Biomass Conv. Bioref.
DOI 10.1007/s13399-014-0150-6
ORIGINAL ARTICLE
The potential of Reutealis trisperma seed as a new
non-edible source for biodiesel production
Holilah Holilah & Didik Prasetyoko & Titie Prapti Oetami & Eka Budiadi Santosa &
Yusuf Muhammad Zein & Hasliza Bahruji & Hamzah Fansuri &
Ratna Ediati & Juwari Juwari
Received: 12 June 2014 / Revised: 11 September 2014 / Accepted: 11 September 2014
# Springer-Verlag Berlin Heidelberg 2014
Abstract The potential of non-edible Reutealis trisperma oil
as a new carbon source for biodiesel synthesis was investigat-
ed. The oil content in the kernels by hot pressing method was
50–52 wt%. The oil was processed into biodiesel directly
without purification. The methanol concentration, reaction
temperature, and the amount of catalyst used are crucially
important to ensure high yield of biodiesel production. The
optimisation of the synthesis condition achieved 95.15 % of
biodiesel yield at 65 °C. The properties of biodiesel produced
such as density; 887 kg/m3, flash point; 148 °C, acid number;
0.41 mg KOH/g, iodine number; 28.43 g-I2/100 g, total glyc-
erol; 0.037 wt%, ester content; 97.18 wt%, pour point;
−15 °C, and cetane number; 63 meet with ASTM standard
D6751.
H. Holilah : D. Prasetyoko (*) : Y. M. Zein : H. Fansuri : R. Ediati
Laboratory of Materials Chemistry and Energy, Department of
Chemistry, Faculty of Mathematics and Natural Sciences, Institut
Teknologi Sepuluh Nopember, Surabaya, Indonesia
e-mail: didikp@chem.its.ac.id
D. Prasetyoko
e-mail: didik.prasetyoko@gmail.com
T. P. Oetami
PT Kemiri Sunan Drajat, Paciran, Lamongan, East Java, Indonesia
E. B. Santosa
PT Agrindo, Raya Driyorejo Km. 19, Gresik, East Java, Indonesia
H. Bahruji
Cardiff Catalysis Institute, School of Chemistry, Cardiff University,
CF10 3AT Cardiff, United Kingdom
J. Juwari
Department of Chemical Engineering, Faculty of Industrial
Technology, Institut Teknologi Sepuluh Nopember, Surabaya,
Indonesia
Keywords Reutealis trisperma seed . Non-edible oil .
Biodiesel . Properties
1 Introduction
The demand on energy is increasing at a substantial rate, and
currently the main source of energy is fossil fuel. The products
from fossil fuels combustion contribute to the environmental
problem such as non-renewable resources; fossil fuels are
estimated to be exhausted within 50 years [7]. Research to-
wards the development of alternative energy source has begun
with the focus that has been made on the production of fuel
from renewable resources, which are environmentally benign
and sustainable.
In the recent years, fuel that consists of mono-alkyl esters
of long-chain fatty acids known as biodiesel has been identi-
fied as ideal candidate for alternative fuel. Biodiesel fuel is
carbon neutral based on the fact that no harmful carbon
emission is released in the atmospheric environment during
combustion. In addition to eco-friendly, compared to fossil
fuels, biodiesel has many advantages such as it is produced
from renewable resources and biodegradable [12, 14, 28].
Studies have been focused on finding ideal renewable
sources for biodiesel production. Vegetable oils and animal
fats such as soybean oil (Glycine max), jatropha oil (Jatropha
curcas), rapeseed oil (Brassica napus), palm oil (Elaeis
guineensis), sunflower oil (Helianthus annuus), corn oil
(Zea mays), peanut oil (Arachis hypogaea), and cotton oil
(Gossypium spp.) [20] were used as the carbon sources.
Moreover, hazelnuts oil (Corylus spp.) [29], yellow horn oil
(Xanthoceras sorbifolia Bunge.) [18], Phoenix dactylifera,
edible oil from Phoenix genus [2], and Ceiba pentandra [22]
have been used for biodiesel production. The main disadvan-
tage concerning about biodiesel production from these feed-
stocks is the high price of combustible vegetable oils
 Biomass Conv. Bioref.

compared to that of fossil-based diesel fuel. Zhang et al. [33]
reported that the cost of vegetable oils contributes to almost
70–95 % of the total cost of biodiesel production. The cost can
be reduced by using non-edible oil and waste oil as feed-
stocks. Recently, C. pentandra [22], Silybum marianum L.
[8], Datura stramonium L. [26], and Cerberra odollam [15] as
non-edible oils have been used for biodiesel production. Here,
we study the potential of Reutealis trisperma oil as a new non-
edible carbon source for biodiesel production. In this study,
R. trisperma oil was converted into biodiesel through esteri-
fication and transesterification reactions using H2SO4 and
NaOH as catalysts. The seed of R. trisperma is shown in
Fig. 1.
2 Materials and methods
2.1 Materials
R. trisperma oil used in this study was supplied by PT Kemiri
Sunan Drajat (Lamongan, Indonesia). The oil content in the
kernels by hot pressing method was 50–52 wt%. The oil was
processed into biodiesel directly without purification.
Methanol (99%) was purchased from Merck (Darmstadt,
Germany). Sulfuric acid (H2SO4; 98 %) and sodium hydrox-
ide (NaOH; 99 %) as catalysts were purchased from Merck
(Darmstadt, Germany). The standards of methyl esters of fatty
acids including methyl palmitate (C16:0), methyl stea-
rate (C18:0), and methyl oleate (C18:1) (99 %) were
purchased from Merck (Darmstadt, Germany). Methyl
heptadecanoate (99 %, Merck) was used as an internal
standard, while n-hexane (99 %, Merck) was used as an
analytical solvent.
2.2 Methods
2.2.1 Esterification of Reutealis trisperma oil
Esterification of high free fatty acid content in R. trisperma oil
with methanol was carried out using H2SO4 as catalyst with
1:1 to 1:3 methanol to oil weight ratio was used. The reaction
was carried out in a round bottom flask equipped with a reflux
condenser and a magnetic stirrer. Methanol and 3.0 wt%
H2SO4 were added into the round flask containing the oil,
and the mixture was vigorously stirred. The mixture was
heated at 65 °C in oil bath for 2 h. Two layers of product were
formed, and the mixture was separated using separating fun-
nel. The top layer was the residual containing methanol and
water, while the bottom layer was the product of esterification
reaction that will later be used for the subsequent
transesterification reaction.
2.2.2 Transesterification of Reutealis trisperma oil
The product obtained from esterification, esterified R. trisperma
oil (RTOE), was further used in transesterification reaction with
methanol to produce biodiesel. The reaction was carried using
1.0 wt% NaOH base catalysts with 1:2 weight ratio between

Fig. 1 Seed of Reutealis

trisperma
Biomass Conv. Bioref.
RTOE and methanol. The mixture was added into a round
bottom flask equipped with condenser. The reaction was carried
in an oil bath at 65 °C for 1 h under a constant stirring speed [29].
The round bottom flask was immersed in the water bath until two
layers of product formed; the top layer was mainly methyl ester
and the bottom layer was mainly glycerol. The glycerol was
removed using a separating funnel while the obtained methyl
ester was washed with hot distilled water and ethanol until a clear
solution was formed. In order to investigate the optimum reaction
condition for the reaction, the amount of catalysts used were
varied from 0.5, 1.5, and 2.0 wt%. The effect of reaction tem-
perature on the rate of biodiesel production was also investigated
at 30, 50, and 70 °C. We also varied the weight ratio of methanol
to oil from 1:1 to 1:3.
2.2.3 Analysis of biodiesel properties
The composition of methyl esters in the obtained biodiesel was
analyzed using a gas chromatograph with MS detector (GC, HP
6890) and gas chromatography with FID detector (GC,
Techcomp 7900). The GC was equipped with a non-polar
ECTM-5 capillary column containing 5 % phenyl and 95 %
methyl polysiloxane (30 m in length, 0.25-mm internal diameter,
and 0.25-μm film thicknesses). The column temperature was set
at initial temperature of 200 °C (2 min) with heating rate of
5 °C/min to 220 °C (5 min) and 4 °C/min to 250 °C (1 min).
Both of the injector and the detector temperatures were set at
250 °C. Seventy milligrams of biodiesel was dissolved in 1-mL
n-hexane and then injected in the GC using a microliter syringe.
The measurement was run in 17 min using methyl
heptadecanoate as an internal standard. The other properties of
biodiesel also has been analysed based on cetane number,
kinematic viscosity, condradson carbon residue, density, flash
point, pour point, cloud point, and acid number according to
standard methods. The following equation reported by Yang
et al. [31] was used to calculate the yield of biodiesel:
=CI
WB
Yield ¼
WA  C  100 %
Where WB is the weight of produced biodiesel (g), CI is the
weight of injected biodiesel into GC (ppm), WA is the weight
of the oil (g), and C is the concentration of methyl ester in the
biodiesel (ppm).
3 Results and discussion
3.1 Esterification of Reutealis trisperma oil
The presence of high-content free fatty acid (FFA) in vegeta-
ble oil affects the conversion to biodiesel due to saponification
reaction with the base catalysts to produce soap [3, 34]. The
FFA content in R. trisperma oil was determined to be around
2.4 % using NaOH titration. To prevent a competing saponi-
fication reaction, the oil must undergo esterification reaction
to reduce the FFA content. The amount of FFA in vegetable oil
should be less than 1.0 wt% to ensure a good conversion to
biodiesel [19]. The esterification pretreatment on the
R. trisperma oil was successfully reduced the high FFA con-
tent to only 0.09 % with 96.3 % of FFA was converted using
sulfuric acid catalyst. This is in agreement with the observa-
tions by Marchetti and Errazu [19] that shows 96 % of FFA
conversion on the esterification of sunflower oil using sulfuric
acid as a catalyst.
3.2 Transesterification of Reutealis trisperma oil
Transesterification of R. trisperma oil containing 0.09 % FFA
was carried out with methanol using NaOH as a base catalyst.
Analysis using gas chromatograph reveals the resultant bio-
diesel contains several major types of methyl esters such as
methyl palmitate, stearate, and oleate. Methyl esters profile of
R. trisperma biodiesel are shown in Table 1. The concentra-
tion of each methyl ester was determined based on the peak
area of the product obtained from gas chromatograph. The
yield of biodiesel was then calculated using the formula
reported by Yang et al. [31] and the results given in Table 2.
3.3 Effect of catalyst amount
There is a direct correlation between the amount of catalyst and
the yield of biodiesel production [25, 10]. In this study, we varied
the amount of catalyst from 0.5, 1.0, 1.5, and 2.0 wt% in order to
get the optimum value for the reaction. Results given in Table 2
show that the optimum catalyst for the reaction is 1 wt% with the
yield of biodiesel is 85.1 %. Significant improvement in the
amount of biodiesel yield from 50.65 to 85.16 % was observed
when the catalyst used in the reaction was increased from 0.5 to
1.0 wt%. However, further increased of the amount of the
catalyst to up to 2 wt% leads to the formation of soap via
saponification process and this significantly reduced the
Table 1 Methyl esters profile of Reutealis trisperma biodiesel
Type of methyl ester in biodiesel composition Content (wt%)
Methyl palmitate 22.92
Methyl palmitoleate 0.33
Methyl stearate 21.95
Methyl oleate 30.16
Methyl linoleate 13.60
Methyl linolenate 1.80
Methyl arachidate 0.31
Methyl eicosenoate 0.79
 Biomass Conv. Bioref.

Table 2 Biodiesel yield on the transesterification of esterified Reutealis 3.3.1 Effect of methanol to oil weight ratio
trisperma oil for 1 h

Catalyst amount Methanol to oil Temperature Yield The ratios of methanol to oil used in the reaction were varied
(wt%) weight ratio (°C) (%) from 1:1, 1:2, and 1:3 weight ratios. The amount of biodiesel
produced for each reaction was given in Table 2. Clearly, the
0.5 1:2 65 50.65
high ratio of methanol to oil used in the reaction enhances the
1.0 1:2 65 85.16
production of biodiesel. When the reaction was carried out
1.5 1:2 65 68.04
using the optimum catalyst concentration of 1.0 wt% and a
2.0 1:2 65 40.77
temperature of 65 °C [9], 95.15 % of biodiesel yield was
1.0 1:1 65 95.15
produced at a methanol to oil weight ratio of 1:1. It is known
1.0 1:3 65 50.09
that the biodiesel production via transesterification is a revers-
1.0 1:1 30 32.61 ible reaction; the presence of excess alcohol is the system
1.0 1:1 50 65.55 inhibits the equilibrium and favors backward reactions [7].
1.0 1:1 70 75.03 Dehkordi and Ghasemi [6] also found a significant improve-
ment in biodiesel yield with the increasing of methanol to oil
weight ratio. However, there is a limitation where at a certain
production of biodiesel [13]. Optimum amount of catalysts is point, the amount of excess methanol has no significant effect
important to drive the reaction as a lack of catalyst may cause on the biodiesel yield produced. At high methanol concentra-
incomplete conversion of reactants [27, 30]. Agarwal et al. [1] tions, product purification involving separation from metha-
found similar observation on the transesterification of waste nol may take a long time to complete [30].
cooking oil using KOH as catalyst, which the methyl ester yield
increases to 98.7 % with the catalyst amount and further increase
of the catalyst decrease the methyl ester yield. Due to the 3.3.2 Effect of reaction temperature
formation of soap, the further separation was needed to separate
the soap with the methyl ester. Besides the amount of catalyst and weight ratio of methanol to
The amount of the catalyst used during the reaction repre- oil, temperature also has an important role in the
sents the number of active sites available for the reaction to transesterification reaction. From Table 1, the reaction tem-
occur and to relate the turnover frequency (TOF) parameter perature was changed from 30 to 70 °C, and the optimum
used. TOF is defined as the mole of product (yield of biodie- temperature for the reaction is at 65 °C where 95.15 % of
sel) for every mole of the active site on the catalysts over the biodiesel has been produced. The maximum yield was obtain-
reaction time. The correlation between the number of active ed at 65 °C reaction temperature since this temperature was
site, the yield of biodiesel produced, and the TOF are given in the boiling point of methanol, so that the temperature was the
Table 3. The TOF increases with the amount of the catalyst most suitable for the transesterification reaction. When the
used in the reaction, but reduced significantly at 1.5 and reaction temperature is higher than the boiling point of meth-
2.0 wt%. At optimum catalyst amount of 1 wt%, the higher anol, the bubbles are formed on the surface inhibiting the mass
TOF of 34.50 h was obtained. The high TOF showed the high transfer phase as described by Tang et al. [24] and Zabeti et al.
activity of the catalyst due to its high amount of active site [32]. The higher temperature used, the faster rate of the
resulting in high biodiesel yield. But, there is a limitation for reaction, is achieved for the endothermic reaction causing
transesterification using NaOH as a catalyst because its excess the higher yield. However, there is an optimum temperature
amount leads to the formation of soap through saponification used for the reaction in this study related to the boiling point of
reaction lowering the biodiesel yield [29]. methanol [18]. Xu and Hanna [29] observed the
transesterification of hazelnut oil (Corylus spp.) using NaOH
as a catalyst resulting the maximum yield at a temperature of
Table 3 Correlation of active site, yield, and TOF. Reaction conditions:
methanol to oil weight ratio, 1:2; temperature, 65 °C
65 °C. Since methanol’s boiling point is 65 °C, at this tem-
perature, it can diffuse completely to react with triglyceride
Catalyst amount Active site Biodiesel yield TOF molecules.
(wt%) (mol)a (%) (h)

0.5 0.00375 50.65 16.00

1.0 0.0075 85.16 34.50 3.4 Properties of biodiesel produced
1.5 0.01125 68.04 19.80
2.0 0.015 40.77 11.30
We used biodiesel previously produced from S. marianum L.
to investigate the properties of biodiesel produced from
a
Active site is the mole number of the catalyst R. trisperma oil. The data were given in Table 4.
Biomass Conv. Bioref.

Table 4 Properties of biodiesel produced from Reutealis trisperma oil

Properties Method Unit ASTM D6751 Reutealis Silybum

(limit) trisperma marianum La

Density at 15 °C ASTM D-1298 kg/m3 860–900 887 878

Viscosity kinematic at 40 °C ASTM D-445 mm2/s (cSt) 3.5–5.0 6.71 4.61
Flash point ASTM D-93 °C 101 min 148 85
Acid number ASTM D-664 mg KOH/g 0.5 0.41 0.08
Iodine number AOCS Cd 1-25 g-I2/100 g 120 max 28.43 99
Calorie Bomb calorimeter cal/g – 9208 9673
Total glycerol AOCS Ca-14-56 %-m 0.25 max 0.037 –
Ester content Calculated %-m 96.5 min 97.18 95
CCR ASTM D-4530 % m/m 0.05 max 0.25 0.02
Pour Point ASTM D-97 °C 18 max −15 −12
Cetane Number ASTM D-613 – 51 min 63.5 43.01
Cloud Point ASTM D-2500 – – −13 −2
a
Adapted from Fadhil et al. [8]

3.4.1 Cetane number the high cetane number than the biodiesel produced from
other non-edible oils.
Cetane number is an index that is commonly used to deter-
mine the quality of fuel especially during combustion. It gives 3.4.2 Kinematic viscosity
an indication towards the combustion quality of the biodiesel
during compression ignition. The combustion quality of the Kinematic viscosity is important features of the fuel that
biodiesel is closely related to the structure of the fatty acid influences the fuel spray, the mixture formation, and the
methyl ester components [16]. Data given in Table 4 shows combustion process. Fuel with high viscosity inhibits the
that the cetane number of biodiesel produced from injection process and leads to inadequate fuel atomization.
R. trisperma oil was 63.5 relatively higher than the cetane The average diameter of the fuel driblets from the injector
number of biodiesel produced from S. marianum L, 43.01. We and their permeation increases with the increasing of fuel
also found that the cetane number of biodiesel produced in this viscosity [5]. As given in Table 4, we found that the kinematic
study is significantly higher from non-edible oils such as viscosity of biodiesel produced from R. trisperma oil was
J. curcas L. (57.1) and Xanthium sibiricum (48.8) [21, 4]. 6.715 mm2/s which is relatively higher than the kinematic
R. trisperma oil produced a good quality biodiesel based on viscosity of biodiesel derived from S. marianum L.,
4.61 mm2/s [8]. This result, however, is above the standard
ASTM D445 value which is within the range of 1.9–6.0 mm2/
Table 5 The acid value comparison of Reutealis trisperma seed oil with s. We believe the presence of triglycerides produced as a by-
other oils for biodiesel production product from the unreacted R. trisperma oil affects the viscos-
ity of the biodiesels. We also observed that the kinematic
Oil Acid value (mg KOH/g)
viscosity of the as-synthesis biodiesel derived from
Reutealis trisperma seed 11 R. trisperma oil is higher than the other biodiesel from non-
Soybeana 0.2 edible oils such as J. curcas L and C. pentandra [23, 22].
Rapeseeda 2.93
Sunflowera 0.15 3.4.3 Condradson carbon residue (CCR)
Palma 0.1
Peanuta 3 Condradson carbon residue (CCR) indicates the amount of
Corna 0.11 carbon that not converted into energy during the combustion.
Jatropha curcasa 28 The CCR of the produced biodiesel was measured around
Palangaa 44 0.254 % m/m (Table 4). This result was not in accordance
Sea mangoa 20 with the ASTM D4530 standard (max 0.05 % m/m). The CCR
of biodiesel produced from R. trisperma oil was the highest
a
Adapted from Leung et al. [17] than that of from S. marianum L. (0.03 % m/m) and

C. pentandra oil (0.02 % m/m) [21]. The high CCR was
achieved since the produced biodiesel still contain most tri-
glycerides derived from unreacted R. trisperma oil affecting
the biodiesel carbon residue.
3.4.4 Evaluation the other properties of biodiesel produced
Besides cetana number, kinematic viscosity, and CCR, other
properties of the produced biodiesel were also determined.
Based on data given in Table 4, it can be concluded that the
properties of biodiesel based on density, flash point, pour
point, cloud point, and acid number were in accordance with
the ASTM standards. Biodiesel produced from R. trisperma
and S. marianum L. shows different value in iodine number.
Iodine number is used to indicate the degree of unsaturated
fats and oils and in biodiesel, iodine number is used to com-
pare the number of C=C content in natural complex mixture
of fatty acid alkyl esters [11]. Iodine number of the biodiesel
produced from R. trisperma and S. marianum L. was 28.43
and 99 g-I2/100 g, respectively, indicating the amount of
double bond in the fatty acid methyl esters obtained from
R. trisperma oil was less than that of from S. marianum L.
4 Conclusion
R. trisperma oil was investigated as raw material for biodiesel
production via esterification and transesterification reactions with
methanol (Table 5). Pretreatment of R. trisperma oil via esterifi-
cation with using H2SO4 catalysts reduced the free fatty acid
level and prevents saponification process. Transesterification
reaction was optimised by varying the amount of catalyst, the
weight ratio of methanol to oil, and the reaction temperature. The
optimum reaction condition for biodiesel production with the
maximum yield of 95.15 % was obtained at 65 °C, with meth-
anol to oil weight ratio of 1:1 and the amount of catalyst used is
1.0 wt%. The properties of produced biodiesel were tested, and
most properties were in accordance with ASTMD 6751-10
standard, except kinematic viscosity and condradson carbon
residue (CCR).
Acknowledgments The authors acknowledge the research grant pro-
vided by Ministry of Education and Culture, Indonesia, under
“Unggulan” research grant EPI-UNET-2013 and Japan International Cor-
poration Agency (JICA) B1-1, 2012–2013.
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