facilitating fast intermole- economic aspects of biodiesel production and consump-
cular blending and quick transformation of oil into bio- tion (227–230).
diesel. Therefore, microwave irradiation reduces The LCA defined as a standardized ISO method aimed
reaction time, minimizes environmental impact, and to detect pollution transfer from one step to another
consumes less than 10% of energy to achieve the one, or from one kind of environmental impact to
same yield of biodiesel compared to the conventional another one (228). This approach considers the entire
heating method (217). However, the large-scale appli- life cycle of a product or so-called “from cradle to
cation of microwave-assisted transesterification compli- grave” principle (231). The systematic evaluation of
cated by the low penetration depth of the microwaves. mass and energy balances in all of the stages of biodiesel
This drawback can be overcome by using of microwave production is an important tool which can show the less
source with higher frequency, higher power reactor, or effective processes of the production chain and indicate
effective substrate stirring (218). Moreover, the the way of improvement of biodiesel production (232).
efficiency of the process varies when microwave LCA of biodiesel produced from first- and second-
irradiation used with different homogeneous or hetero- generation feedstock includes the following stages
geneous catalytic systems (216). Immobilized enzymes (227): (1) agricultural cultivation, (2) transportation, (3)
can be used at temperature 80–100 °C without reducing oil extraction, (4) biodiesel production, and (5) combus-
their activity and stability, and can be used in microwave tion. For the third-generation biodiesels, the agricultural
22 K. PIKULA ET AL.

cultivation stage includes the cultivation of algae in yield per unit area (244). The LCA research works
pounds (228). However, it is difficult to compare the reported that the land required for agricultural systems
results of different LCA due to variety and lack of uni- was 96% higher than for the algal biodiesel production
formity in assessed functional units (233, 234). Moreover, (245). Moreover, microalgae cultivation can be associ-
comparison of biofuels to fossil fuels in terms of LCA is ated with carbon capture from flue gas emissions and
an important issue discussed in previous studies (235, wastewater treatment (52, 246). It was reported that
236). In this section, we will focus on reviewing the 1 kg of algal biodiesel fixes about 0.6 kg of CO2 (247).
main LCA features of different stages of biodiesel The variety of microalgae cultivation pathways compli-
production. cated the LCA. Life cycle optimization methodology
based on the combination of LCA with mathematical
programming is a promising tool for the selection of
5.1. Feedstock stage
the best technology option and process configuration
5.1.1. Agricultural cultivation for the cultivation system of microalgae (248). LCA
The agricultural cultivation stage could further be studies of different microalgae production systems
divided into the substages (227) such as (1) production were summarized in many reviews (228, 248, 249).
of agricultural inputs (e.g. seed, fertilizers, pesticides,
etc.), (2) production of capital goods (e.g. agricultural
5.2. Transportation and oil extraction stages
buildings and machinery), and (3) fieldwork operations
(e.g. land preparation, planting, fertilizing, harvesting, The transportation stage of LCA includes the following
etc.). A lifetime of a plant is an important point in LCA substages (227): (1) transportation of the agricultural
of first- and second-generation biodiesel. For perennial outputs to the oil extraction plant, (2) transportation of
plants, the above-mentioned cultivation substages the extracted oil to the biodiesel production plant, and
may take place periodically. However, in this case, (3) transportation of biodiesel to the consumer. The oil
additional pre-nursery, nursery, and immature planta- extraction stage can be divided into (227) (1) upstream
tion substages should also be considered (237, 238). activities for the production of input materials (e.g.
Moreover, agricultural feedstocks are coupled with chemicals, fuels, electricity, etc.), (2) production of
land occupation and land-use change which can play a capital goods, and (3) oil extraction plant operations
key role in total GHG emission of the biodiesel life (e.g. oil extraction, wastewater treatment, substrate
cycle. The fertilization used to raise the yields of drying, etc.). All the stages of the biodiesel life cycle
energy crops results in increase of nitrous oxide gener- include the risks of accidents, such as equipment
ation, and deforestation decreases CO2 absorption failure, human factor, operator error, external event,
(239). Hence, deforestation, meadow degradation, the ignition, etc. (250). Consequently, it can result in injuries,
tillage system, and locations (air temperature, soil fatalities, partial material loss, oil, or fuel spills, which
organic carbon content) will influence the environ- would have a negative influence on soil or water
mental impacts of the agricultural stage (240). environment (242, 251, 252). The current understanding
of biodiesel fate, transport, and bioremediation in the
5.1.2. Used cooking oil and waste animal fat environment was discussed in several state-of-the-art
supply chain reviews (253, 254).
The environmental safety and stability of biodiesel pro-
duction from UCOs and WAFs are still not well examined
5.3. Biodiesel production stage
(241, 242). Although the higher hazards of biodiesel pro-
duction from UCOs and WAFs were associated with bio- The biodiesel production stage can further be divided
diesel production stage, the material collection and into substages (227), such as (1) production of input
transportation also represent serious risks attributed to materials (e.g. chemicals, electricity, etc.), (2) production
accidents and road transport (242). One of the main of capital goods, and (3) biodiesel production plant
limitations of large-scale biodiesel production from operations (e.g. biodiesel production, biodiesel
UCOs and WAFs is the absence of centralized systems refining, wastewater treatment, etc.). The type of biodie-
of raw material collection from commercial or domestic sel production technology significantly affects the life
sources (243). cycle of the final product. In practice, after the transes-
terification process remains sub-products such as trigly-
5.1.3. Microalgae cultivation cerides, diglycerides, monoglycerides, biodiesel esters,
The main advantage of microalgae as biodiesel feed- glycerol, alcohol, and catalytic components (255). More-
stock is the cultivation conditions and high biomass over, transesterification associated with the
 GREEN CHEMISTRY LETTERS AND REVIEWS 23

consumption of a high amount of energy for heating improve emulsion stability and reduce the ignition
and stirring and using catalysts (256, 257). Despite exist- delay period (268). The air–fuel mixing and complete
ing shortcomings, the use of CO2 in supercritical lipid combustion of biodiesel can be improved by nanoaddi-
extraction and transesterification processes or tives, such as Al, ZnO, TiO2, and Ag (269). Moreover, it
implementation of enzymatic catalysts could signifi- was reported that fuel nanoadditives improves the
cantly reduce the environmental impact of biodiesel soot oxidation by reaction with carbon atoms (270).
production as it was described before (132, 177, 258). The main disadvantage of nanoadditives is the appear-
ance of the nanoparticles in the engine exhaust. The tox-
icity of nanoparticles to the environment and human
5.4. Combustion stage
health have been reported in many research papers
The combustion stage considers emissions from station- (271–273).
ary or mobile engines running on biodiesel. Tradition-
ally, the environmental impact of biodiesel combustion 5.4.2. The influence of engine type and operating
associated with lower emission of GHG, unburned conditions on emission content
hydrocarbons (UHC), carbon monoxide (CO), and par- The exhaust emissions of diesel engines can also be
ticulate matter (PM), but higher nitrogen oxides (NOx) reduced by modifying the geometry of the combustion
emission compared to conventional diesel fuel (4, 259). chamber, modifying the injection timing and com-
However, many recent research works reported lower pression ratio, installing exhaust gas recirculation
NOx emission during the combustion of biodiesel systems, selective catalytic reduction systems, catalytic
under optimized conditions (4, 260). Mostly, the combus- converters, diesel particulate filters, or installing low-
tion emission depends on (1) biodiesel type and quality, temperature combustion technologies (264). It was
(2) engine type, and (3) engine operation conditions noticed that in cold climate regions diesel engines
(261). consume more fuel and emit more CO and UHC (274).
To improve engine performance under cold start con-
5.4.1. The influence of biodiesel quality on ditions, the researchers developed the methods of opti-
emission content mizing the nozzle geometry, formulating fuel blends
The quality of biodiesel depends on many factors, such with improved cold flow properties, application of cold
as the quality of feedstock, fatty acids composition, pro- flow improvers, and preheat systems (264, 275). The
duction, and pre-treatment processes. Industrial pro- diesel particulate filters were applied to reduce PM emis-
duction of biodiesel regulated by the standards ASTM sions of fuel combustion (276). However, the diesel par-
D6751 in the USA and EN 14214 in the EU (262). Many ticulate filters required periodical regeneration, and the
Asian countries have their standards for the quality of optimal materials, structure, and regeneration technol-
biodiesel (263). Moreover, there are various fuel modifi- ogy of the filters are still under discussion (277, 278).
cation techniques such as the inclusion of nanoaddi- Low-temperature combustion technology applied for
tives, alcohols, or water which aimed to obtain the internal combustion engines can significantly reduce
desired engine performance and emission level (260). the overall in-cylinder temperature and facilitate low
The engine performance and exhaust emission par- NOx and PM emissions (279).
ameters of diesel engines can be changed by using
various mixed biodiesel–diesel blends. The changes in
6. Conclusion
the physicochemical properties and exhaust emission
parameters of the blends composed of different feed- The volume of world biodiesel production is constantly
stock biodiesel and diesel fuel were compared in many increasing. Despite the share of biodiesel produced
works (264, 265). The higher oxygen level of biodiesel from edible oils still exceed the share of more promising,
compare to fossil fuel leads to complete combustion of cheap, and stable sources, such as non-edible plant oils
the fuel in a diesel engine and results in lower UHC and microalgae lipids, the prospects of these feedstocks
and CO, and PM emission (266). The addition of the in biodiesel production are obvious. Used cooking oil
limited concentration of water to biodiesel causes a and waste animal fat are the other promising lipid
micro-explosion phenomenon and stimulates a more source for biodiesel production. This raw material will
combustible air–fuel mixture, which was reported as a remain relevant, as it is constantly generated in the
method of the NOx and PM emissions reduction (267). food industry.
However, the water emulsified diesel fuel increases the The efficiency of the biodiesel industry significantly
ignition delay period and decreases engine perform- depends on the achievements of oil extraction from
ance. Surfactants and additives can be used to raw biomass and biodiesel production technology.
24 K. PIKULA ET AL.

Enzymatic and supercritical extraction and transesterifi- the University of Crete and the University Hospital of Heraklion
cation technologies are capable to replace conventional and also the Chairman of the Division of Morphology of the
Medical School of the University of Crete in Greece. He is teach-
chemical methods and facilitate environmentally
ing the toxicology course for medical students and specialized
friendly, safe, and sustainable biodiesel production. toxicology topics for postgraduate programs in few univer-
The life cycle assessment approach will allow select- sities for 30 years. He also supervised over 30 PhDs. Aristides
ing the most suitable conditions based on a variety of Tsatsakis is Emeritus Professor for the Erisman Russian
feedstock types, methods of biodiesel production, geo- Federal Institute of Hygiene and Toxicology in Moscow
graphical location, social situation, and other character- (2014), Doctor Honoris Causa of the Mendeleev Moscow Uni-
versity in Russia (2016), Doctor Honoris Causa of the Far East
istics. Therefore, the production of biodiesel is an
Federal University, FEFU, in Vladivostok (2017) and Doctor
excellent opportunity for mankind to get a stable, safe, Honoris Causa of the Carol Davila, in Bucharest (2017). He
and renewable source of energy. has written over 1000 publications (books and abstracts pro-
ceedings), 400 of them peer reviewed in prestigious journals.
He is holder of several patents and has an extensive array of
Disclosure statement over 10100 citations and 400000 reads /downloads to his
papers as shown in scientific data bases. Dr Tsatsakis has
No potential conflict of interest was reported by the author(s). given numerous lectures as keynote and has been plenary
speaker in international congresses continuously for the last
20 years.
Funding Dr Kirill Gholokhvast obtained his in 2003 PhD in Physiology
This work was funded by the Russian Science Foundation in the Far Eastern State Agricultural University (Blagovesh-
[grant number: 15-14-20032-P]. chensk, Russia) and in 2017-2019 was Professor of the
Russian Academy of Sciences. In 2018 he was elected as a Cor-
responding Member of the Russian Academy of Education.
Note on contributor Currently he is a Deputy Director for Research, N.I. Vavilov
All-Russian Research Institute of Plant Genetic Resources,
Dr Konstantin Pikula currently works as a senior Researcher in Saint Petersburg, Russian Federation Professor and founder
N.I. Vavilov All-Russian Institute of Plant Genetic Resources: and advisor of SciEdCentre of the Nanotechnology, Far
Saint-Petersburg, RU and Far Eastern Federal University: Vladi- Eastern Federal University (Vladivostok, Russia). He is a
vostok, RU. He obtained his PhD in Far Eastern Federal Univer- member of the editorial board of ‘Food and Chemical Toxi-
sity: Vladivostok, RU. His research interests lie in the area of cology’ (Elsevier), ‘Turkish Journal of Agriculture and Forestry’
ecotoxicology. Currently, his research focuses on the following (Academic Journals), ‘Toxicology Reports’ (Elsevier), ‘Arctic
issues: (1) aquatic nanotoxicology and (2) risk assessment of and North’ (North (Arctic) Federal University), ‘Herald of Kam-
particulate matter emitted during industrial technological chatsk State Technical University’ (Kamchat State Technical
processes. University).
Alexander Zakharenko currently works at the School of Engin-
eering, Far Eastern Federal University. Alexander does research
in Biotechnology, Marine Biology and Immunology. Their ORCID
current project is ’Nanoparticles. Biominerals. Supercritical Konstantin Pikula http://orcid.org/0000-0002-3185-3335
fluids. Polysaccharides’. Alexander Zakharenko http://orcid.org/0000-0002-9520-
Antonios Stratidakis graduated in the Department of 8271
Materials Science, University of Patras, Greece in 2014 and in Mayya Razgonova http://orcid.org/0000-0002-9732-1649
2016 he obtained his Master’s Degree entitled ‘Advanced Poly- Yaroslav Mezhuev http://orcid.org/0000-0003-3075-9010
meric and Nanostructured Materials”, Department of Chem- Aristidis Tsatsakis http://orcid.org/0000-0003-3824-2462
istry, University of Patras, Greece. Until now he has published Kirill Golokhvast http://orcid.org/0000-0002-4873-2281
a number of scientific research articles in International peer
reviewed scientific journals and he has participated in a
number of European and International Conferences with References
several oral and poster presentations. Currently he is a PhD
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