International Journal of Environmental Science and Technology

https://doi.org/10.1007/s13762-019-02279-6

REVIEW

A review of optimum conditions of transesterification process

for biodiesel production from various feedstocks
S. Ishak1 · A. Kamari1

Received: 28 September 2018 / Revised: 10 February 2019 / Accepted: 13 February 2019

© Islamic Azad University (IAU) 2019

Abstract
Development of low-cost, sustainable and environmentally friendly biodiesel is a key focus in the energy industry worldwide.
It is known that selecting suitable raw feedstock materials and optimising an efficient transesterification process are crucial
in biodiesel production. Effective and realistic strategies are imperative in order to boost the quantity and quality of biodiesel
production. In fact, scientists and researchers have put great effort to improve the market value of their biodiesel products. In
recent years, the feasibilities of several raw materials as feedstocks to produce biodiesel have been assessed by researchers.
These materials were reported able to produce biodiesel that met international standards, namely the American Society for
Test and Materials D6751 and European Standard 14214. Although these biodiesel products have met the quality criteria,
their fatty acid methyl ester (FAME) contents differ. This review focuses on optimum conditions (methanol/oil molar ratio,
amount of catalyst, reaction temperature and reaction time) of transesterification process for biodiesel production from vari-
ous feedstocks. The influence of these factors on FAME composition is also discussed. This review is beneficial to scientists
working on biodiesel production particularly for the evolution of eco-friendly and low-cost biodiesel.

Keywords Renewable energy · Biofuel · Biodiesel · Feedstock · Transesterification

Introduction Alternatively, renewable energy is becoming an attractive

Global energy development is influenced by several aspects,
such as fuel availability, energy prices, and economic and
population growth (Gicquell 2013; Khatib 2012; Manzano-
Agugliaro et al. 2011). To date, the world’s energy mix
resources are natural gas, crude oil, hydropower, geothermal,
biomass and coal. Fossil fuel represents the largest energy
source about 78% by value, and the demand for fossil fuel
will increase steadily by 2040 (U.S. Energy Information
Administration 2015). However, the crucial issues for fossil
fuel are the depletion of resources and its ability to produce
harmful greenhouse gases (GHGs) (Bardi 2009; Hook and
Tang 2013).
Editorial responsibility: R Saravanan.
* A. Kamari
azlan.kamari@fsmt.upsi.edu.my; azlkam@yahoo.co.uk
1
Department of Chemistry, Faculty of Science
and Mathematics, Universiti Pendidikan Sultan Idris,
35900 Tanjong Malim, Malaysia
solution in order to decrease the world’s reliance on fossil
fuels as well as to develop cleaner environment (Manzano-
Agugliaro et al. 2012; Sawangkeaw and Ngamprasertsith
2013; Singh and Singh 2010). In fact, the global demand
for energy derived from renewable resources is expected to
continue increase by approximately 2.6% during the period
of 2012 to 2040 (U.S. Energy Information Administration
2015). In recent years, biodiesel becomes one of the most
popular renewable energy resources mainly for transporta-
tion sector. Biodiesel is available in the fuel market in a
number of different blends, including B100 (pure), B20–B30
(blends), B5 (additive) and B2 (lubricity-additive) (Yusuf
et al. 2011). The properties of biodiesel have been well dis-
cussed by Demirbas (2009) and Yusuf et al. (2011). They
also stated that those properties were comparable to that of
petro diesel, which can be applied to diesel engine without
any modification.
Over the years, pyrolysis and transesterification have been
the main processes applied for biodiesel production (Pra-
manik 2003). Heating for pyrolysis is typically conducted
without the presence of air or oxygen, which purposely to
produce smaller molecules through cleavage of chemical

13
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bonds (Canakci et al. 2009; Yusuf et al. 2011). The oil and
fat can be converted into biodiesel by react them with alco-
hol such as MeOH or EtOH (Borges and Díaz 2012). The
conversion process is normally catalysed by NaOH or KOH
(Demirbas 2009). Transesterification offers several advan-
tages such as cost-effectiveness and simplicity in operational
procedure (Baskar and Aiswarya 2016; Salvi and Panwar
2012).
Primarily, transesterification involves reversible reac-
tions to produce intermediate products such as diglycerides
and monoglycerides with the addition of alcohol to shift the
equilibrium (Yusuf et al. 2011; Baskar and Aiswarya 2016;
Sunita et al. 2008). In this context, methanol is the most rec-
ommended alcohol to produce biodiesel (Kirubakaran and
Arul, 2018; Nguyen et al. 2018; Nguyen et al. 2018; Berch-
mans and Hirata 2008). Methanol possesses excellent polar-
ity properties, being the shortest chain alcohol. Studies by
Carvalho et al. (2017) and Mazanov et al. (2016) have shown
that combination of ethanol and heterogeneous catalyst in a
transesterification process had successfully produced 97%
and 97.6% yields of biodiesel, respectively. Verma et al.
(2016a) investigated transesterification process and reported
a higher biodiesel yield of 91.05% for methanol as compared
to 77.4% for ethanol. They also stated that transesterification
using ethanol needs a longer reaction time due to its lower
reactivity and difficulty in separating glycine.
Farobie et al. (2016) reported on the capability of super-
critical 1-propanol in the transesterification process of castor
oil, producing 93.8 mol% yield of biodiesel. However, it was
found that supercritical 1-propanol has lower reactivity than
methanol and ethanol, even though it has a high energy con-
tent. Meanwhile, Verma et al. (2016b) investigated karanja
oil and found the ability of longer-chain alcohols (1-butanol,
1-pentanol and 2-propanol) to give biodiesel in yields of
76.4, 73.13 and 56.36%, respectively. They also reported
that more catalyst and a longer reaction time were required
to accomplish the transesterification reaction. Musa (2014)
has reported on the unsuitability of longer-chain alcohols
Table 1  Comparison Feedstocks
of biodiesel yield using
different types of alcohols for Chicken fat
transesterification process
Jatropha curcas L.
Black soldier fly
Black soldier fly
Mucor circinelloides
Rapeseed oil
Karanja oil
Castor oil
Karanja oil
13
International Journal of Environmental Science and Technology
for transesterification due to their tendency to cause steric
hindrance. These longer-chain alcohols have great ability
to react with water and form azeotropes (Van Gerpen et al.
2004). The comparison of biodiesel yield using different
types of alcohols in the transesterification process is pre-
sented in Table 1.
It is known that there are two types of transesterification
process, namely conventional processes (ultrasound-assisted,
in situ and reactive distillation) and catalytic processes
(homogeneous and heterogeneous) (Fukuda et al. 2001).
Homogeneous catalysts can be categorised into alkali-based
and acid, while heterogeneous catalysts include solid acid
and solid base catalysts (Baskar and Aiswarya 2016; Lin
et al. 2011).
Homogeneous catalysts (acidic) are able to improve the
reaction rate and are preferred due to the simple procedure
and quick completion of the reaction. Generally, the mecha-
nism of homogeneous catalysts will involve nucleophilic
attack on the carbonyl group (Baskar and Aiswarya 2016;
Zhang et al. 2010). Transesterification using homogeneous
catalysts is suitable to be conducted at low temperature but
requires a high concentration of alcohol for high ester yield
(Baskar and Aiswarya 2016; Vicente et al. 2004).
Meanwhile, heterogeneous catalysts (solid catalysts),
either acidic or basic, are favoured for industrial-scale bio-
diesel production. As discussed by Helwani et al. (2009) and
Pinzi et al. (2013), heterogeneous catalysts are tolerant to
high water content and fatty acid, and they are separable and
reusable. In addition, the esterification and transesterifica-
tion reactions can be conducted concurrently with the pres-
ence of an acid catalyst and a small amount of base catalyst.
However, a combination of heterogeneous catalyst and co-
solvent is recommended to enhance the reaction rate (Zabeti
et al. 2009). Furthermore, heterogeneously catalysed reac-
tions generally have a high tendency to form three phases
with alcohol and oil (Sani et al. 2014; Zabeti et al. 2009).
Although there are several reviews on potential biodiesel
feedstocks, they focus mostly on the scenarios and prospects,
Alcohols Biodiesel yield, (%) References
Methanol 90.41 Kirubakaran and Arul (2018)
Methanol 90 Berchmans and Hirata (2008)
Methanol 96.18 Nguyen et al. (2018)
Methanol 94.14 Nguyen et al. (2018)
Ethanol 97 Carvalho et al. (2017)
Ethanol 97.6 Mazanov et al. (2016)
Methanol 91.05 Verma et al. (2016a)
Ethanol 77.4
1-Propanol 93.8 Farobie et al. (2016)
1-Butanol 76.4 Verma et al. (2016b)
1-Pentanol 73.13
2-Propanol 56.86
International Journal of Environmental Science and Technology
characteristics and quality standards, issues and challenges,
food–fuel nexus, engine performance and advance technol-
ogy for biodiesel production. A list of selected review arti-
cles and their specific focus related to biodiesel production
is presented in Table 2. To the best of our knowledge, there
is no review focusing on the optimum conditions of transes-
terification process for different raw materials used as feed-
stocks for biodiesel production. Therefore, in this review,
extensive lists of materials utilised in the production of
biodiesel have been compiled. The optimum conditions for
their transesterification and fatty acid methyl ester (FAME)
content are also discussed.
Feedstocks and optimum transesterification
process for biodiesel production
Biodiesel feedstocks can be classified into: (1) vegetable
oils, (2) waste cooking oils, (3) animal fats, (4) microorgan-
isms and (5) insect larvae. It has been identified that the
feedstock price has greatly influenced the economic viabil-
ity of biodiesel (Atabani et al. 2012; Pinzi et al. 2013). As
discussed by Mofijur et al. (2013) and Pinzi et al. (2013),
the feedstock price could reach 70–95% of the total cost
of biodiesel production. It is therefore crucial to select raw
materials that are cost-effective as feedstocks for biodiesel
production. The ideal raw feedstock must be available in
large quantities and at low cost for large-scale production.
Indeed, the availability of feedstocks depends on the geo-
graphical locations, regional climate, local soil conditions
and agricultural practices of any country (Singh and Singh
2010; Kumar and Sharma 2011).
It has been forecasted that the global production of bio-
diesel was over 26 million tonnes in 2014, of which 40%
was produced by European countries. In 2014, the Argen-
tina, Indonesia, USA, Germany and Brazil were the major
biodiesel producers in the world, with total production of
2.55, 2.75, 4.15, 3.00 and 3.00 million tonnes, respectively
(Souza et al. 2017). Soybean, rapeseed and palm oil were the
main feedstocks used for global biodiesel production in 2013
(Food and Agriculture Organization (FAO) 2015). Soybean
is the main feedstock for the USA, Brazil and Argentina;
meanwhile, in France and Germany biodiesel is produced
mainly from rapeseed. In Asian countries such as Malaysia,
Indonesia and Thailand, palm oil is the main feedstock for
biodiesel production.
Vegetable oils
Vegetable oils are naturally renewable, biodegradable and
involve environmentally friendly procedures (Yusuf et al.
2011; Marchetti et al. 2007). The major component of veg-
etable oil is triacylglycerols (TAGs) or triglycerides (TGs),
which are composed of three fatty acid esters attached to the
glycerol backbone (Issariyakul and Dalai, 2014). Vegetable
oils can be classified into edible and non-edible oils and have
been widely used for biodiesel production as reported by
several studies. Oils studied include coconut oil (Sulaiman
et al. 2013), canola, sunflower and soybean oils (Shumaker
et al. 2007), and jatropha, madhuca, linseed, jojoba, neem,
cottonseed, polanga, tobacco, rapeseed, rubber seed and
karanja oils (Ahmad et al. 2014; Berchmans and Hirata
2008; Puhan et al. 2005; Sahoo and Das 2009; Ghadge and
Raheman 2005). Specifically, the oil seed for biodiesel pro-
duction needs to contain a high volume of oil, and to be easy
to plant on a large scale and able to be harvested the whole
year (Moser and Vaughn 2012). A summary of the studies
using vegetable oils for biodiesel production is presented
in Table 3.
The optimum condition for transesterification process
of coconut waste for biodiesel production was studied by
Sulaiman et al. (2013), and it was found that the biodiesel
yield was to be 88.5% with 2.0 wt.% of potassium hydroxide
(KOH) catalyst at a mixing rate of 700 rpm and reaction
temperature of 62 °C. The biodiesel produced from coconut
waste showed a density of 872.91 kg/m3, an acid value of
0.348 mg/g KOH, viscosity of 3.62 mm2/s, free glycerol of
0.0028 wt.%, total glycerol of 0.0478 wt.%, pour point of
− 5 °C and cloud point of − 2 °C. These properties demon-
strated the suitability of coconut waste combined with in situ
transesterification for biodiesel production. In addition, they
emphasised the application of in situ transesterification to
reduce the cost of materials and reaction time compared to
conventional transesterification processes.
The feasibility and optimum experimental condition for
biodiesel production using kutkura fruit seed were investi-
gated by Kakati and Gogoi (2016). They found that the kut-
kura seed was able to produce up to 70% yield of biodiesel
containing 52.553% of linoleic acid, 30.251% of oleic acid,
7.187% of palmitic acid, 5.382% of stearic acid, 1.449% of
arachidic acid and 1.252% of eicosenoic acid. About 35.45%
of extracted oil containing 3% free fatty acids (FFAs) under-
went a direct base-catalysed transesterification using 3:10 of
methanol/oil with 8 g of sodium hydroxide (NaOH) catalyst
at 60 °C for 2 h. Kutkura seed oil was reported to contain
15.475% of total saturated fatty acids and 84.524% unsatu-
rated fatty acids. Overall, the properties of biodiesel from
kutkura fruit oil met the recommended standards of the
ASTM D6751 and EN 14214, including a calorific value
of 39.717 MJ/kg, a kinematic viscosity of 5.601 mm2/s and
density of 885.3 kg/m3. The aforementioned results con-
firmed that kutkura fruit seed can be used as an alternative
biodiesel feedstock. However, it was noted that this oil was
easily oxidised due to high unsaturated fatty acid content
and hence an oxidation inhibitor was required to increase
biodiesel stability. An engine performance test found that
13
 Table 2  List of selected review articles and their specific focus related to biodiesel production
No. Title of review article Specific focus Reference

1. Recent advancement in biodiesel production methodologies using various Nanocatalytic technology for biodiesel process Ambat et al. (2018)
feedstock: A review
2. A review on ionic liquids as perspective catalysts in transesterification of dif- Development of ionic liquids as effective catalysts for biodiesel preparation Ullah et al. (2018)

13
ferent feedstock oil into biodiesel from various feedstocks
3. Catalysts from renewable resources for biodiesel production Recent development on the heterogeneous catalyst derived from renewable Shan et al. (2018)
resources for the transesterification reaction
4. Biodiesel as alternative fuel for marine diesel engine applications: A review Development of biodiesel as an alternative to marine fuel Noor et al. (2018)
5. A review on the properties, performance and emission aspects of the third The properties of biodiesel fuels from non-edible feedstocks and their perfor- Sakthivel et al. (2018)
generation biodiesel mance on diesel engines
6. Synthesis of biomass as heterogeneous catalyst for application in biodiesel Biodiesel production using biomass as catalyst Tang et al. (2018)
production: State of the art and fundamental review
7. Feedstocks for biodiesel production: Brazilian and global perspectives Current global biodiesel scenarios and selected feedstocks for biodiesel Souza et al. (2017)
production
8. A review on the prospects of sustainable biodiesel production: A global Biodiesel production using waste-oriented oils/fats Hajjari et al. (2017)
scenario with an emphasis on waste-oil biodiesel utilization
9. A review of biomass-derived heterogeneous catalyst for a sustainable bio- Application of bio-based heterogeneous catalyst for biodiesel production Abdullah et al. (2017)
diesel production
10. Trends in catalytic production of biodiesel from various feedstocks Various catalytic technologies and challenges in biodiesel production Baskar and Aiswarya (2016)
11. Investigation of heterogeneous solid acid catalyst performance on low grade Heterogeneous solid acid catalyst for biodiesel production from low grade Mansir et al. (2016)
feedstocks for biodiesel production: A review feedstocks
12. Challenges in biodiesel industry with regards to feedstock, environmental, Issues and challenges in developing a sustainable biodiesel industry in Anuar and Abdullah (2016)
social and sustainability issues: A critical review Malaysia
13. Review of process parameters for biodiesel production from different feed- The various technical aspects of biodiesel production methodology Verma and Sharma (2016)
stocks
14. Prospects of 2nd generation biodiesel as a sustainable fuel-Part: 1 selection of Biodiesel production from selected 2nd generation feedstocks Bhuiya et al. (2016)
feedstocks, oil extraction techniques and conversion technologies
15. A comprehensive review of biodiesel as an alternative fuel for compression The production, performance and emissions from engine using biodiesel Datta and Mandal (2016)
ignition engine
16. Progresses in waste biomass-derived catalyst for production of biodiesel and Biodiesel and bioethanol production using waste biomass-derived catalysts Chakraborty et al. (2016)
bioethanol: a review
17. Effects of biodiesel from different feedstocks on engine performance and The engine performance and emission using biodiesel from different feed- Ghazali et al. (2015)
emissions: A review stocks
18. A review on recent advancement in catalytic materials for biodiesel produc- The varieties of oil feedstock, lipid content, fatty acid composition and bio- Avhad and Marchetti (2015)
tion diesel production process
19. A review of current technology for biodiesel production: State of the art Various technologies for biodiesel production, particularly using heterogene- Aransiola et al. (2014)
ous catalysts
20. Effect of biodiesel from various feedstocks on combustion characteristics, The effects and impacts of biodiesel derived from edible, non-edible and Mofijur et al. (2013)
engine durability and materials compatibility: A review waste cooking oils feedstocks on engine performance
21. Potential of feedstock and catalysts from waste in biodiesel preparation: A The application of waste-based feedstocks and catalysts in biodiesel produc- Nurfitri et al. (2013)
review tion
International Journal of Environmental Science and Technology
 International Journal of Environmental Science and Technology

blending kutkura fruit seed biodiesel with diesel up to a ratio

Borugadda and Goud (2012)

Sawangkeaw and Ngampra-
of 20% was able to reduce volumetric fuel consumption and

Abbaszaadeh et al. (2012)

Shahid and Jamal (2011)

Karmakar et al. (2010)
smoke production.

Atadashi et al. (2012)

Atabani et al. (2012)
sertsith (2013) In another study, Ramadhas et al. (2005) produced
biodiesel using high-FFA rubber seed. The FFA content
of rubber seed oil was first reduced to less than 2% using
Reference

acid-catalysed esterification followed by alkaline-catalysed

transesterification that transformed the products to mono-
esters and glycerol. Moreover, the optimum ester yield was
The synthesis of biodiesel through esterification and transesterification using

Biodiesel production using different methods such as transesterification and

achieved using 9:1 methanol/oil molar ratio and 0.5% NaOH

The physicochemical properties of the plant and animal resources as bio-

at 45 °C for 30 min. The properties of biodiesel from rub-
Potential feedstocks, effective methodologies, properties, quality and the

ber seed were reported as specific gravity of 0.874, calorific

value of 36.50 MJ/kg, kinematic viscosity of 5.81 mm2/s,
flash point of 130 °C, cloud point of 4 °C, pour point of
Methods and technical development in biodiesel production
Biodiesel production from various feedstocks and catalysts

− 8 °C and acid value of 0.118 mg/g KOH. These properties

were found to meet the requirements set by ASTM stand-
ards, and it was highly preferred to blend the biodiesel with
Biodiesel production from lipid-based biomasses

diesel to improve the flash point value for better fuel storage
safety. Therefore, low-cost biodiesel can be produced from
rubber seed, and it was suggested to determine its suitability
in diesel engines over the long term.
Berchmans and Hirata (2008) carried out an optimisation
study on Jatropha curcas L. and observed a reduction in
non-catalytic processes

FFA level to less than 1% using acid-catalysed esterification

non-edible feedstocks

under the optimum experimental conditions of a weight ratio

future of biodiesel

diesel feedstocks

of 0.6:1.0 methanol/oil and 1 wt.% sulphuric acid ­(H2SO4)

Specific focus

for 1 h at 50 °C. Reaction was completed using alkaline-

catalysed transesterification under the optimum conditions:
a molar ratio of 0.24:1.0 methanol/oil and 1.4 wt.% NaOH
at 65 °C for 2 h. Results obtained from gas chromatography/
mass spectrometry (GCMS) analysis showed the fatty acid
28. Properties of various plants and animals feedstocks for biodiesel production
25. A comprehensive review on biodiesel as an alternative energy resource and

profile was: lauric acid (0.06%), myristic acid (0.10%), pal-

23. Biodiesel production from renewable feedstocks: Status and opportunities

mitic acid (14.96%), palmitoleic acid (1.10%), stearic acid

22. A review of lipid-based biomasses as feedstock for biofuels production

(3.85%), oleic acid (32.49%) and linoleic acid (47.43%). Ini-

26. Current biodiesel production technologies: A comparative review

tially, it was reported that J. curcas L. oil contained a high

FFA of about 14.9%; hence, the two-step transesterification
24. Production of biodiesel using high free fatty acid feedstocks

was successfully applied to produce up to 90% yield of bio-

diesel. Furthermore, they suggested the suitability J. curcas
L. as a biodiesel feedstock due to its ability to be easily
grown in tropical and subtropical countries.
27. Production of biodiesel: A technical review

Another study on J. curcas L. by Kamel et al. (2018)

reported its feasibility as a catalyst and oil feedstock for
producing biodiesel. Application of catalyst derived from J.
curcas L., namely: (1) calcined jatropha cake (CJC) and (2)
KOH-activated jatropha cake (KJC), proved the applicabil-
ity of jatropha oil combined with jatropha-derived catalyst
No. Title of review article

in transesterification processes was successful, producing

its characteristics

94.94% and 96.77% yield of biodiesel, respectively. They

Table 2  (continued)

also reported that the optimum conditions for high biodiesel

yield were a methanol/oil molar ratio of 6:1 and 1 wt.%. of
catalyst (CJC and KJC) at 60 °C for 1 h.
Ghadge and Raheman (2006) studied the optimum
experimental conditions for conversion of Madhuca indica

13
 Table 3  Biodiesel production from several vegetable oils as feedstocks
Vegetable oil Process description MeOH/oil Amount of catalyst T, °C t, Biodiesel properties Yield FAME composition, (%) References
molar ratio min

Coconut waste In situ NR 2 wt.%, KOH 62 NR Density: 872.91 kg/m3 88.5% NR Sulaiman et al. (2013)
Acid value: 0.348 mg/g KOH
Viscosity: 3.62 mm2/s

13
Free glycerol: 0.0028 wt.%
Total glycerol: 0.0478 wt.%
Cloud point: − 2 °C
Pour point: − 5 °C
Kutkura fruit seed Alkaline catalysed 3:10 8 wt.%, NaOH 63 120 Calorific value: 39.717 MJ/kg 70% Linoleic acid: 52.553 Kakati and Gogoi (2016)
Kinematic viscosity: 5.601 mm2/s Oleic acid: 30.25
Density: 885.3 kg/m3 Palmitic acid: 7.187
Stearic acid: 5.382
Arachidic acid: 1.449
Eicosenoic acid: 1.252
Rubber seed Acid catalysed 6:1 0.5%, ­H2SO4 50 30 Specific gravity: 0.874 NR NR Ramadhas et al. (2005)
Alkaline catalysed 9:1 5 wt.%, NaOH 45 30 Calorific value: 36.50 MJ/kg
Kinematic viscosity: 5.81 mm2/s
Flash point: 130 °C
Cloud point: 4 °C
Pour point: − 8 °C
Acid value: 0.118 mg/g KOH
Jatropha seed Acid catalysed 0.60% 1%, ­H2SO4 50 60 NR 90% Lauric acid: 0.06 Berchmans and Hirata (2008)
Alkaline catalysed 0.24% 1.4%, NaOH 65 120 Myristic acid: 0.10
Palmitic acid: 14.96
Palmitoleic acid: 1.10
Stearic acid: 3.85
Oleic acid: 32.49
Linoleic acid: 47.43
Jatropha seed Acid catalysed 6:1 1%, ­H2SO4 50 120 NR 94.94% NR Kamel et al. (2018)
Alkaline catalysed 6:1 1%, CJC/KJC 60 60 (CJC)
96.77%
(KJC)
Madhuca indica seed Alkaline catalysed 0.32% 1.24%, ­H2SO4 60 90 Density: 880 kg/m3 98% NR Ghadge and Raheman (2006)
Alkaline catalysed 0.25% 0.7%, KOH 60 60 Viscosity: 3.98 mm2/s
Flash point: 208 °C
Pour point: 6 °C
Water content: 0.04%
Ash content: 0.01%
Carbon residue: 0.20%
Acid value: 0.41 mg/g KOH
Calorific value: 37 MJ/kg
International Journal of Environmental Science and Technology
 Table 3  (continued)
Vegetable oil Process description MeOH/oil Amount of catalyst T, °C t, Biodiesel properties Yield FAME composition, (%) References
molar ratio min

Refined cotton seed Alkaline catalysed 6:1 0.6%, KOH 55 60 Moisture content: 0.020 wt.% 96% NR Onukwuli et al. (2016)
Acid value: 0.22 mg/g KOH
FFA: 0.11%
Saponification value: 165.43 mg/g
Ester value: 165.19 mg/g
Iodine value: 125.28 mg I­ 2/100 g
Peroxide value: 26.01 meq/kg
Specific gravity: 0.8817
Kinematic viscosity: 6.81 mm2/s
Refractive index: 1.344 at 28 °C
Flash point: 173 °C
Cloud point: 7.0 °C
Pour point: 5 °C
Fire point: 193 °C
Cetane number: 56.06
High heating value: 39.54 MJ/kg
Brucea javanica seeds Acid catalysed 6:1 1%, ­H2SO4 65 NR Kinematic viscosity: 3.556 mm2/s 94.34% Palmitic acid: 6.57 Hasni et al. (2017)
Alkaline catalysed NR NaOH NR NR Acid value: 0.027 mg KOH/g Oleic acid: 10
International Journal of Environmental Science and Technology

Cloud point: 2 °C Stearic acid: 49.74

Pour point: 1 °C Linoleic acid: 25.37
Flash point: 164 °C Arachidic acid: 2.21
Cetane index: 51 min Erucic acid: 2.94
Density: 871.3 kg/m3 Others: 2.97
Oxidation stability: 3 h

NR not reported, MeOH methanol, KOH potassium hydroxide, H2SO4 sulphuric acid, NaOH sodium hydroxide, and FAME fatty acid methyl ester

13

seed oil to biodiesel through a two-step transesterification
process. Initially, FFA content was reduced to less than
1% using experimental conditions of methanol/oil volume
ratio of 0.32:1.0 and 1.24 vol.% ­H2SO4 at 60 °C for 1.26 h.
Transesterification was completed using alkaline-catalysed
transesterification with methanol/oil molar ratio of 6:1 and
0.7% w/v KOH. The yield obtained was reported to be
98%. The properties of biodiesel from M. indica seed oil
were: density of 880 kg/m3, acid value of 0.41 mg/g KOH,
viscosity of 3.98 mm2/s, flash point of 208 °C, pour point
of 6 °C, water content of 0.04%, ash content of 0.01%,
carbon residue of 0.20% and calorific value of 37 MJ/kg.
These findings confirmed the suitability of M. indica seed
as a biodiesel feedstock. This study also reported on the
influence on FFA reduction of parameters such as metha-
nol/oil ratio, catalyst concentration and reaction time in
acid-catalysed esterification.
Onukwuli et al. (2016) conducted a study on biodiesel
production using refined cotton seed oil. A biodiesel yield of
up to 96% was reported using alkaline transesterification at
the optimum conditions of a methanol/oil molar ratio of 6:1
and 0.6% KOH at 55 °C for 60 min. The biodiesel properties
produced were reported as: moisture content of 0.020 wt.%,
acid value of 0.22 mg/g KOH, FFA of 0.11%, saponification
value of 165.43 mg/g, ester value of 165.19 mg/g, iodine
value of 125.28 mg ­I2/100 g, peroxide value of 26.01 meq/
kg, specific gravity of 0.8817, kinematic viscosity of
6.81 mm2/s, refractive index of 1.344 at 28 °C, flash point
of 173 °C, cloud point of 7.0 °C, pour point of 5 °C, fire
point of 193 °C, cetane number of 56.06 and a high heating
value of 39.54 MJ/kg. The properties revealed the feasibility
of cotton seed as biodiesel feedstock. It was noted that the
iodine value of the biodiesel showed high unsaturated double
bond content resulting in low oil viscosity.
A study by Hasni et al. (2017) found that Brucea javanica
seed oil was able to produce 94.34% yield of biodiesel in a
two-step transesterification. The acid-catalysed esterifica-
tion was conducted at a methanol/oil ratio of 6:1, catalyst
concentration of 1% and reaction temperature of 65 °C. The
level of FFA was reported to be reduced to 2%. With the
exception of oxidation stability, the properties of biodiesel
from B. javanica seed oil were reported to be comparable to
that of ASTM D6751 and EN 14214 standards. The prop-
erties of biodiesel produced were: acid value of 0.027 mg
KOH/g, kinematic viscosity of 3.556 mm2/s, flash point of
164 °C, cloud point of 2 °C, pour point of 1 °C, cetane index
of 51 min, density of 871.3 kg/m3 and oxidation stability of
3 h. The GCMS profiles of biodiesel from B. javanica seeds
oil were: palmitic acid (6.57%), oleic acid (10%), stearic acid
(49.74%), linoleic acid (25.37%), arachidic acid (2.21%),
erucic acid (2.94%) and others (2.97%). These results
showed the capability of B. javanica seeds to be a biodiesel
feedstock. Nevertheless, in order to overcome the issue of
13
International Journal of Environmental Science and Technology
low biodiesel stability, an antioxidant (propyl gallate) had to
be used to raise the oxidation stability to 70 h.
Waste cooking oils
Basically, waste cooking oils are produced in huge quantities
daily as a result of the tremendous growth in human popula-
tion. It can easily be collected from restaurants and houses.
Direct discharge of waste cooking oil into the public drain-
age system may create negative impacts on the environment,
such as long-term soil pollution, toxicity to aquatic life and
effects on human health; it was reported that the use of recy-
cled waste cooking oils may cause severe health problems
such as cancer (Yaakob et al. 2013). Recycled waste cooking
oils will normally undergo oxidation, which later produces
carcinogenic compounds that may harm human health. Thus,
it is preferable to apply waste cooking oils to biodiesel pro-
duction, as it offers a low raw feedstock cost and avoids
controversy in food supply as compared to vegetable oils
(Yaakob et al. 2013; Phan and Phan 2008). A description of
studies using waste cooking oils for biodiesel production is
summarised in Table 4.
The capability and optimum condition for heterogene-
ous transesterification were investigated by Omar and
Amin (2011) using waste palm cooking oil in the presence
of a heterogeneous catalyst, namely strontium zirconia (Sr/
ZrO2). It was reported that operating parameters had signifi-
cant effects on the transesterification. The highest biodiesel
yield (79.7%) was produced using optimum conditions of
methanol/oil molar ratio of 29:1 and 2.7 wt.% of Sr/ZrO2
for 87 min at 115.5 °C. Therefore, this study supports the
application of waste cooking oil as a biodiesel feedstock
with the assistance of Sr/ZrO2. An optimisation study was
conducted and found that the most significant variables for
high FFA conversion and biodiesel yield were the reaction
time and the temperature.
Cai et al. (2015) studied the effect of chemical treatment
on waste cooking oil for biodiesel production. The FFA
conversion was reported to be up to 99.6% with acid value
of 124.9 mg KOH/g. Acid-catalysed esterification was con-
ducted using a glycerol/FFA molar ratio of 1.4:1 for 4 h at
210 °C and catalyst loading (soap) of 0.5 wt.%. The esterified
waste cooking oil was then subjected to alkaline-catalysed
transesterification using methanol/oil molar ratio of 6:1 cata-
lysed by 0.33% NaOH for 1 h at 40 °C to give the optimum
biodiesel content of 98.6 wt.%. The properties of biodiesel
from this transesterification were reported to be 98.6% of
ester content, kinematic viscosity of 4.603 mm2/s at 40 °C,
density of 874.9 kg/m3, cetane number of 64.2, flash point
of 134 °C, 0.001 wt.% free glycerine, 0.019 wt.% of total
glycerine, acid value of 0.6 mg KOH/g, oxidation stability
of 15.9 h at 110 °C and sulphur content of 12.7 mg/kg. The
FAME profiles were reported as: myristic acid (1.1 ± 0.16%),
 Table 4  Biodiesel production from several waste cooking oils as feedstocks
Process MeOH/oil molar Amount of catalyst T, °C t, min Biodiesel properties Yield FAME composition, % References
description ratio

Heterogeneous transesteri- 29:1 2.7 wt%, Sr/ZrO2 115.5 87 NR 79.7% biodiesel NR Omar and Amin
fication (2011)
Acid catalyst esterification 1.4:1 0.5%, soap 210 240 Ester content: 98.6% 93.1% biodiesel Myristic acid: Cai et al. (2015)
Alkaline transesterification 6:1 0.33%, NaOH 40 60 Kinematic viscosity: 4.603 mm2/s 1.1 ± 0.16
Density: 874.9 kg/m3 Palmitic acid: 37.5 ± 1.4
Cetane number: 64.2 Palmitoleic acid: 1.7 ± 0.10
Flash point: 134 °C Stearic acid: 6.2 ± 0.50
Free glycerine: 0.001 wt.% Oleic acid:
Total glycerine: 0.019 wt.% 38.7 ± 1.81
Acid value: 0.6 mg KOH/g Linoleic acid: 13.1 ± 0.55
Oxidative stability: 15.9 h, 110 °C Linolenic acid: 0.75 ± 0.14
Sulphur content: 12.7 mg/kg Arachidic acid: 0.55 ± 0.04
International Journal of Environmental Science and Technology

Eicosenoic acid: 0.40 ± 0.09

Heterogeneous transesteri-
fication
Heterogeneous transesteri-
fication
Acid catalyst esterification
Alkaline catalyst transesteri- 1:3
fication
70:1 10 wt.%, heteropoly 65 840 Acid value: 0.493 mg KOH/g 88.6% FFA conversion NR Talebian-Kiakalaieh
acid Kinematic viscosity: 5.24 mm2/s et al. (2013)
Density: 867.2 kg/m3
Flash point: 168 °C
Cloud point: − 1 °C
Pour point: − 4.4 °C
10:1 8 wt.%, ­FeCl3 90 120 Density at 20 °C: 0.87 g/mL 92% biodiesel Palmitic acid: 27.35 Ma et al. (2017)
Viscosity at 40 °C: 4.4 mm2/s Linoleic acid: 30.95
Acid value: 0.06 mg KOH/g Oleic acid: 24.46
Heating value: 40.47 MJ/kg Stearic acid: 2.55
Cetane number: 50 Hexadecenoic acid: 1.35
Free glycerol content: 0.009 wt.%
Total glycerine content: 0.067 wt.%
Carbon residue: 0.09 wt.%
1:2.5 1%, ­H2SO4 60 320 Specific gravity: 0.8743 94% biodiesel NR Sahar et al. (2018)
1%, KOH 60 50 Viscosity: 5.83 mm2/s
Calorific value: 37.2 kJ/g
Cloud point: 10.5 °C
Pour point: 1 °C
Acid value: 0.6 mg KOH/g
Saponification value: 280 mg KOH/g
Iodine value: 63.5 g ­I2/100 g
Cetane number: 51.48

NR not reported, MeOH methanol, KOH potassium hydroxide, NaOH sodium hydroxide, FAME fatty acid methyl ester, Sr/ZrO2 strontium zirconia

13

palmitic acid (37.5 ± 1.4%), palmitoleic acid (1.7 ± 0.10%),
stearic acid (6.2 ± 0.50%), oleic acid (38.7  ± 1.81%), linoleic
acid (13.1 ± 0.55%), linolenic (0.75 ± 0.14%), arachidic acid
(0.55 ± 0.04%) and eicosenoic acid (0.40 ± 0.09%). These
findings confirmed that waste cooking oil is a favourable
biodiesel feedstock. They reported that soap could be a
potential catalyst and play an important role in high FFA
reduction, leading to high biodiesel yield.
The heterogeneous transesterification of waste cooking
oil using a heteropoly acid was investigated by Talebian-
Kiakalaieh et al. (2013). Based on an optimisation study,
they reported that up to 88.6% conversion of FFA could
be achieved using methanol/oil molar ratio of 70:1 with
addition of 10% catalyst for 14 h at 65 °C. The biodiesel
properties were: acid value of 0.493 mg KOH/g, kinematic
viscosity of 5.24 mm2/s, density of 867.2 kg/m3, flash point
of 168 °C, cloud point of − 1 °C and pour point of − 4.4 °C.
These results proved the capability of waste cooking oil with
the assistance of heteropoly acid for producing biodiesel.
Meanwhile, Ma et al. (2017) investigated biodiesel pro-
duction from waste cooking oil supported by a heterogene-
ous catalyst, namely ­FeCl3-modified resin. They reported a
yield of 92% biodiesel successfully produced under optimum
conditions of a methanol/oil molar ratio of 10:1, catalyst
content 8% and reaction temperature 90 °C for 120 min.
The properties of the biodiesel produced were character-
ised and reported as: density of 0.87 g/mL at 20 °C, viscos-
ity of 4.4 mm2/s at 40 °C, acid value of 0.06 mg KOH/g,
heating value of 40.47 MJ/kg, cetane number of 50, free
glycerol content of 0.009 wt.%, total glycerine content of
0.067 wt.% and carbon residue of 0.09 wt.%. The FAME
profiles were: palmitic acid (27.35%), linoleic acid (30.95%),
oleic acid (24.46%), stearic acid (2.55%) and hexadecenoic
acid (1.35%). Based on biodiesel properties, it was found
that waste cooking oil, with the use of ­FeCl3-modified resin,
could be an alternative cheaper raw material for biodiesel
production. Moreover, it was noticed that ­FeCl3-modified
resin offered several advantages in terms of reusability and
operational stability during the transesterification process.
A study by Sahar et al. (2018) was carried out to deter-
mine the applicability and optimal conditions for trans-
esterification process of waste cooking oil for producing
biodiesel. They reported on the use of two-step transes-
terification due to the high acid value of waste cooking oil
(5.5 mg KOH/g). In this study, the transesterification pro-
cess was initiated with acid-catalysed esterification: meth-
anol oil molar ratio 1:2.5, ­H2SO4 catalyst (1%) at 60 °C for
up to 320 min followed by alkaline-catalysed transesterifi-
cation: methanol/oil molar ratio 1:3, KOH catalyst (1%) at
60 °C for 50 min, to produce a biodiesel yield of 94%. The
biodiesel properties were determined and reported: spe-
cific gravity of 0.8743, viscosity of 5.83 mm2/s, calorific
value of 37.2 kJ/g, cloud point of 10.5 °C, pour point of
13
International Journal of Environmental Science and Technology
1 °C, acid value of 0.6 mg KOH/g, saponification value of
280 mg KOH/g, iodine value of 63.5 g I­ 2/100 g and cetane
number of 51.48.
Animal fats
Animal fats such as chicken fat, lard and tallow are consid-
ered to have high potential for use as feedstocks for produc-
tion of biodiesel (Jeong et al. 2009; Alptekin and Canakci
2011; Araújo et al. 2010). These materials are normally dis-
posed of in landfills in huge quantities (Chakraborty et al.
2014; Marulanda et al. 2010; Sawangkeaw and Ngampra-
sertsith 2013). A summary of studies of the use of animal
fats as feedstocks for biodiesel production is described in
Table 5.
A study using lard for biodiesel production was carried
out by Jeong et al. (2009). They reported that the lard was
successfully converted in a yield of 98.6%. They conducted
the conversion process at optimum conditions of an oil/
methanol molar ratio of 7.5:1, catalyst amount of 1.26%,
reaction temperature of 65 °C and reaction time of 20 min.
The FAME composition in biodiesel produced from lard was
reported as oleic acid (44.7%), palmitic acid (26.4%), stearic
acid (12.1%), linoleic acid (12.7%), linolenic acid (1.0%) and
others (2.1%). They reported several properties of biodiesel
from lard: acid value of 0.08 mg KOH/g, viscosity of 4.2
cP at 40 °C, iodine number of 72 g ­I2/100 g and cold filter
plugging point of 8.0 °C. In spite of its ability to produce
biodiesel, the cold filter plugging point of lard exceeded the
permissible limit. To overcome this issue, they suggested to
blend the biodiesel produced with petrodiesel and additives.
Optimisation of transesterification using chicken fat as a
raw feedstock was studied by Alptekin and Canakci (2011).
The maximum yield was reported as 88.5% for a reaction
performed for 1 h at 60 °C with potassium methoxide
(KOMe) as a catalyst. The fuel produced had properties of
density of 883–891 kg/m3, viscosity of 4.94–6.84 mm2/s,
flash point of 169.9–172.8 °C, methanol content of
0.01–0.20%, pour point of 2–4 °C, heat of combustion of
40,000 kJ/kg and acid value of 0.30 mg KOH/g. It was
reported that the pour point of the biodiesel derived from
chicken fat was found to be slightly higher than the stand-
ard limit, suggesting that the biodiesel has poor cold-flow
behaviour properties. In addition, they also recommended to
conduct a study on the effect on biodiesel yield of potassium
methoxide and sodium methoxide as catalysts.
A study on beef tallow for biodiesel production by
Araújo et al. (2010) obtained 96.26% yield of biodiesel.
Transesterification of 100 g of beef tallow was car-
ried out by adding about 10 mL of methanol followed
by 0.2 g of KOH at temperature range of 70–90 °C for
45 min. The FAME composition of beef tallow bio-
diesel was reported as: lauric acid (0.060 ± 0.00%),
 Table 5  Biodiesel production from several animal fats as feedstocks
Animal fat MeOH/oil Amount of T, °C t, min Biodiesel properties Yield FAME composition, (%) References
molar ratio catalyst

Lard 7.5:1 1.26% 65 20 Acid value: 0.08 mg KOH/g 98.6% Oleic acid: 44.7 Jeong et al. (2009)
Viscosity: 4.2 cP at 40 °C Palmitic acid: 26.4
Iodine number: 72 g ­I2/100 g Stearic acid: 12.1
Cold filter plugging point: 8.0 °C Linoleic acid: 12.7
Linolenic acid: 1.0
Others: 2.1
Chicken fat Acid 40:1 100%, ­H2SO4 60 80 Density: 883 to 891 kg/m3 88.5% NR Alptekin and
catalysed Viscosity: 4.94 to 6.84 mm2/s Canakci (2011)
Alkaline catalysed 6:1 1%, KOMe 60 240 Flash point: 169.9 to 172.8 °C
Methanol content: 0.01 to 0.20%
Pour point: 2 to 4 °C
Combustion heat: 40,000 kJ/kg
Acid value: 0.30 mg KOH/g
International Journal of Environmental Science and Technology

Beef tallow 1:10 15 mL/0.2 g, KOH 70 to 90 45 Acidity index: 0.495 ± 0.007 mg KOH/g 96.26% Lauric acid: 0.06 ± 0.00 Araújo et al. (2010)
Specific weight: 880 ± 0.000 at 20 °C kg/m3 Myristic acid: 2.69 ± 0.28
Flash point: more than 210 °C 12-methyl-tetradecanoic acid: 1.14 ± 0.20
Kinematic viscosity: 5.470 ± 0.005 at 40 °C 14-methyl-pentadecanoic acid: 1.65 ± 0.01
Oxidative stability: 1.990 ± 0.000 h 6,7-methylene-octadecanoic acid: 1.06 ± 0.10
Ester index: 96.67 ± 0.010% mass Palmitic acid: 22.30 ± 0.33
Stearic acid: 41.28 ± 0.23
Oleic acid: 28.82 ± 0.22
Linoleic acid: 0.58 ± 0.20
Arachidic acid: 0.42 ± 0.02
Rohu, Labeo rohita 0.5:1 0.75%, KOH 55 60 Acid value: 0.25 ± 0.04 mg KOH/g 95.9% Palmitic acid: 25.2 Kudre et al. (2017)
Iodine number: 117.0 ± 2.0 g ­I2/100 g Linolenic acid: 11.7
Viscosity: 3.61 ± 1.9 cP at 25 °C Palmitoleic acid: 9.6
Myristic acid: 7.2
Oleic acid: 5.9
Linoleic acid: 6.8
Stearic acid: 5.5
Chicken fat 1:13 8.5 wt.%, chicken 57.5 300 Acid value: 0.4 mg KOH/g 90.14 NR Kirubakaran and
eggshell Density at 15 °C: 926 kg/m3 Arul (2018)
Kinematic viscosity: 4.9 mm2/s at 40 °C
Flash point: 177 °C
Fire point: 185 °C
Lower heating value: 39,175 kJ/kg
Cetane index: 61
Iodine number: 50.24 g ­I2/100 g
Water content: 0.04 vol. %
Molar mass: 952.3 g/mol

NR not reported, MeOH methanol, KOMe potassium methoxide, KOH potassium hydroxide, H2SO4 sulphuric acid, FAME fatty acid methyl ester

13

myristic acid (2.69 ± 0.28%), 12-methyl-tetradeca-
noic acid (1.14 ± 0.20%), 14-methyl-pentadecanoic
acid (1.65 ± 0.01%), 6,7-methylene-octadecanoic
(1.06 ± 0.10%), palmitic acid (22.30 ± 0.33%), stearic
acid (41.28 ± 0.23%), oleic acid (28.82 ± 0.22%), linoleic
acid (0.58 ± 0.20%) and arachidic acid (0.42 ± 0.02%). The
properties of biodiesel from beef tallow were reported:
acidity index of 0.495 ± 0.007 mg KOH/g, specific weight
of 880.0 ± 0.000 kg/m3 at 20 °C, flash point of higher than
210 °C, kinematic viscosity of 5.470 ± 0.005 at 40 °C,
oxidative stability of 1.990 ± 0.000 h and ester index of
96.67 ± 0.010% mass. Despite showing acceptable engine
fuel criteria, stability of the biodiesel was difficult to
achieve because of low oxidation stability. Therefore, an
oxidant test was conducted, which found that the addi-
tion of an antioxidant, butylated hydroxytoluene (BHT),
at 400 ppm, was able to increase the oxidation stability
to 6.18 h.
Kudre et al. (2017) studied the potential of Rohu (Labeo
rohita) fish oil for biodiesel production. The biodiesel
yield was reported as 95.2% at optimum conditions of
0.75% KOH, methanol/oil volume ratio of 0.5:1 at 55 °C
and reaction time of 60 min. They reported the physico-
chemical properties of biodiesel produced: acid value
0.25 ± 0.04 mg KOH/g, iodine number 117.0 ± 2.0 g
­I2/100 g and viscosity 3.61 ± 1.9 cP at 25 °C. Meanwhile,
the FAME profile comprised palmitic acid (25.2%), lino-
lenic acid (11.7%), palmitoleic acid (9.6%), myristic acid
(7.2%), oleic acid (5.9%), linoleic acid (6.8%) and stearic
acid (5.5%). They conducted characterisation studies and
confirmed the successful transformation of Rohu fish oil
to biodiesel based on Fourier transform infrared (FTIR)
and thin-layer chromatography (TLC) analyses. Moreover,
thermal stability tests were also conducted using differen-
tial scanning calorimetry (DSC) and acceptable cold-flow
properties were found. An antioxidant test was also carried
out and revealed the efficiency of butylated hydroxytolu-
ene (BHT) at 300 mg/kg in increasing biodiesel stability
to 30 days.
Meanwhile, Kirubakaran and Arul (2018) conducted a
study on production of biodiesel from chicken fat incorpo-
rating chicken eggshell (catalyst). Initially, esterification of
chicken fat oil was carried out to reduce the acid number
below 1 mg KOH/g of oil, followed by alkaline-catalysed
transesterification. Optimisation studies using response
surface methodology found that the optimum conditions
were: a methanol/oil molar ratio of 1:13 and 8.5 wt.% cata-
lyst at 57.5 °C for 5 h, giving a 90.14% yield of biodiesel.
They reported the biodiesel properties as: acid value 0.4 mg
KOH/g of oil, density of 926 kg/m3 at 15 °C, kinematic vis-
cosity of 4.9 mm2/s at 40 °C, flash point of 177 °C, fire point
of 185 °C, lower heating value of 39,175 kJ/kg, cetane index
of 61, iodine number of 50.24 g I­ 2/100 g, water content of
13
International Journal of Environmental Science and Technology
0.04 vol.% and molar mass 952.3 g/mol. Moreover, engine
tests were carried out for this biodiesel, which confirmed its
suitability, with lower carbon monoxide and hydrocarbon
emissions and filter smoke number compared to diesel.
Microorganisms
Microorganisms such as yeasts, microalgae, bacilli and fungi
are also available for the production of biodiesel due to their
ability to accumulate the large quantities of cellular lipids
(Canakci and Sanli 2008; Chisti 2007; Leong et al. 2015;
Sánchez et al. 2013). A brief explanation of the studies into
using microorganisms for biodiesel production is summa-
rised in Table 6.
The characteristics of microalgae as unicellular photosyn-
thetic organisms make them useful for biodiesel production.
Microalgae produce biomass that contains approximately
50% carbon by using carbon dioxide and energy from light.
The growth rate of microalgae that contributes to lipid con-
tent is highly dependent on organic carbon, light intensity,
pH, agitation speed, nitrogen sources, temperature, salinity
(for marine strains) and dissolved oxygen level. Approxi-
mately 180 tonnes of carbon dioxide is produced by about
100 tonnes of microalgae biomass under either natural or
artificial light. The microalgae also potentially utilise car-
bon dioxide from industrial plants for biodiesel production
(Converti et al. 2009; Miao and Wu 2006; Sawangkeaw and
Ngamprasertsith 2013).
Meanwhile, most yeast and fungi are able to accumu-
late and generate lipids in their cells in the absence of light,
depending on the carbon source from the substrate. It has
been reported that lipids from yeasts and fungi have similar
fatty acid properties to those of soybean and sunflower oils
(Sawangkeaw and Ngamprasertsith 2013; Xia et al. 2011).
Bacteria also have great potential as biodiesel feedstock due
to their characteristics of high lipid content and fast growth
in the dry cell. These characteristics mean that the produc-
tivity of bacteria can be considered close to those of ole-
aginous microorganisms (Adewale et al. 2015; Sawangkeaw
and Ngamprasertsith 2013).
Angerbauer et al. (2008) studied the lipid accumulation of
Lipomyces starkeyi after ultrasonic treatment. They obtained
a maximum lipid content of 68% under conditions of a car-
bon/nitrogen (C/N) ratio of 150:1 in a pH range of 5.0 to
7.5. It was also reported that the reduction in C/N ratio from
150:1 to 60:1 produced only 40% lipid content, highlight-
ing the significant effect of C/N ratio on lipid accumula-
tion. Initially, they digested raw sewage sludge with acid and
alkaline procedures, followed by ultrasonic treatment with
40% maximum power over 3 h. These results revealed the
potential of lipid accumulation by L. starkeyi from sewage
sludge as a biodiesel feedstock. It was suggested that ultra-
sonic was the most suitable pre-treatment of sewage sludge
 Table 6  Biodiesel production from several microorganisms as feedstocks
Microorganisms MeOH/oil Amount of catalyst T, °C t, min Biodiesel properties Yield FAME composition, (%) References
molar ratio

Lipomyces starkeyi NR
Chlorella protothecoides 56:1 100 wt.%, ­H2SO4
Free fatty acids: 12.18% 70% (Lipid) Myristic acid: 0.90 Angerbauer et al. (2008)
Phosphorus content: 7 mg/kg Linolenic acid: 0.12
Sulphur content: 12 mg/kg Palmitic acid: 55.93
Arachidic acid: 0.48
Palmitoleic acid: 1.85
Gadoleic acid: 0.18
Stearic acid: 13.8
Behenic acid: 0.48
Oleic acid: 25.89
Linoleic acid: 0.1
30 240 Density: 0.864 kg/L 55% (Lipid) NR Miao and Wu (2006)
Viscosity: 5.2 mm2/s
Flash point: 115 °C
Cold filter plugging point: − 11 °C
Solidifying point: − 12 °C
Heating value: 41 MJ/kg
Acid value: 0.374 mg KOH/g
International Journal of Environmental Science and Technology

H/C ratio: 1.81

Isochrysis galbana 12:1 1 wt.%, NaOH 62 180 Iodine index: 47.1 g ­I2/100 g 12.5% (FAME) Myristic acid: 18.8 ± 0.16 Sánchezet al. (2013)
Molecular weight: 288.05 g/mol Palmitic acid: 16.5 ± 1.45
Oleic acid: 18.5 ± 1.66
Linoleic acid: 15.6 ± 1.13
Rhodosporidium kratoch- NR 53.18% (Lipid) Palmitic acid: 43.06 ± 0.12 Patel et al. (2015)
vilovae Stearic acid: 28.74 ± 0.32
Oleic acid: 17.34 ± 0.27
Chlorella sp. MJ 11/11 5:1 4 M, HCI 65 420 Kinematic viscosity: 4.6 mm2/s 95% (Lipid) Palmitic acid: 21.5 ± 1.1 Ghosh et al. (2017)
Density: 886 kg/m3 Stearic acid: 4.7 ± 0.2
Calorific value: 39.3 MJ/kg Oleic acid: 32.6 ± 1.2
Iodine value: 80.6 g ­I2/100 g Linoleic acid: 19.2 ± 0.5
Acid value: 0.7 mg/KOH g Linolenic acid: 9.4 ± 0.6
Saponification value: 244.8 mg/KOH g Others: 2.1–2.9 ± 0.6
Flash point: 113 °C
Pour point: − 12 °C
Cloud point: − 2.2 °C
Cetane number: 56.1
Ash content: 0.01% w/w
Water content: 0.04% w/w

13
 Table 6  (continued)
Microorganisms MeOH/oil Amount of catalyst T, °C t, min Biodiesel properties
molar ratio
Mucor circinelloides 120:1 10 wt.%, ­H3PMo12O40 200 240 Saturated FA: 33.62%
Mono-unsaturated FA: 28.98%
Polyunsaturated FA: 33.13%
13
Degree of unsaturation: 95.24
Saponification value: 201.65 mg/g
Iodine value: 104.02 g ­I2/100 g
Cetane number: 49.96
Long-chain saturated factor: 4.8
Cold filter plugging point: − 1.3 °C
Cloud point: 4.96 °C
Pour point: − 1.3 °C
Allylic position equi.: 94.74
Bis-allylic position equi.: 53.17
Oxidation stability: 6.1 h
Higher heating value: 37.51 MJ/kg
Viscosity: 3.2 mm2/s
Density: 850 kg/m3
Cyanobium sp. Limnothrix NR Lipid-Saturated FA: 47.88–86.22%
sp. Nostoc sp. Mono-unsaturated FA: 13.00–46.55%
Poly unsaturated FA: 0.68–3.65%
Cetane number: 51–68.94
Oxidation stability: 6 min–176.01 h
NR not reported, KOH potassium hydroxide, H2SO4 sulphuric acid, NaOH sodium hydroxide, HCI hydrochloric acid, H
­ 3PMo12O40 12-molybdophosphoric acid, FAME fatty methyl ester
Yield FAME composition, (%) References
97% (FAME) Oleic acid: 32.4 Carvalho et al. (2017)
Linoleic acid: 16.34
Linolenic acid: 22.7
Palmitic acid: 15.6
Stearic acid: 3.9
Myristic acid: 1.5
Lauric acid: 0.4
Capric acid: 0.2
Caprylic acid: 0.07
0.43 to 9.12% Lipid- Caprylic acid: 0.06–0.60 Oliveira et al. (2018)
(Lipid) Capric acid: 0.14 –0.83
Lauric acid: 9.99–41.91
Myristic acid: 8.87 –29.48
Pentadecanoic acid: 0.10–0.29
Palmitic acid: 12.90–25.24
Margaric acid: 0.00–0.14
Stearic acid: 3.32–5.86
Arachidic acid: 0.11–0.98
Palmitoleic acid: 0.21–5.94
Oleic acid: 11.51–36.23
Linoleic acid: 0.68–3.65
International Journal of Environmental Science and Technology
International Journal of Environmental Science and Technology
compared to other treatments such as thermal, alkaline and
acid processes. The saturated fatty acid content in this lipid
was determined as 70%, with palmitic acid (55.93%), oleic
acid (25.89%) and stearic acid (13.8%) dominant, giving a
high cetane number which is efficient for engine combustion,
although it showed poor low-temperature behaviour.
The potential and optimum experimental condition
for transesterification process of Chlorella protothe-
coides for biodiesel production were assessed by Miao
and Wu (2006). The percentage of lipid extracted from
these microalgae was reported as 55%. The extracted lipid
underwent an acid-catalysed transesterification under
conditions of a methanol/oil molar ratio of 56:1 at 30 °C
for 4 h. They also reported that the specific gravity of
the biodiesel produced was reduced from 0.912 to a final
value of 0.8637. Biodiesel from C. protothecoides showed
standard properties of density of 0.864 kg/L, viscosity of
5.2 mm2/s, flash point of 115 °C, cold filter plugging point
of − 11 °C, solidifying point of − 12 °C, heating value of
41 MJ/kg, acid value of 0.374 mg KOH/g and H/C ratio of
1.81. Based on these biodiesel properties, they conducted
a comparative study between biodiesel derived from C.
protothecoides and diesel.
Meanwhile, Sánchez et al. (2013) studied Isochrysis
galbana (T-ISO) and found that the optimum harvest bio-
mass concentration of 0.305 g/L with a FAME content of
12.5 was achieved using a methanol/oil molar ratio of 12:1
and 1% NaOH for 3 h at a reaction temperature of 62 °C.
They conducted GCMS analysis, and the major fatty acid
profiles consisted of myristic acid (18.8 ± 0.16%), palmitic
acid (16.5 ± 1.45%), oleic acid (18.5 ± 1.66%) and linoleic
acid (15.6 ± 1.13%). The biodiesel produced was reported
to have an iodine index of 47.1 g I­ 2/100 g of sample and a
molecular weight of 288.05 g/mol. These findings confirm
the suitability of T-ISO for biodiesel production. However,
they reported that the biodiesel produced was susceptible to
oxidation due to the presence of linolenic and other polyun-
saturated fatty acids. To avoid this problem, an additional
pre-treatment known as partial catalytic hydrogenation was
suggested.
The feasibility of Cassia fistula L. fruit pulp and ole-
aginous yeast Rhodosporidium kratochvilovae HIMPA1 to
produce biodiesel was investigated by Patel et al. (2015).
They found lipid accumulations with large intracellular
lipid droplets of 4.35 ± 0.54 µm. Fruit pulp aqueous extract
was used to give total lipids of 4.86 ± 0.54 g/L with a lipid
content of 53.18% (w/w). The FAME profiles showed the
major fatty acids were stearic acid (28.74 ± 0.32%), oleic
acid (17.34 ± 0.27%) and palmitic acid (43.06 ± 0.12%).
These results support the potential of R. kratochvilovae
HIMPA1 grown in Cassia fistula L. fruit pulp as a feedstock
for biodiesel production. However, the biodiesel produced
was prone to auto-oxidation due to the presence of the poly-
unsaturated fatty acid linoleic acid of 0.48 ± 0.011%.
Ghosh et al. (2017) conducted an optimisation study on
the single-step transesterification, using hydrochloric acid
as a catalyst, of algal biomass (Chlorella sp. MJ 11/11) for
biodiesel production. It was found that the lipid conversion
was about 95% at a catalyst concentration of 4 M, an algal
biomass/methanol ratio of 1:5, reaction temperature 65 °C,
reaction time 420 min and biomass drying time of 90 min,
The FAME profiles comprised palmitic acid (21.5 ± 1.1%),
stearic acid (4.7 ± 0.2%), oleic acid (32.6 ± 1.2%), linoleic
acid (19.2 ± 0.5%), linolenic acid (9.4 ± 0.6%) and oth-
ers (2.1–2.9 ± 0.6%). The biodiesel properties were: kin-
ematic viscosity of 4.6 mm2/s at 40 °C, density of 886 kg/
m3 calorific value of 39.3 MJ/kg, iodine value of 80.6 g
­I2/100 g, acid value of 0.7 mg KOH/g, saponification value
of 244.8 mg KOH/g, flash point of 113 °C, pour point of
− 12 °C cloud point of − 2.2 °C, cetane number of 56.1,
ash content of 0.01% w/w and water content of 0.04% w/w.
Based on these findings, the aforementioned algal biomass
was proven able to produce biodiesel, with properties com-
parable to diesel. However, again the issue of biodiesel sta-
bility arose due to presence of unsaturated fatty acids, which
were reported as 68.8 ± 0.2%.
Carvalho et al. (2017) produced biodiesel using Mucor
circinelloides as a raw feedstock and catalysed the conver-
sion reaction by using the solid heteropoly acid 12-molyb-
dophosphoric acid ­H3PMo12O40. They reported that about
40.5% of lipid was extracted during fermentation of M. cir-
cinelloides after up to 120 h of reaction. Moreover, about
97% of biodiesel was produced under experimental condi-
tions of an ethanol/oil molar ratio of 120:1, H
­ 3PMo12O4 con-
centration of 10 wt% and reaction temperature 200 °C, and
continuously stirred at 300 rpm for 4 h. The properties of the
biodiesel produced from M. circinelloides operated in biore-
actor were: saturated fatty acid (33.62%), mono-unsaturated
fatty acids (28.98%), polyunsaturated fatty acids (33.13%),
saponification value (201.65 mg/g), iodine value (104.02 g
­I2/100 g), cetane number (49.96), long-chain saturated fac-
tor (4.8), cold filter plugging point (–1.3 °C), cloud point
(4.96 °C), pour point (–1.3 °C), allylic position equivalent
(94.74), bis-allylic position equivalent (53.17), oxidation sta-
bility (6.1 h), higher heating value (37.51 MJ/kg), viscosity
(3.2 mm2/s) and density (850 kg/m3). Based on GCMS anal-
ysis, the dominant fatty acid ethyl esters (FAEEs) contained
in the biodiesel were reported as oleic acid (32.4%), linoleic
acid (16.34%), linolenic acid (22.7%), palmitic acid (15.6%),
stearic acid (3.9%), myristic acid (1.5%), lauric acid (0.4%),
capric acid (0.2%) and caprylic acid (0.07%). Results from
this study revealed the feasibility of M. circinelloides treated
with 12-molybdophosphoric acid impregnated with alumina
as a feedstock for biodiesel production. They concluded that
direct transesterification was the most preferred technique
13

compared to two-step transesterification, due mainly to the
shorter reaction time which excludes the cell disruption
and oil extraction process. Additionally, they performed
1
H NMR analysis in order to characterise the appearance
of FAEEs.
A study by Oliveira et al. (2018) investigated the bio-
diesel production from three types of cyanobacterial strain:
Cyanobium sp. CACIAM06, Limnothrix sp. CACIAM10 and
Nostoc sp. CACIAM19. These species were cultured in two
different culture media, namely ASM-1 and BG-11. Of these
strains, Limnothrix sp. cultured in ASM-1 showed the high-
est lipid yield (9.12%). They found that 15 types of fatty
acids were identified in the lipids extracted, which contain
high saturated fatty acids in the range from 47.88 to 86.22%,
of which the dominant were lauric acid (C12:0), myristic
acid (C14:0) palmitic acid (C16:0), stearic acid (C18:0), pal-
mitoleic acid (C16:1) oleic acid (C18:1) and linoleic acid
(C18:2). Overall, it was found that the biodiesel produced
from these cyanobacterial strains achieved the ASTM stand-
ard in terms of cetane number (51 to 68.94) and oxidation
stability (34.90 to 176.01 h).
Insect larvae
In recent years, many studies have reported on biodiesel
production using insect larvae as feedstock. In general,
insects are classified in the phylum Arthropoda and there
are about 1 million known species under the insect group
(Erwin 2004). Indeed, tropical countries are more favourable
for most insect species and contain more than 58% of the
global biodiversity (Foottit and Adler 2009). As discussed
by Manzano-Agugliaro et al. (2012), insect larvae have great
potential for nutritional and commercial due to their avail-
ability throughout the year. The life cycle of insects begins
with eggs and is constrained by an inelastic exoskeleton. The
life cycle starts with immature stages that are different from
the adult in habitat and structure, followed by a change to a
passive pupal stage that completes the metamorphosis (Li
et al. 2011a; Douglas 2007).
The attractive aspect of insect larvae is their ability to
biodegrade a number of organic wastes such as restaurant
kitchen waste, municipal solid waste and agricultural waste.
Insect larvae fed on organic wastes are able to absorb the
nutrients into their bodies, and 30% of lipids are stored in
insect larva as biomass (Čičková et al. 2015; Sheppard et al.
1994; Diener et al. 2011). There are four main criteria for
insect larva selection specifically for biodiesel production:
(1) fat content of the larvae, (2) speed of completion of the
life cycle, (3) reproductive capacity and (4) requirements for
space (Manzano-Agugliaro et al. 2012; Leong et al. 2015).
A number of studies have assessed the potential of insect
larvae as raw feedstock for biodiesel production. Several
species of insect larvae, such as Hermetia illucens (black
13
International Journal of Environmental Science and Technology
soldier fly, BSF) (Leong et al. 2015; Lin et al. 2011; Zheng
et al. 2012; Li et al. 2015), Musca domestica (house fly, HF)
(Yang and Liu 2014), Tenebrio molitor (yellow mealworm
beetle, YMB) (Zheng et al. 2013) and Chrysomya mega-
cephala (oriental latrine fly, OLF) (Yang and Liu 2014; Li
et al. 2012), have been evaluated for this purpose. Table 7
summarises the studies of insect larvae as feedstock for bio-
diesel production.
Li et al. (2011b) conducted a study on the optimum con-
version of selected organic wastes by BSF larvae for bio-
diesel production. They reported that of the organic wastes
studied, chicken manure produced the highest lipid yield of
32.8% followed by pig manure (20.4%) and cattle manure
(12.8%). Furthermore, biodiesel produced from chicken
manure showed the highest biodiesel yield of 93%. GCMS
analysis revealed that the FAME content of biodiesel derived
from BSF fed with chicken manure comprised lauric acid
(35.5%), oleinic acid (23.6%) and palmitic acid (14.8%). The
properties of biodiesel were reported as: density of 885 kg/
m3, viscosity of 5.8 mm2/s, flash point of 123 °C, ester con-
tent 97.2% and cetane number of 53. These biodiesel prop-
erties highlight the feasibility of BSF larvae for producing
biodiesel. The biodiesel produced was reported to contain
67.6% saturated fatty acids. It was reported that the higher
the saturated fatty acid content, the lower the tendency of
the biodiesel produced to be oxidised.
In another study, Li et al. (2011c) fed BSF larvae with
dairy manure for 21 days. They reported that the extraction
rate of crude lipid was up to 22.3%, with 96% yield of bio-
diesel. The biodiesel was then analysed, and the properties
of biodiesel were: density of 872 ± 0.3 kg/m3, ester content
of 97.2 ± 1.4%, viscosity of 4.5 ± 0.1 mm2/s, flash point of
121 ± 2.6 °C, water and sediment of 300 ± 3.7 mg/kg and
acid number of 0.8 ± 0.2 mg KOH/g. The fatty acid profiles
of BSF lipid were: capric acid (3.1 ± 0.03%), lauric acid
(35.6 ± 0.1%), myristic acid (7.6 ± 0.1%), pentadecanoic
acid (1.0 ± 0.06%), palmitoleic acid (3.8 ± 0.2%), palmitic
acid (14.8 ± 0.04%), oleinic acid (23.6 ± 0.3%), linolenic
acid (2.1 ± 0.3%), stearic acid (3.6 ± 0.1%) and nona-
decanoic acid (1.4 ± 0.02%). Despite the high saturated
fatty acid content of 58.2% which conferred great abil-
ity to withstand auto-oxidation, the acid number of the
biodiesel produced exceeded the European standard EN
14214. The high acid number may cause significant cor-
rosion to engines.
The optimum experimental condition for transesterifica-
tion of housefly (HF) larval oil was studied by Yang et al.
(2014). The extracted crude lipid was approximately 21.6%
and contained high amounts of FFA of 21.61 mg KOH/g,
which was then treated by transesterification. They con-
ducted transesterification study, and the optimum process
was achieved at a methanol to oil molar ratio of 8:1 with
2 vol.% of ­H2SO4 at a reaction temperature of 70 °C for 2 h,
International Journal of Environmental Science and Technology

Table 7  Biodiesel production from several insect larvae as feedstocks

Insect larvae Biomass Biodiesel properties Yield FAME composition, (%) References

Black soldier fly, Hermetia
illucens
Black soldier fly, Hermetia
illucens
Housefly, Musca domestica L. Swine manure
Yellow mealworm beetle,
Tenebrio molitor
Oriental Latrine fly,
Chrysomya megacephala
Black soldier fly, Hermetia
illucens
Black soldier fly, Hermetia
illucens
Chicken manure Density: 885 kg/m3 93%
Viscosity: 5.8 mm2/s
Flash point: 123 °C
Cetane number: 53
Dairy manure Density: 872 ± 0.3 kg/m3 96%
Viscosity: 4.5 ± 0.1 mm2/s
Flash point: 121 ± 2.6 °C
Water sediment:
300 ± 3.7 mg/kg
Acid number: 0.8 ± 0.2 mg
KOH/g
Density: 881 kg/m3 96.8%
Viscosity: 5.64 mm2/s
Flash point: 145 °C
Cetane number: 52
Decayed vegetables Density: 860 kg/m3 96.8%
Flash point: 127 °C
Cetane number: 58
Water content: 300 mg/kg
Methanol content: 0.2%
Pig manure Density: 0.89 g/cm3 96.6%
Viscosity: 5.1 mm2/s
Flash point: 138 °C
Cetane number: 56
Water content: 0.02 %
Acid value: 0.28 mg KOH/g
Fermented wheat Density: 872 kg/m3 96.18% Lauric acid: 58.5
bran Water content: 0.03 mg/kg
Ester content: 97.2%
Acid value: 1.1 mg KOH/g
Viscosity at 40 °C:
5.8 mm2/s
Flash point: 123 °C
Cetane index: 53
Fermented wheat Density: 875 kg/m3 94.14% NR
bran Water content: 0.03 mg/kg
Ester content: 98.3%
Acid value: < 0.8 mg KOH/g
Viscosity: 5.2 mm2/s
Flash point: 121 °C
Cetane index: 50
Lauric acid: 35.5 Li et al. (2011b)
Oleinic acid: 23.6
Palmitic acid: 14.8
Capric acid: 3.1 ± 0.03 Li et al. (2011c)
Lauric acid: 35.6 ± 0.1
Myristic acid: 7.6 ± 0.1
Pentadecanoic acid:
1.0 ± 0.06
Palmitoleic acid: 3.8 ± 0.2
Palmitic acid: 14.8 ± 0.04
Oleinic acid: 23.6 ± 0.3
Linolenic acid: 2.1 ± 0.3
Stearic acid: 3.6 ± 0.1
Palmitic acid: 29.1 Yang et al. (2014)
Oleic acid: 23.3
Palmitoleic acid: 17.4
Linoleic acid: 17.2
Linolenic acid: 19.7 Zheng et al. (2013)
Palmitic acid: 17.6
Linoleic acid: 16.3
Stearic acid: 11.4
Palmitic acid: 36.91 Yang and Liu (2014)
Oleic acid: 27.67
Palmitoleic acid: 10.89
Linoleic acid: 9.49
Nguyen et al. (2018)
Palmitic acid: 11.2
Myristic acid: 8.8
Oleic acid: 7.2
Nguyen et al. (2018)

NR not reported, FAME fatty acid methyl ester

obtaining 0.94 mg KOH/g of FFA. They treated the crude
lipid through alkaline transesterification using KOH as a cat-
alyst, which was reported to produce 19.5 g of biodiesel and
a conversion rate of 90.3%. The properties of biodiesel were
determined using standard procedures and were reported as:
density of 881 kg/m3, viscosity of 5.64 mm2/s, flash point
of 145 °C, ester content of 96.8% and cetane number of
52. Based on GCMS analysis, the biodiesel comprised
linoleic (17.2%), palmitoleic (17.4%), oleic (23.3%) and
palmitic (29.1%) acids. This result presents the feasibility
of HF reared on swine manure for biodiesel production. It
was noticed that the biodiesel produced was less resistant
to oxidation due to the high unsaturated fatty acid content
of 60.2%. Therefore, the authors suggested the addition of
oxidation inhibitors to improve the oxidation stability of the
biodiesel produced.
The potential of YMB larvae fed with decayed vegeta-
bles for biodiesel production was assessed by Zheng et al.
(2013). They successfully produced 0.8% w/w of biodiesel
from 17.7% of fat content. The biodiesel was analysed using

13

GCMS, and several fatty acids were determined, including
19.7% of linolenic acid, 17.6% of palmitic acid, 16.3% of
linoleic acid and 11.4% of stearic acid. The properties of the
biodiesel produced were: density of 860 kg/m3, ester content
of 96.8%, flash point of 127 °C, water content of 300 mg/kg,
cetane number of 58 and methanol content of 0.2%. How-
ever, it was noted that the viscosity and acid number were
above the permissible limit set by EN 14214 standard. It was
recommended to blend the YMB biodiesel with diesel to
reduce the viscosity. Meanwhile, the high acid number indi-
cated the high FFA content in the biodiesel. Therefore, it was
essential to set up an optimisation study of acid-catalysed
esterification able to reduce the FFA content.
Meanwhile, Yang and Liu (2014) conducted a study on
the feasibility of OLF larvae to degrade and convert pig
manure to biodiesel. They fed OLF larvae with fresh pig
manure that contained 73% moisture. They reported that
OLF larvae were able to reduce the pig manure biomass
from 700 kg to 18.2 kg. The OLF larvae oil was analysed by
using GCMS, and the main fatty acids detected were 36.91%
of palmitic acid, 27.67% of oleic acid, 10.89% of palmit-
oleic acid and 9.49% of linoleic acid. The overall quality of
biodiesel produced from OLF larvae fed with pig manure
was reported to be satisfactory and met EN 14214 standard.
The properties of biodiesel were: density of 0.89 g/cm3, vis-
cosity of 5.1 mm2/s, ester content of 96.6%, flash point of
138 °C, water content of 0.02%, cetane number of 56 and
acid value of 0.28 mg KOH/g. However, the viscosity and
oxidation stability of the biodiesel produced were out of the
permissible limits. In order to resolve these issues, it was
suggested to blend the OLF biodiesel with diesel to reduce
the viscosity and to add an antioxidant to increase oxidation
stability. An economic benefit study was also conducted,
which found that the bioconversion of 129 million tonnes
of swine manure by OLF larvae would produce up to 0.57
million tonnes of biodiesel.
The potential and optimal experimental condition for
biodiesel production using BSF larvae supported on an
immobilised enzyme, namely Novozym 435, were investi-
gated by Nguyen et al. (2018). They reported that the highest
biodiesel yield of 96.18% was obtained under conditions
of a methanol/fat molar ratio of 6.33, an enzyme applica-
tion rate of 20%, reaction temperature of 26 °C and reaction
time of 9.48 h. The properties of BSF larvae biodiesel were
reported to meet ASTM D6751 standard. The biodiesel pro-
duced had properties of density of 872 kg/m3, water content
of 0.03 mg/kg, acid value of 1.1 mg KOH/g, ester content
of 97.2%, viscosity at 40 °C of 5.8 mm2/s, flash point of
123 °C and cetane index of 53. The results from GCMS
analysis revealed that the main components of BSF biodiesel
were oleic acid (7.2%), myristic acid (8.8%), palmitic acid
(11.2%) and lauric acid (58.5%). Based on these biodiesel
13
International Journal of Environmental Science and Technology
properties, BSF larvae can be a capable biodiesel feedstock
when cooperating with a reusable enzyme, namely Novo-
zym 435. It was also noted that the high saturated fatty acid
content of BSF biodiesel, 84.7%, resulted in higher cetane
number and oxidative stability.
Another study by Nguyen et al. (2018) found that BSF lar-
val lipid has the feasibility to produce 94.14% biodiesel yield
in a direct transesterification by a combination of methanol
and a co-solvent. Direct transesterification was carried out
by mixing the ground dried BSF larvae with a solution (a
mixture of methanol, a co-solvent and sulphuric acid). It was
found that n-hexane was the most suitable co-solvent, giv-
ing the highest biodiesel yield of 63.37% compared to other
co-solvents, including petroleum ether (35.67%), chloroform
(48.50%) and acetone (54.83%). They also reported that the
optimum biodiesel yield was produced under conditions of
n-hexane/methanol volume ratio of 1:2 (v/v), a catalyst loading
of 1.2 mL, temperature of 120 °C and reaction time of 90 min.
The properties of the biodiesel produced met the standards
recommended by EN 14214, with density of 875 kg/m3, water
content of 0.03 mg/kg, acid value of < 0.8 mg KOH/g, ester
content of 98.3%, flash point of 121 °C, viscosity of 5.2 mm2/s
and cetane index of 50. This study revealed the potential of
direct transesterification for producing high yield of biodiesel
from BSF larvae. However, the authors also stressed that it
is a requirement to add a purification process to remove the
sulphur, since a high sulphur content (0.12%) was detected in
BSF larval biodiesel.
Conclusion
This review summarises the available biodiesel feedstocks:
vegetable oils, waste cooking oil, animal fats, microorgan-
isms and insect larvae, that have been developed and evalu-
ated for their potential application in biodiesel production. In
this review, we have focused on optimum conditions of trans-
esterification process for biodiesel production, the biodiesel
properties and FAME profiles for each available biodiesel
feedstock. It also discussed the application of heterogeneous
catalysts and immobilised enzymes to stimulate the transesteri-
fication process, as well as to produce high-quality biodiesel.
Great efforts have been made in recent years to identify new
biodiesel feedstocks and optimum conditions of transesterifi-
cation process to maximise biodiesel production. This review
provides an understanding and beneficial knowledge on the
great potential of biodiesel feedstocks for further development
of environmentally friendly and cheap biodiesel production
processes.
Acknowledgment S. Ishak thanks the Ministry of Education Malay-
sia for providing a MyBrain15 (MyPhD) Scholarship Award. The
authors also thank Mr Mah Iazam bin Mah Hussin from Eko Dinamik
Resources for technical assistance.
International Journal of Environmental Science and Technology

Compliance with ethical standards Borugadda VB, Goud VV (2012) Biodiesel production from renew-
able feedstocks: status and opportunities. Renew Sust Energ Rev
16:4763–4784
Conflict of interest The authors declare that they have no conflict of
Cai ZZ, Wang Y, Teng YL, Chong KM, Wang JW, Zhang JW, Yang
interest.
DP (2015) A two-step biodiesel production process from waste
cooking oil via recycling crude glycerol esterification catalyzed
by alkali catalyst. Fuel Process Technol 137:186–193
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