Journal of the Japan Institute of Energy, 94, 763-768（2015） 763

Production of Biodiesel from Rubber Seeds (Hevea Brasiliensis)

by In situ Transesterification Method

Bashir Abubakar ABDULKADIR ※ 1, Yoshimitsu UEMURA ※ 2, Anita RAMLI※ 1, Noridah Bt OSMAN※ 2,

Katsuki KUSAKABE ※ 3, and Takami K AI ※ 4
(Received April 4, 2014)

Biodiesel is defined as the mono-alkyl esters of vegetable oils (both edible and non edible) or animal fats, obtained
by transesterifying oil or fat with an alcohol. In this research, biodiesel is produced by in situ transesterification
(direct transesterification) method from the seed of rubber tree using KOH as a catalyst. The influence of seed to
methanol mass ratio, reaction time, temperature, seeds sizes, and catalyst loading was investigated. Important properties of
biodiesel such as free fatty acid (FFA), viscosity, pour point, density etc were also investigated. The result showed that
the best ratio of seeds to methanol was 1:6 (10 g seeds with 60 g methanol), 120 min reaction time at 60 °C reaction
temperature, and catalyst loading of 3.0 g with 150 µm seeds size. The maximum yield obtained was 96 %, and also most of
the properties obtained were in agreement with the standards and with other FAMEs. This study supports the production of
biodiesel from the rubber seeds using in situ (direct) transesterifcation method from the seeds as an alternative to diesel
fuel.

Key Words
Biodiesel, FAME, Free fatty acid, in situ

in the diesel engine and in the existing fuel distribution and

1. Introduction storage infrastructure. Moreover, biodiesel has a high energetic
According to the Brazilian biodiesel programme, yield, fixes the solar energy and contains insignificant
biodiesel is defined as a fuel obtained from mixtures, in amounts of sulphur. Therefore, biodiesel is currently the best
different proportions of fossil diesel and alkyl esters of substitute for fossil diesel 1) 2).
vegetable oils or animal fats 1). However, under the chemical point In recent discussions of biodiesel production by
of view, biodiesel is composed mainly of fatty acids mono-alkyl transesterification process, a controversial issue has been
esters. Transesterification is a process used for obtaining whether biodiesel can be produced by in-situ (direct)
biodiesel and involves the catalyzed reaction of triglycerides transesterification process. Widayat et al. 3), reported that,
(major compounds of oils and fats) with short-chain alcohols biodiesel production from either vegetable oil or animal fat has a
such as methanol and ethanol. Most transesterification in major drawback. The essence of Widaya’s argument is that to
industrial processes employ alkaline catalyst (KOH, NaOH, and obtain the vegetable oil by conventional method, the seed is
NaOCH3) and methanol 1). Its use (pure or blended) does not pressed by mechanical process or extraction with chemical
demand any modification solvent. The oil is further pretreated by degumming process. Of
course this process requires a lot of manufacturing cost. The
※ 1 Department of Fundamental and Applied Sciences, biodiesel production from rubber seed oil used two stage-
Universiti Teknologi PETRONAS
3150 Tronoh,Malaysia method of esterification reaction followed by transesterification
※ 2 Centre for Biofuel and Biochemical Research (CBBR), reaction 4) 5). This process still has the same drawbacks that are
Universiti Teknologi PETRONAS the pretreatment process of the oil. Most studies in the field
Tronoh, Malaysia of biodiesel production have only focused on vegetables oil and
※ 3 Department of Nanoscience, Faculty of Engineering, Sojo animal fat. Therefore, to solve these drawbacks, the
University (trans)
4-22-1 Ikeda, Kumamoto 8600082, Japan
※ 4 Department of Chemical Engineering, Faculty of
Engineering, Kagoshima University 1-21-40
Korimoto, Kagoshima 890-0065, Japan
 764 J. Jpn. Inst. Energy, Vol. 94, No. 7, 2015

esterification process by in situ method will be one of the best

possible alternatives. weighing 2-4 g each. The capsules of the seeds are spread over
the ground in plantation. The seeds were collected from a
In-situ (trans)esterification is a method used to plantation nearby Universiti Teknologi PETRONAS, Perak
produce biodiesel where the raw material (seed) is contacted Malaysia and dried under the sun for about 3 h to remove
directly with an alcohol assisted with an acid or alkaline the remaining moisture, then, the shell and kernels were
catalyst. Thus extraction and transesterification take place in separated by breaking the capsules. The kernels were dried in
one step and the alcohol is used for extracting oil from the the oven for about 24 h to remove the remaining moisture. The
seeds as well as transesterifying it 6). In this process, mass ratio kernels were immediately ground using food processing
of seeds to methanol must be higher than the value calculated machine ( blender) in order to weaken and rupture the cell.
based on the stoichiometry. Excess methanol will play dual Subsequently, the ground rubber seed was taken into the oven
role as the solvent extraction as well as the reactant 7). The cost and kept overnight at 104 °C in order to deactivate the remaining
of biodiesel production process could be minimized if in situ enzyme as well as to remove the moisture and then sieved using
(trans) esterification method is used. In this process, costs for 300, 350, and 400 µm opening screens. Finally, three different
solvent extraction and oil purification can be reduced. So, size fractions were obtained; 150 µm (0-300 µm), 325 µm
biodiesel production becomes easier and simpler 8). In-situ (300-350 µm), and 375 µm (350-400 µm). After drying, the
transesterification of sunflower oil with acidified methanol moisture content of the kernels was determined using halogen
produces fatty acid methyl esters in yield significantly greater moisture content analyzer (Mettler Toledo, HR83) and also after
than those obtained from conventional process 3). the water is removed from the seeds by evaporation using
In-situ transesterification was first introduced by furnace, and in both cases it was found to be 1.5 %. The oil
Harrington and D’Arcy-Evans 7). They used sunflower seed as content of the kernels was determined through soxhlet
a raw material. Marinkovic et al. 9), has done experiments using extraction using hexane as a solvent for 4-5 h. It was found that,
sunflower seed with the same process. Harrington et al. 7), have the seed has oil content of 40 %.
done in-situ process from soybeans. Ozgul-Yucel and Turkay 10)
11)
performed in situ esterification process with rice bran as raw 2.2 Rubber seeds characterization
material. Also Ginting et al. 12), performed in situ
transesterification from castor seed. Ozgul and Turkay 13) The recommended methods of the Association of
investigated in situ esterification of high-acidity rice bran oil Official Analytical Chemists (AOAC, 1984) were adopted to
with methanol and ethanol, and with sulfuric acid as catalyst. determine the levels of ash and water content from the seeds.
The transesterification with in situ method gave similar ester
yields to those obtained from conventional transesterification, 2.3 Transterification process
being an alternative, efficient, and economical process 13). Methanol and the KOH catalyst were mixed in a
The objectives of this research are to produce three neck round bottom flask connected to a reflux condenser
biodiesel from rubber seed by in-situ transesterification method and heated until catalyst completely dissolved. Then, 10 g of the
and to investigate the influence of reaction parameters on the ground rubber seeds sample was added into the mixture and the
FAME yield. The merit of this method is that this new blend was heated at 60 °C and at atmospheric pressure for 120 min
process just requires a single process, where the normal oil at 500 rpm stirring speed. The product was filtered using
extraction process can be avoided. In addition, in this process, Whatman filter paper and poured into the separating funnel to
the purification of products after transesterification reaction is separate the excess of alcohol. The reaction was repeated to
much simple compared to the common method. Therefore, this study the effect of rubber seeds to methanol mass ratio, reaction
new process can offer an alternative way to convert the seeds temperature, reaction period, catalyst loading, and sizes of the
directly to methyl esters by a simpler-shorter production seeds on the FAME yield. The FAMEs analysis was quantified
process. by Agilent Technologies 7890A Gas Chromatography equipped
with Flame Ionization Detector (GC-FID) with HP-5 5%
Phenyl Methyl Siloxane capillary column (30 m by 320 µm by
2. Experimental Procedure 0.25 mm). Automatic injection sampler by Agilent was used.
2.1 Sample collection and preparation Methyl heptadecanoate (0.630 mg internal standard) was
dissolved in 1 mL hexane to prepare the standard
Rubber trees yield 3-seeded ellipsoidal capsules, each
capsule with three seeds. Rubber seeds are ellipsoidal, variable
in size, 2.5-3 cm long, mottled brown, lustrous,
 J. Jpn. Inst. Energy, Vol. 94, No. 7,
765
2015

solution. Approximately 100 mg crude methyl ester was

dissolved in 1 mL standard solution for GC-FID analysis. Table 1 Characteristics of rubber seed/rubber seed oil
About 1 µL sample was injected into the GC at an oven
temperature of 210 °C with helium as the carrier gas. The GC oven Component Content (wt %)
Seed
was programmed at 210 °C, isothermally for 15 min. Then, the
FAMEs content and the FAME yield were calculated. The Moisture 1.50
experimental design and response/FAME conversion conducted Ash 2.88
in this research paper is similar to a conventional method of Oil 40.00
FAME production from rubber seed oil by Junaid et al. 14). Seed oil
The FAME yield was calculated using the following
expressions 15): Water 3.30
Moil phase×XFAME
FAME Yield = ×100 % similar with that of Widayat and Suherman 19).
Mseed×Xoil×F
Moil phase = Mass of oil phase after transeterification (g). XFAME 3.2 Effect of seed to alcohol mass ratio
= Mass fraction of FAME in oil phase. The mass ratio of seeds to alcohol is one of the
Mseed = Mass of the seeds (g). important factors that affect the FAME yield as well as
production cost of biodiesel. Mass ratio is the ratio of weight
Xoil = Mass fraction of oil in seeds. of seeds to the weight of alcohol. The FAME yield is defined
F = 3MWFAME/MWoil (MW= molecular weight); as the yield of the process represented in terms of percentage 20).
approximately unity. In this research, various mass ratios of the seeds to alcohol
(methanol) starting with 1:2 and followed by (1:3, 1:4, 1:5, 1:6,
1:7 and 1:8) were tested. It was found that, the maximum yield
3. Result and Discussion was achieved at the mass ratio of 1:6 (96 %) as shown in Fig. 1
3.1 Rubber seeds characterization and further increase in mass ratio, there was no any improvement
Water content or moisture content is the quantity of in the conversion efficiency, instead a decrease was observed.
water contained in a material, such as in soil (called soil
moisture), oil, fruit, seeds or wood. Water content is used in a 3.3 Effect of reaction temperature
wide range of scientific and technical areas, and is expressed as The FAME yield was only 67 % at 40 °C, but increased
a ratio, which can range from 0 (completely dry) to the value of rapidly with increasing in temperature. The optimum
the materials' porosity at saturation 16). The moisture content in temperature for this reaction was found to be at 60 °C, with 96
this paper was determined by measuring the mass of rubber % FAME yield with 1:6 mass ratio, 3 g KOH catalyst and 120
seeds before and after the water is removed from the seeds by min reaction time as shown in Fig. 2.
evaporation using furnace while the water content for the oil The reason for the increase in yield as the temperature
was determined using Karl Fischer titration process based on increases is that, increase in temperature is believed to facilitate
AOCS standard. the breaking of the seeds cell walls, thereby creating a void
For many samples, the water content may be an which serves as migratory space
extremely important index used for establishing the
relationship between the samples/rubber seed oil and its
properties. The uniformity of a fine-grained seeds largely
depends on its moisture content 17). In this research, a
Malaysian rubber seeds was found with 1.5 % moisture
content while rubber seed oil was found with less than 3.3 %
water content as shown in Table 1.
Ash content is the name given to all non-aqueous residue
that remains after a sample is burned, which consists mostly of
metal oxides. Ashing is the process of mineralization for pre-
concentration of trace substances prior to chemical analysis 18).
In this research, Malaysian rubber seeds was found to have 2.88
% ash content as shown in Table 1, and the result was found
to be very

Fig. 1 Effects of mass ratio on the FAME yield (%) (temp= 60

°C, seeds size= 150 µm, time = 120 min)
766 J. Jpn. Inst. Energy, Vol. 94, No. 7, 2015

Fig. 2 Effects of reaction temperature on the FAME yield (%)

(seeds to MeOH mass ratio= 1:6, seed size= 150 μm, time
= 120 min)

for the contents of the oil bearing cells. Temperature also

lowers the viscosity of the oil, thus facilitating the release of the
oil out of the kernel. However, at higher temperature, there is
substantial loss of moisture leading to a hardening of samples
and oil degradation. This may account for the decrease in the
yield at higher temperature of 65 °C 21).
As said by Ramadhas et al. 4) reaction temperature greater
than 60 °C should be avoided in the case of rubber seed, because
they tend to accelerate saponification before transesterification
reaction completed. Fig. 3 Effects of catalyst concentration on the FAME yield (%)
(Seeds to MeOH mass ratio= 1:6, seed size= 150 µm, 60 °C,
3.4 Effect of catalyst concentration. 120 min)
The KOH concentration of 1.0-4.0 g was used for the
analysis. The maximum FAME yield of 96 % was
achieved at 3.0 g KOH loading with 1:6 mass ratio of seeds to
methanol, 120 min reaction time, and at 60 °C reaction
temperature, as shown in Fig. 3. It was reported 22) that, in- situ
transesterification is inactive without catalyst. Acid or alkali
catalyst in in-situ transesterification helps to break the cell
wall of the seeds, thereby facilitating methanol to access the
oil in cotyledon cells.
As mentioned by Ramadhs et al. 23) from physical
appearance, the higher the catalyst concentration, the darker
biodiesel will be produced. Therefore the addition of a suitable
catalyst concentration is important to the physical appearance of
biodiesel.

3.5 Effect of reaction duration

The maximum FAME yield obtained in this
experiment (effect of reaction duration) was 96 % at 120 min
reaction duration with 1:6 mass ratio of seeds to methanol, 3 g KOH Fig. 4 Effects of reaction time on the FAME yield (%) (seeds to
catalyst loading, and at 60 °C reaction temperature as shown in MeOH mass ratio= 1:6, temp = 60 °C, seed size= 150
Fig. 4.
µm, time = 120 min)

Fig. 5 Effect of mesh size (µm) on ester content (%) (seeds to

MeOH mass ratio= 1:6, temp = 60 °C, time = 120 min)

3.6 Effect of sizes of the seed.

Various sizes of the seeds ranging from 300 to 400
µm in size were used. It has been observed that, the maximum
yield of 96 % was achieved at the smallest mesh sizes of 150
µm as shown in Fig. 5.
This phenomenon could be attributed to the fact that
smaller particles have larger amount of surface area as well
 J. Jpn. Inst. Energy, Vol. 94, No. 7,
767
2015

as an increased number of broken cells resulting in a high oil

concentration at the smaller particle sizes 21). atomization of the fuel spray and less accurate operation of the
The FAME yield obtained in all the optimum fuel injectors 26). In this study the viscosity obtained was 4.68 mPa
conditions was 96 % as discussed. This is one of the s as shown in Table 2 which falls within the range of ASTM
observed drawbacks of in-situ transesterification process that it standard. This shows that FAME produced through an in situ
can not go beyond 96 % based on this paper and other previous process will serve as an excellent alternatives to the energy
work, and this might be due to the extraction and crisis since it has low viscosity.
transesterification taking place at the same time, and also single Density is an important property of biodiesel. Density is
solvent (methanol) used for extracting the oil as well as the mass per unit volume of any liquid at a given temperature
transesterifying it unlike conventional method of while specific gravity is the ratio of the density of a liquid to the
transesterification. But further studies may improve the yield density of water. Density and specific gravity has importance in
beyond the acquired result in this paper. diesel-engine performance since fuel injection operates on a
volume metering system 25). Also, the density of the liquid
product is required for the estimation of the Cetane index 27).
4. Properties of the Produced FAME Two important parameters for low-temperature
The fuel properties of rubber seed methyl ester in applications of a fuel are cloud point (CP) and pour point (PP). The
relationship with those of other esters are shown in Table 2. The CP is the temperature at which a cloud of crystals first appears
majority of the properties of the obtained FAME through in in a liquid when cooled under conditions as described in
situ method are relatively equivalent to those of other esters. ASTM D2500-91. The PP is the temperature at which the
The GC-FID analysis shows that, the biodiesel contains amount of wax from solution is sufficient to gel the fuel; thus
large amount of C18. The present result shows that, in situ it is the lowest temperature at which the fuel can flow. The
transesterification process enhanced the fuel properties of the PP is the lowest temperature at which an oil specimen can still
oil with respect to specific gravity, viscosity, density, and acid be moved. It is determined according to ASTM D97-96. These
value. The viscosity of biodiesel is in the range of biodiesel two properties are used to specify the cold-temperature usability of
standard. The properties of the obtained biodiesel are in a fuel 28). In this experiment, both the pour point and cloud point
comparable with ASTM standards. The observed properties of fall within the ASTM standard described in Table 2. Also, it has
methyl esters from rubber seed by in situ transesterification been observed in this study that, the acid value was less than
methood are found to be in reasonable conformity with 1 mg-KOH/g which is within the acceptable limit of ASTM.
ASTM 6751.
The present results obtained show that, the 5. Conclusion
transesterification process improved the fuel properties of the oil
with respect to specific gravity, density, viscosity, pour point, This research revealed that, biodiesel can be produced by
and cloud point. The comparison of these properties with in situ (direct) transesterification method from the seeds of
diesel shows that, the methyl ester has a relatively closer fuel rubber tree, and it was found that, the maximum biodiesel yield
property values to that of diesel. Viscosity is the most obtained was 96 % at 60 °C with mass ratio of 1:6, 3.0 g of KOH
important property of biofuel since it affects the operation of fuel as catalyst, stirring speed of 500 rpm for 120 min. The results
injection equipment, particularly at low temperatures where the were also in agreement with the findings of a previous study by
increase in viscosity affects the fluidity of the fuel 25). Biodiesel Widayat et al. 3) on Production Process of Biodiesel from Rubber
has a viscosity close to that of diesel fuels. High viscosity leads Seeds (Hevea
to poorer

Table 2 Properties of methyl esters of rubber seed in comparison with other

esters
Biodiesel
Testing standard Rape seed Soybean oil RSO RSO RSO
S/N Property procedure This work oil FAME 4) FAME 4) FAME 14) FAME 4) FAME 24)
ASTM 6751-02

1 Specific gravity at 20 °C ASTM D4052 0.87–0.90 0.874 0.882 0.885 - 0.874 0.881
2 Density at 20 °C (g/cm3) ASTM D4052 - 0.897 NA NA - - -
3 Viscosity (mPa s) at 40 °C ASTM D445 1.9–6.0 4.68 4.5 4.08 3.89 5.81 4.059
4 Pour point (°C) ASTM D97 -15 to 10 -4.0 -12 -3 -2.0 -8.0 -
5 Cloud point (°C) ASTM D2500 -3 to 12 4.8 -4.0 -2.0 3.2 4.0 -
6 Acid value (mg KOH/g) ASTM D974 0.8 0.65 NA NA 0.42 0.11 0.22
 768 J. Jpn. Inst. Energy, Vol. 94, No. 7, 2015

Brasiliensis) by In Situ Transesterification Method with

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