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Kinetic studies and thermodynamics of oil extraction

and transesterification of Chlorella sp. for biodiesel
production
a a a a
A.L. Ahmad , N.H. Mat Yasin , C.J.C. Derek & J.K. Lim
a
School of Chemical Engineering, Engineering Campus, Universiti Sains Malaysia, Seri
Ampangan, 14300 Nibong Tebal, Seberang Perai Selatan, Pulau Pinang, Malaysia
Published online: 19 Nov 2013.

To cite this article: A.L. Ahmad, N.H. Mat Yasin, C.J.C. Derek & J.K. Lim (2014) Kinetic studies and thermodynamics of oil
extraction and transesterification of Chlorella sp. for biodiesel production, Environmental Technology, 35:7, 891-897, DOI:
10.1080/09593330.2013.855263

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 Environmental Technology, 2014
Vol. 35, No. 7, 891–897, http://dx.doi.org/10.1080/09593330.2013.855263

Kinetic studies and thermodynamics of oil extraction and transesterification of Chlorella sp.
for biodiesel production
A.L. Ahmad∗ , N.H. Mat Yasin, C.J.C. Derek and J.K. Lim
School of Chemical Engineering, Engineering Campus, Universiti Sains Malaysia, Seri Ampangan, 14300 Nibong Tebal,
Seberang Perai Selatan, Pulau Pinang, Malaysia
(Received 26 June 2013; final version received 8 October 2013 )

In this work, a mixture of chloroform and methanol (1:1, v/v) was applied to oil extraction from Chlorella sp. at 30,
40, 50 and 60◦ C for 150 min extraction times. Kinetic studies revealed that the values of n and the rate constants were
Downloaded by [The University of Manchester Library] at 07:06 19 October 2014

found to depend strongly on temperature. The activation energy was Ea = 38.893 kJ/mol, and the activation thermodynamic
parameters at 60◦ C were S = = −180.190 J/mol K, H = = 36.124 kJ/mol and G = = 96.128 kJ/mol. Both H and
S yielded positive values, whereas G was negative at 60◦ C, indicating that this process is endothermic, irreversible and
spontaneous. The acidic transesterification process was also investigated by gas chromatographic analysis of the microalgae
fatty acid methyl esters (biodiesel) at different temperatures and reaction times. The fatty acid profile indicated that the
main components were palmitic, linoleic and linolenic acids. The concentration of linolenic acid increased and oleic acid
decreased as the temperature increased. Two-hour transesterification is the best reaction time for biodiesel production because
it produces the highest percentage of unsaturated fatty acids (74%). These results indicate the potential of Chlorella sp. to
produce biodiesel of good quality.
Keywords: microalgal biomass; kinetics; thermodynamics; oil extraction; biodiesel

1. Introduction more interest in the cultivation of microalgae because of the

Biodiesel has been receiving considerable attention in
recent years due to its potential as a sustainable, biodegrad-
able, nontoxic fuel and environmental-friendly alterna-
tive to petrodiesel.[1] Biodiesel derived from oil crops
is potentially renewable and a carbon-neutral alterna-
tive to petroleum fuels. Unfortunately, biodiesel from oil
crops, waste cooking oil and animal fat cannot realisti-
cally satisfy even a small fraction of the existing demand
for transportation fuels. The relatively high cost of this
biodiesel renders the resulting fuels unable to compete
with petroleum-derived fuel. Raw materials and ways to
reduce the production costs of biodiesel should, therefore,
be explored.
Microalgae (microscopic unicellular organisms with
oxygenic photosynthetic capabilities) appear to be the
only source of renewable biodiesel that has the poten-
tial to replace the petroleum-derived transportation fuels
completely. Microalgae are a specialized group of microor-
ganisms that live in diverse ecological habitats such as
freshwater, brackish and marine ecological systems. Similar
to plants, microalgae use sunlight to produce oils, but they
do so more efficiently than crop plants. Oil productivity
from many microalgae greatly exceeds the oil productiv-
ity of the best-producing oil crops. Researchers are gaining
valuable metabolites and lipids that can be found in them.
The advantages of culturing microalgae as a resource for
biomass are listed elsewhere.[2–6]
Various methods to extract the oil from algae, such
as expeller/press, solvent extraction and supercritical fluid
extraction, are available.[7] Extraction using supercritical
fluid provides very high purity and good product concentra-
tion. However, the operating and investment cost is high.[8]
Solvent extraction, a common and efficient technique for
producing oil for biodiesel production, involves the trans-
fer of a soluble fraction from a solid material to a liquid
solvent. Solvent extraction has a relatively low operating
cost compared with supercritical fluid extraction. There are
three methods that have been identified for solvent extrac-
tion: (i) hot water extraction, (ii) Soxhlet extraction and (iii)
ultrasonication technique.[9]
However, the solvent extraction has some disadvantages
such as poor extraction of polar lipids, long time required
for extraction and hazards of boiling solvents.[10] In fact,
the lipid extraction step from microalgae still represents a
bottleneck of the whole process of producing biodiesel from
microalgae. In particular, the main limitation related to the
use of organic solvents for extracting lipids from microal-
gae is their high cost and the need of their continuous supply

∗ Corresponding author. Email: chlatif@eng.usm.my

© 2013 Taylor & Francis

 892 A.L. Ahmad et al.
since not all of them can be suitably recycled. For this rea-
son, a rigorous quantification of the extraction kinetics is
critical to optimize the extraction process and consequently
increase the economical feasibility of the process. More-
over, the organic solvents used to perform extraction are
typically toxic and thus the optimization of the amounts
used during the extraction step might result in a higher envi-
ronmental sustainability of the process. The most popular
chemical for solvent extraction is hexane, which is rela-
tively inexpensive. Suganya and Renganathan [11] studied
the efficiency of algal oil extraction using six different
extraction methods with 12 different solvent systems and
showed that the extraction of oil using an ultrasound pre-
treatment method with 1% diethyl ether and 10% methylene
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chloride in hexane achieved a high yield of oil.
Numerous methods are currently available and have
been adopted for the production of biodiesel fuel. The four
primary ways and processes to produce biodiesel include
direct use and blending, microemulsion, thermal crack-
ing (pyrolysis) and transesterification.[12,13] The most
common method for converting the oils to biodiesel is trans-
esterification due to its low cost and simplicity. Methanol
is usually the preferred alcohol for producing biodiesel
because of its low cost and its physical and chemical
advantages.
In the present investigation, oil extraction from
Chlorella sp. with hexane with acidic transesterification for
converting the oils to biodiesel was studied. The objectives
of this study were to (i) determine the rate constant and
activation energy; (ii) determine the enthalpy value and
other thermodynamic parameters for oil extraction from
Chlorella sp. biomass and (iii) study the effect of temper-
ature and reaction time for acidic transesterification on the
fatty acid (biodiesel) yield.
2. Materials and methods
2.1. Preparation of microalgae biomass
Chlorella sp. belongs to the family of green algae and is
considered part of phytoplankton.[14] Chlorella sp. is the
strain most favoured by researchers for biodiesel produc-
tion and has been described as a good option for biodiesel
production.[15] Chlorella sp. was grown in Bold’s Basal
Medium and incubated in 2 l Erlenmeyer flasks filled with
2000 ml distilled water for 14 days of cultivation. The
medium and flasks were sterilized in an autoclave for
15 min at 121◦ C to prevent any contamination during the
early stages of culture growth. The details of the specific
measurements and characterization of the Chlorella sp.
cultures were reported in our previous research.[15–17]
After growth, Chlorella sp. was harvested by centrifuga-
tion at 4000 rpm for 10 min using a model Megafuge 40
centrifuge (Thermo Scientific, Germany) and then dried
overnight with a freeze dryer (model 7754030, Labconco,
US).
2.2. Microalgal cells wall disruption
Freeze-dried samples of 20 mg of Chlorella sp. biomass
along with 5.5 ml distilled water were placed into capped
test tubes and then sonicated (model 8510, Branson, USA)
for 5 min. The optical microscope (model BX53F, Olympus,
Japan) was used to characterize the Chlorella sp. biomass
before and after sonication to identify the changes in the
shape of the Chlorella sp. cells.
2.3. Lipid extraction
The total lipids were extracted by a mixture of
chloroform/methanol (1:1, v/v) using a slightly modified
version of Bligh and Dyer’s method.[18] Approximately
12 ml of the mixed solvents was used in each extraction
step. The extraction was executed on a water bath preheated
to the set temperatures (30, 40, 50 and 60◦ C) on a heating
plate using a magnetic stirrer. The total extraction time was
150 min with 30 min time intervals. All experiments were
conducted in triplicate.
2.4. Determination of oil extraction yield
The amount of oil extracted relative to the time at different
temperatures was determined gravimetrically by measuring
the weight of the residue after drying the solvent under a
flow of nitrogen gas. The oil extraction yield (% w/w) was
calculated using the following equation:
Weight of oil extracted (g)
Oil extraction yield (%) = × 100
Weight of biomass (g)
(1)
2.5. Transesterification
In this experiment, temperature and reaction time were
the important factors in the transesterification process. The
molar ratio of methanol to oil (56:1) and the mixing rate
were fixed. An acid catalyst (hydrochloric acid) was used
in this research because the alkali catalyst was not suit-
able for the transesterification of microalgal oil, most likely
because of the high acid value of microalgal oil.[19] Zhang
et al.[20] also reported that the alkali catalytic process is
sensitive to water. The presence of water under alkaline con-
ditions may cause ester saponification. A standard reaction
mixture consisting of 4.25 ml methanol, 5 ml hexane and
215 μl HCl (37% vol.) was added to the oil and mixed at
500 rpm. After heating for the specified period, the reaction
mixture was allowed to cool and then centrifuged at 3000
rpm for 2 min, resulting in the separation of the two layers.
The upper layer consisted of methyl esters (biodiesel), and
the lower layer contained the glycerol, the excess methanol
and the remaining catalyst. Each experiment was conducted
in triplicate. The biodiesel from each experiment was anal-
ysed individually in triplicate to calculate the mean values
of the experimental data.
 Environmental Technology
2.6. Gas chromatographic analysis
A fatty acid composition analysis was performed using a gas
chromatograph (model 7890A, Agilent, China) equipped
with a flame ionization detector, splitless automatic injec-
tor and fused silica capillary column (30 m × 0.25 mm ×
0.25 μm) (Omegawax 250, Supelco, USA). Injector and
detector temperatures were kept constant at 250 and 280◦ C,
respectively. The oven temperature program started at
100◦ C for 1 min and then ramped to 190◦ C at 25◦ C/min and
remained constant at this temperature for 20 min. Each fatty
acid component was identified by comparing the retention
time and peak area with the retention time and peak area in
the standard solutions, and the composition was calculated
as the percentage of the total fatty acids present in the sam-
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ple. Six fatty acids (C16:0, C16:1, C18:0, C18:1, C18:2 and
C18:3) were used as the standard materials. Three replicates
of each fatty acid analysis were performed.
3. Results and discussion
3.1. Sonication pretreatment
To increase the cell wall damage in the Chlorella sp. cell,
a sonication pretreatment method was implemented on the
Chlorella sp. biomass before the oil extraction. The advan-
tages of this sonication method are reduced extraction time,
reduced solvent consumption, greater penetration of solvent
into cellular materials and improved release of cell contents
into the bulk medium.[9] The method also disrupts the cell
wall, leading to an increase in the oil extraction yield from
the Chlorella sp. biomass due to the penetration of solvent
(a) (b)
Figure 1. Optical observation of Chlorella sp. cells (a) before
and (b) after sonication (Mag = 20×).
Table 1. The oil extraction yield (%) from Chlorella sp. biomass and the reaction rate constants at different temperatures with respect
to extraction time.
Temp Reaction rate constant,
(◦ C) k (min−1 ) n 30 min
30 4.7 × 10−4 1.638 8.43 ± 0.14
40 1.12 × 10−3 1.488 9.65 ± 0.11
50 1.23 × 10−3 1.504 10.48 ± 0.05
60 2.1 × 10−3 1.363 10.89 ± 0.07
Note: Values are means ± standard deviations of three replicates.
893
into the inner surfaces of the Chlorella sp. cells. In addi-
tion, the Chlorella sp. cells were also dispersed into smaller
structures after sonication method as shown in Figure 1.
3.2. Extraction kinetics
A reaction rate equation for oil extraction from Chlorella
sp. biomass can be written as [21]
dY
= kY n (2)
dt
where Y is the oil extraction yield (%), t is the time of
extraction (min), k is the extraction rate constant (min−1 )
and n is the order of the reaction. As the percentage of oil
extraction increases over time, the term dY /dt has a positive
sign.[21]
Using the value in Table 1 and applying the differential
method, plots of ln (dY /dt) versus ln Y at different temper-
atures were found to be linear according to Equation (2).
The order of the reaction was determined from the slopes
of the straight lines in Figure 2, and the reaction rate con-
stants were calculated from the intercept of the linear plot.
A fractional kinetic (with order of about 1.5) was found
from the values of n obtained with an average regression
coefficient R2 = 0.969. The obtained values of n vary from
a minimum of 1.363 to a maximum of 1.638 as shown in
Figure 2 and Table 1. From the analysis of the data, the oil
extraction yield was found to increase with an increase in
extraction time and temperature, showing the influence of
these two factors on lipid extraction. The reaction rate con-
stant, k, the time required to obtain maximum extraction of
oil from the Chlorella sp. biomass, increases with increas-
ing temperature, and this trend is obvious in Table 1. This
observation appears to suggest that the reaction rate con-
stants are often found to depend strongly on temperature.
This trend was also found by Suganya and Renganathan
with marine macroalgae strain.[11] These authors reported
that an increase in the reactivity of the solvent enhances the
rate of extraction. However, in their works, the increase in
k values is significant at the temperature higher than 45◦ C.
As can be noticed, in comparison with other strains such as
sunflower seeds [21] and Jatropha Curcas,[22] an increase
in k values is consistent even at high temperature. This phe-
nomenon is similar with our findings; there were very slight
Oil extraction yield (%)
60 min 90 min 120 min 150 min
8.93 ± 0.10 9.49 ± 0.05 10.11 ± 0.012 10.79 ± 0.14
10.73 ± 0.30 12.24 ± 0.14 13.96 ± 0.15 15.96 ± 0.15
12.14 ± 0.20 13.98 ± 0.19 16.37 ± 0.10 19.78 ± 0.10
12.81 ± 0.04 15.64 ± 0.22 19.54 ± 0.18 24.04 ± 0.37
 894 A.L. Ahmad et al.

Table 2. The thermodynamic activation parameters for

Chlorella sp. biomass oil extraction at various temperatures.

Temp
(K) S = (J/mol K) H = (kJ/mol) G = (kJ/mol)

303 −179.405 36.374 90.733

313 −179.675 36.291 92.529
323 −179.936 36.208 94.327
333 −180.190 36.124 96.128

theory [21]:
Figure 2. A plot of ln (dY /dt) versus ln Y at different tempera- RT S = /R
tures for oil extraction from Chlorella sp. biomass. The error bars A= e (4)
indicate the standard deviations among the replicates. Nh
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H = = Ea − RT (5)
G = = H = − T S = (6)

where N is Avogadro’s constant, h is Planck’s constant,

S = is the activation entropy (J/mol K), H = is the
activation enthalpy (kJ/mol) and G = is the activation
free energy or Gibb’s energy (kJ/mol). These activation
thermodynamic parameters are shown in Table 2 for each
temperature.

3.5. Calculation of thermodynamic parameters

Figure 3. Activation energy calculation from the plot of ln k
versus 1/T (K−1 ).
difference in the results of k values at the temperature of 40,
50 and 60◦ C as shown in Table 1.
3.3. Calculation of activation energy
The change in the rate constant can be described by the
Arrhenius equation [21]:
k = Ae−Ea/RT
where k is the reaction rate constant (min−1 ), A is the Arrhe-
nius constant (s−1 ), Ea is the activation energy (kJ/mol), R
is the universal gas constant (J/mol K) and T is the absolute
temperature (K). A plot of ln k versus 1/T yields a straight
line with the slope representing the activation energy of
extraction (-Ea/R) and an intercept as the Arrhenius con-
stant, ln A (Figure 3). The activation energy, Ea, and the
Arrhenius constant, A, were calculated as 38.893 kJ/mol
and 2684 s−1 , respectively.
3.4. Calculation of thermodynamic activation
parameters
The thermodynamic activation parameters (S = , H =
and G = ) for microalgal oil extraction were calculated
using the following equations according to transition state
The thermodynamic parameters (S, H and G) for the
extraction of microalgal oil can be estimated using the
expressions below [21]:
YT
K= (7)
Yu
G 1 H 1 S
ln K = − =− + (8)
R T R T R
where K is the equilibrium constant, YT is the percent oil
yield (%) at t = 150 min at temperature T , Yu is the percent
unextracted oil (%), H is the enthalpy change (kJ/mol),
S is the entropy change (1/mol K) and G is the free
(3)
energy or Gibb’s energy (kJ/mol). At equilibrium, none of
the oil yields changes with time.
The plot of ln YT versus 1/T at 150 min yields a straight
line of ln YT = −HR(1/T ) + C with the slope repre-
senting the enthalpy change of extraction, – HR. The
enthalpy change was calculated to be H = 0.308 kJ/mol
for Chlorella sp. biomass oil extraction (Figure 4). The H
value obtained indicated the physicochemical nature of the
oil extraction process.[21] The positive value for enthalpy
indicates that the oil extraction process is endothermic and
requires energy input during the process.
Other thermodynamic parameters (S and G) and
the equilibrium constant values for Chlorella sp. biomass
oil extraction are given in Table 3 for each temperature.
An increase in the entropy change and a decrease in the
free energy indicate that the process is spontaneously for-
ward. The positive value of entropy change (S > 0) at
 Environmental Technology 895

summarized in Table 4. Based on the previous report,[24]

the most common fatty acids contained in biodiesel were
palmitic, stearic, oleic and linolenic acid. However, in our
study on the Chlorella sp. biomass, palmitic acid (C16:0),
linoleic acid (C18:2) and linolenic acid (C18:3) were the
major components, followed by stearic acid (C18:0), oleic
acid (C18:1) and small amounts of palmitoleic acid (C16:1)
was also identified. To study the effect of temperatures,
experiments were carried out at temperatures of 70, 80
and 90◦ C because the transesterification is not affected at
low temperature. This is in agreement with the results pre-
sented in the previous research by Rashid and Anwar [25];
Figure 4. Enthalpy change calculation from the plot of ln YT
an increase in the reaction temperature leads to a higher
versus 1/T (K−1 ).
yield of fatty acid. In spite of this, good quality of biodiesel
Downloaded by [The University of Manchester Library] at 07:06 19 October 2014

Table 3. The equilibrium constant (K) and the thermody- could be obtained at a higher temperature. However, at very
namic parameters (S and G) for Chlorella sp. biomass high temperature (above 90◦ C) it could burn some of the
oil extraction at different temperatures. oil before completion of the alcoholysis. Thus, only three
temperatures were chosen in this experiment. Unlike the
Temp
(K) K G (kJ/mol) S (1/mol K) two major fatty acids, special attention should be taken to
the content of linolenic acid (C18:3) because the European
303 0.372 2.494 −7.214 regulation EN 14103 has set 12% as the maximum allowed
313 0.669 1.048 −2.362 content of C18:3 for quality biodiesel. Furthermore, fatty
323 0.987 0.036 0.841
333 1.522 −1.164 4.420 acid with a high oleic acid (C18:1) has been reported to have
a reasonable balance of fuel properties due to increases of
oxidative stability for longer storage.[26,27] As shown in
T ≥ 50◦ C indicates that the process is irreversible, whereas Table 4 linolenic acid content increased as reaction tem-
the value of S is negative for the T < 40◦ C. Thus, the perature increased, while opposite trend was observed in
extraction process at the lower temperature is not suitable oleic acid content. These findings differ from the reported
for oil extraction of Chlorella sp. because the equilibrium lipid content of Nannochloropsis oculata and Chlorella vul-
constant is very low at the temperature lower than 40◦ C as garis, where the increase in temperature led to an increased
shown in Table 3. The different types of results were previ- content of oleic acid (C18:1).[14] Therefore, these two fatty
ously reported by Amin et al.[22] and Liauw et al.[23] for acids (C18:1 and C18:3) might be useful as a good indicator
the extraction of oil from Jatropha Curcas and Azadirachta for various temperatures.
indica A. Juss. In their works, S is positive and G is The influence of reaction time was also analysed by
negative for all conditions. The equilibrium constant is also varying the reaction time up to 5 hours while keeping the
high (K > 1) even at low temperature. optimized temperature at 90◦ C. The trends observed when
varying the reaction time from 1 to 5 hours were gener-
3.6. Fatty acid compositions ally inconsistent for most fatty acids. As noted by Knothe
The fatty acid composition of the Chlorella sp. was tested at [27] and Griffiths and Harrison,[28] the carbon chain length
various temperatures and reaction times, and the results are and the degree of unsaturation are important characteristics

Table 4. Summary of fatty acid compositions in Chlorella sp. at various temperatures and reaction times (unit % of total fatty acid).

Temperature (◦ C) Reaction time (h)

Fatty acids 70 80 90 1 2 3 4 5

C16:0 29.07 ± 0.51 30.08 ± 1.89 26.01 ± 0.12 28.41 ± 4.11 24.25 ± 1.56 37.56 ± 2.90 25.15 ± 1.37 28.41 ± 3.58
C16:1 0.83 ± 0.04 0.89 ± 0.09 2.62 ± 0.08 0.55 ± 0.33 0.75 ± 0.28 1.09 ± 0.17 1.41 ± 0.34 1.73 ± 0.57
C18:0 9.48 ± 0.57 12.77 ± 0.17 3.01 ± 0.28 1.82 ± 0.19 1.50 ± 0.00 1.58 ± 0.44 1.78 ± 0.45 2.21 ± 0.23
C18:1 2.66 ± 0.22 2.55 ± 0.14 2.44 ± 0.05 3.04 ± 0.02 4.73 ± 1.17 2.87 ± 0.36 4.12 ± 1.10 4.14 ± 0.85
C18:2 38.75 ± 0.96 33.49 ± 0.74 39.45 ± 1.46 42.53 ± 0.58 45.59 ± 3.58 36.83 ± 4.30 42.26 ± 2.41 41.01 ± 4.57
C18:3 19.22 ± 0.64 20.22 ± 3.78 26.49 ± 1.19 23.65 ± 2.61 23.17 ± 0.28 20.07 ± 0.07 25.28 ± 1.76 22.51 ± 1.76
SFA (%) 38.55 ± 0.29 42.85 ± 1.12 29.01 ± 0.11 30.22 ± 2.53 25.75 ± 1.35 39.14 ± 1.28 26.93 ± 1.31 30.62 ± 1.58
UFA (%) 61.45 ± 0.59 57.15 ± 1.43 70.99 ± 1.31 69.78 ± 0.26 74.25 ± 3.1 60.86 ± 1.51 73.07 ± 1.28 69.38 ± 2.35

SFA, saturated fatty acids; UFA, unsaturated fatty acids.

Note: Data are expressed as mean ± standard deviations of three replicates.
 896 A.L. Ahmad et al.
of fatty acids for biodiesel production and may affect the
quality of the biodiesel produced. Table 4 shows that the
Chlorella sp. lipids for all reaction times are composed
mainly of unsaturated fatty acids (60–75%), with a signif-
icant percentage of palmitic acid also present (25–38%).
Among the five reaction times applied in our study, the
shortest reaction time (2 h) shows the highest percentage of
UFAs, and special attention should be paid to the linoleic
acid because the C18:2 unsaturated chains were the pre-
dominant fatty acids (45.6%) followed by C18:3 (25.2%)
and C16:0 (24.3%). It is also important to highlight that
higher compositions of UFAs can reduce the pour point
of biodiesel and making it feasible to be used in cold
climate countries.[29] However, when the UFA with dou-
Downloaded by [The University of Manchester Library] at 07:06 19 October 2014
ble bond ≥ 3, their high content is undesirable since it
reduces the chemical and oxidative stability of the resulting
biodiesel.[27] Therefore, the higher content of unsaturated
chains (such as C18:1 and C18:2) and the lower content
of C18:3 in Chlorella sp. at 2 h of reaction time may be
expected to produce a good quality of biodiesel. Further
study will be necessary to uncover the feasibility and effec-
tiveness of various harvesting methods for the production
of high-quality biodiesel from Chlorella sp., then the phys-
ical and fuel properties may be comparable to diesel fuel
and the ASTM biodiesel standard.
Conclusion
The kinetics for oil extraction from Chlorella sp. biomass
by a mixture of chloroform/methanol (1:1, v/v) were found
to be a fractional kinetic with the order of about 1.5 by
the differential method and strongly dependent on tem-
perature. The activation energy was Ea = 38.893 kJ/mol
and the thermodynamic activation parameters at 60◦ C
were S = = −180.190 J/mol K, H = = 36.124 kJ/mol
and G = = 96.128 kJ /mol. The enthalpy value was found
to be positive (H = 0.308 kJ/mol), indicating that the
process is endothermic and requires the input of energy
during the process. The other thermodynamic parameters
at 60◦ C were calculated to be S = 4.420 1/mol K and
G = −1.164 kJ/mol. Our study also demonstrated that
palmitic acid (C16:0), linoleic acid (C18:2) and linolenic
acid (C18:3) are major fatty acids found in Chlorella sp.,
indicating a good potential for use of the Chlorella sp. lipids
in the production of biodiesel. The variation of temperature
influenced the oleic acid (C18:1) and linolenic acid (C18:3)
contents strongly, as an increase from 70 to 90◦ C increased
the content of linolenic acid from 19.22 to 26.49% and
decreased the oleic acid from 2.66 to 2.44%. Finally, 2 h
of reaction time yields the most appropriate fatty acid com-
position in terms of unsaturated fatty acids such as C18:1
and C18:2. The next research effort will address the har-
vesting of Chlorella sp. cells using appropriate and efficient
methods, making Chlorella sp. biomass a good candidate
for biodiesel production.
Acknowledgements
All authors are affiliated with the Membrane Science and Tech-
nology Cluster USM.
Funding
This material is based on the work supported by a Research
University (RU) [Grant No. 1001/PJKIMIA/814060) and a Post-
graduate Research (PRGS) [Grant No. 1001/JKIMIA/8044014]
from Universiti Sains Malaysia (USM). N.H. Mat Yasin grate-
fully acknowledges Universiti Malaysia Pahang [SLAB 2011] for
a scholarship.
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