Sustainable Environment Research 28 (2018) 444e453

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Original Research Article

Optimization and characterization of bio-oil produced from Ricinus

communis seeds via ultrasonic-assisted solvent extraction through
response surface methodology
Chiena L. Palconite a, Alexandra C. Edrolin a, Sheryl Nope B. Lustre a, Aldin A. Manto a,
John Rey L. Caballero a, Maribel S. Tizo b, Alexander L. Ido a, Renato O. Arazo a, c, *
a
College of Engineering and Technology, University of Science and Technology of Southern Philippines, Claveria 9004, Philippines
b
College of Arts and Sciences, University of Science and Technology of Southern Philippines, Claveria 9004, Philippines
c
Project Development Office, University of Science and Technology of Southern Philippines, Claveria 9004, Philippines

a r t i c l e i n f o a b s t r a c t

Article history: As the world confronts a surging price of fuel along with various environmental issues, researchers
Received 20 March 2018 around the globe are scrambling to identify and develop potential biofuel feedstocks that fall outside the
Received in revised form “food or fuel dilemma”. This justifies the need for non-edible oilseeds that can be used as sustainable
30 June 2018
feedstock for biofuel production. In this study, non-edible Ricinus communis seeds were utilized in the
Accepted 27 July 2018
Available online 6 October 2018
production of bio-based fuel to support the crusade for the use of renewable energy. Extraction of bio-oil
from R. communis seeds was done through ultrasonic-assisted solvent extraction. The effects of n-hexane
to biomass (HB) ratio, extraction time, and resonance amplitude on bio-oil yield were investigated.
Keywords:
Ricinus communis
Optimization of bio-oil yield was done through central composite design of the response surface
Bio-oil methodology. The optimum bio-oil yield was found to be 42% at optimum conditions of 10:1 HB ratio,
Optimization 25 min extraction time and 50 mm resonance amplitude. Characterization of the extracted bio-oil
Ultrasonic-assisted solvent extraction revealed a density of 0.9 g mL
1, kinematic viscosity of 18.6 cSt at 40  C, and higher heating value of
Response surface methodology 29.5 MJ kg
1. The Fourier Transform Infrared Radiation spectroscopy analysis of the bio-oil revealed that
it contained ester functional groups. Gas chromatography analysis showed that the major fatty acids
present in the bio-oil are palmitic (2.2%), stearic (2.4%), oleic (6.7%), linoleic (8.8%) and ricinoleic (77.1%).
The R. communis bio-oil is enriched with unsaturated fatty acids while there is a relatively low content of
saturated fatty acid accounting 87.8% and 11.3% of the total fatty acids composition, respectively. As
confirmed by this study, the results indicate the potential of R. communis bio-oil as a source for biodiesel
production.
© 2018 Chinese Institute of Environmental Engineering, Taiwan. Production and hosting by Elsevier B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).

1. Introduction However, the extensive consumption of fossil fuels brought twin

Energy has become an essential commodity for humanity to
continue economic growth and maintain a high standard of living.
Fossil fuels provide most of this energy that powers modern soci-
ety. The increasing high energy demand in the industry and
transportation sectors led to the widespread use of fossil fuels.
* Corresponding author. College of Engineering and Technology, University of
Science and Technology of Southern Philippines, Claveria 9004, Philippines.
E-mail address: roarazo@yahoo.com (R.O. Arazo).
Peer review under responsibility of Chinese Institute of Environmental
Engineering.
crises of source depletion and environmental degradation [1].
These encourage for the development of renewable energy sources
which can supplement or replace fossil fuels.
A promising alternative energy source is the utilization of
biomass to produce bio-oil, a potential feedstock for biodiesel
production for three main reasons [2]. First, it is a renewable energy
source that could be sustainably developed in the future. Second, it
could contribute to a healthy environment since it reduces green-
house gas emissions by displacing the use of fossil fuels. Third, it
appears to have significant economic potential provided that fossil
fuel prices will continue to increase in the future. Currently,
increasing percentage is mandated in many parts of the world to

https://doi.org/10.1016/j.serj.2018.07.006
2468-2039/© 2018 Chinese Institute of Environmental Engineering, Taiwan. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND
license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453
blend diesel fuel with biodiesel. In the Philippines, a minimum of
5% blend of biodiesel in all diesel products was mandated starting
the year 2015 through Biofuels Act of 2006 [3] and directed go up to
20% in the year 2030 [4].
However, bio-oils have been strongly criticized recently since
these are mainly produced from edible sources or first-generation
feedstocks. These raise many concerns such as the continuing
food versus fuel debate [5]. Also, it has been found that feedstock
alone accounts approximately 75% of the overall production cost
[6]. Thus, it is crucial to employ inexpensive feedstocks to replace
expensive refined oils.
Production of bio-oil from non-edible sources or second-
generation feedstocks can overcome the problems of food versus
fuel dispute and economic issues related to edible sources. Recently,
these bio-oils from non-edible sources have gained enormous
attention because of high bio-oil content, easy availability, and
having the advantage that it could be grown on lands which are not
suitable for agriculture. Additionally, it could be cultivated with less
intensive attention; thus, reducing the cost of production [6]. Hence,
many researchers have concluded that the use of non-edible feed-
stocks for biodiesel is sustainable and viable alternative [7,8].
The seeds of Ricinus communis (commonly known as castor) is
considered as a second-generation feedstock. It has limited applica-
tions to human and animal considering that it contains ricin, a highly
water soluble toxin [9]. However, it has comparative advantages to
other seeds when used as feedstock for oil production. The plantation
cost of R. communis is significantly less than other plants such as
jatropha, soybean, and rapeseed. R. communis plant is also drought
tolerant, pest-resistant, require less artificial care, and biomass could
be harvested three times in a year. Therefore, R. communis seed is a
sustainable source and an attractive feedstock for bio-oil production.
There are already published studies on biodiesel production
using R. communis seeds [10e12]. However, there is no current
report on the extraction of bio-oil from R. communis seeds through
ultrasonic-assisted solvent extraction (UASE) to maximize its po-
tential as possible liquid feedstock for biodiesel production. The
primary aim of this study was to optimize the extraction of
R. communis seed bio-oil through UASE using response surface
methodology. UASE was applied since this technique was found to
reduce the extraction time and solvent consumption but at the
same time possessing higher extraction efficiency than the con-
ventional methods [13]. Needless to say, this study aimed to
effectively make use of the underutilized R. communis seeds as
feedstock for the production of bio-oil and the subsequent pro-
duction of refined biodiesel.
2. Materials and methods
2.1. Chemicals
The n-hexane (95%, GR grade, Duksan Pure Chemicals) used in
this study was purchased from Harnwell Chemical Corporation,
Cagayan de Oro City, Philippines.
2.2. Biomass preparation and characterization
The seeds were obtained from R. communis plants growing wild
on marginal lands around Jasaan, Misamis Oriental, Philippines.
Harvested R. communis fruits were manually cleaned and sun-dried
for easy discharge of the encased seeds. The seeds were separated
from the pods and dried at 80  C for 7 h in a hot air oven. The mass
of the seeds was recorded for the “as received” moisture content
analysis. Then, the seeds' shells were cracked, and the kernels were
ground to a paste using mortar and pestle and oven dried further at
100  C for 4 h before extraction. The final mass of the seeds was
445
recorded again for the moisture content (dry basis) analysis. In ash
content determination, the temperature was set at 600  C and
allowed to stay for 3 h inside a muffle furnace. Moisture and ash
contents were analyzed using the oven (Contherm Designer Series
Oven, Model: CE Z011) and muffle furnace (LabTech LEF-304P-2),
respectively. Results were presented as percentage relative to the
mass of the R communis seeds sample used.
2.3. Experimental design for ultrasonic-assisted solvent extraction
Design Expert 7.0 software was used to design the experiments
and analyze the results of bio-oil extraction stage. Central com-
posite design (CCD) of the response surface methodology (RSM)
was applied to the experiment. In the design of experiments, three
important factors such as n-hexane to biomass (HB) ratio, extrac-
tion time and resonance amplitude were selected. In the pre-
liminary study, experiments were carried out to extract bio-oil from
R. communis seeds (15 g) using n-hexane as solvent for extraction
times of 20, 25, 30, 35 and 40 min with HB ratios of 7:1, 8:1, 9:1, 10:1
and 11:1, and various resonance amplitudes of 40, 45, 50, 55 and
60 mm. The HB ratio, extraction time and resonance amplitude were
chosen based on recent literature [12,14,15]. The succeeding
experimental runs were based on the parametric study principle of
taking the center of one variable as constant and combining it to all
ranges of the other variables and finally determined a peak in a
once plotted graph. This stage was done to explore the wide range
of reaction conditions and identify the most probable value of the
operating variables to be considered in optimization study.
2.4. UASE setup and experimental procedure
R. communis seed paste was put in an improvised jar containing
n-hexane which was then placed in the fume hood (Fig. 1). The
extraction process was done by inserting directly the ultrasonic
probe into the mixture according to the predetermined HB ratio,
ultrasonication time and resonance amplitude. The samples were
extracted under continuous ultrasonic waves. This study used an
ultrasonic processor UP400S (400 W, 24 kHz). The processor is
45 mm long and has 22 mm diameter tip sonotrode with
85 W cm
2 acoustic power density, and a maximum amplitude of
100 mm. During the extraction, the temperature was maintained at
60 ± 5  C.
After each run, the reactor was cooled down. After which, 60 mL
distilled water was added to the mixture and was mixed slowly
with a stirring rod. Within 3 h, the mixture was set aside to form a
biphasic layer. This upper phase is composed of n-hexane and
R. communis seed bio-oil, respectively. The upper phase was care-
fully decanted, and it was transferred to a beaker. Then, a 20 mL n-
hexane was added to the cell residue and stirred to extract recov-
erable bio-oil. For 10 min, the mixture was set aside. Then, the n-
hexane and R. communis seed bio-oil were decanted and added to
n-hexane - R. communis seed bio-oil suspension, then heated. The
heating temperature was at 70 ± 5  C letting n-hexane be evapo-
rated while the bio-oil remained.
2.5. Bio-oil yield determination
The yield of R. communis seed bio-oil was calculated using Eq.
(1) and expressed as a percentage of the initial mass of biomass:
mass of bio
oil extractedðgÞ
% Bio
oil yield ¼ x 100 (1)
mass of initial biomassðgÞ
446 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453
Fig. 1. Experimental setup for the UASE.
2.6. Analytical methods
The bio-oil extracted from R. communis seeds had undergone
analysis by different tests to determine if the product met the
required specifications. The density of the extracted bio-oil was
computed using the mass and volume of the sample. Kinematic
viscosity was analyzed following ASTM D-445-71 (Kinematic Vis-
cosity of Transparent and Opaque Liquids) using Glass Capillary
Viscometer. The high heating value was determined using ASTM
D4809 (Standard Test Method for Heat of Combustion of Liquid
Hydrocarbon Fuels by Bomb Calorimeter Precision Method). The
functional groups were determined through Fourier Transform
Infrared Radiation (FTIR) spectroscopy analysis. The fatty acid
profiling was done through gas chromatographyemass spectros-
copy (GCeMS) analysis following Official Method of Analysis of
AOAC International, 19th ed., 2012.
3. Results and discussion
3.1. Characteristics of R. communis seeds
Table 1 shows the initial characteristics of R. communis seeds.
The result of the moisture content analysis of 1.9 ± 0.0% (dry basis)
agreed well with the literature where moisture contents of 0.2%
and 1.8% were cited in the study of Omari et al. [16]. The low
moisture content is desirable because of less susceptibility of the
seeds to destruction of microbes and lipase enzymes. Too much
water in seeds made them vulnerable to hydrolysis by lipase
enzyme. The actions of microbes and lipase may reduce the quality
of R. communis bio-oil produced since their activity results in the
production of free fatty acid which may elevate the level of acid
value making the bio-oil unfit for biodiesel making [16]. The
R. communis seeds have a moderate ash content of 3.6 ± 0.4% which
was comparable to previously reported value of 3.8% [17]. Based on
the work of Veeresh and Narayana [18], the castor seed cake
(biomass) has a heating value of 20.1 MJ kg
1, volatile combustible
matter of 90.2% and fixed carbon of 9.0%.
Table 1
Characteristics of R. communis seeds.
Characteristic Value
Moisture content (fresh)a 7.6 ± 1.6
Moisture content (dry)b 1.9 ± 0.0
Ash content 3.6 ± 0.4
a
As received.
b
Dry basis.
The heating value which is a very important factor affecting
utilization of any material as a fuel is affected by the moisture and
ash content. Moisture and insoluble compounds act as a heat sink
that lowers the combustion efficiency [19]. Thus, the moisture and
ash contents of R. communis seeds presented in Table 1 were
desirable since low moisture, and ash contents increased the
heating value that made it more desirable as fuel [20].
3.2. Parametric study on the range of values of the variables
Preliminary experiments were conducted to determine the
levels and range of reaction variables which were eventually used
during the optimization of bio-oil yield of R. communis seeds. The
independent variables studied were HB ratio (7:1e11:1), extraction
time (20e40 min) and resonance amplitude (40e60 mm). Percent
R. communis bio-oil yield was taken as the dependent variable.
Fig. 2a shows the bio-oil yield using 15 g of biomass with a varied
HB ratio while the extraction time and resonance amplitude was
taken constant at 30 min and 50 mm, respectively.
It can be observed from the graph that the peak was found at 9:1
HB ratio with 41.5% bio-oil yield. It also showed that the HB ratio
affected the yield of R. communis since the yield increased with the
increased of the HB ratio. Similar results had been shown from the
preliminary study conducted by Dasari and Goud [15] in which they
obtained the highest yield for R. communis bio-oil extraction at the
ratio of 9:1. Hence, the 9:1 HB ratio was applied as the center (level
0) in the next stage of the parametric study. The observed decrease
of the bio-oil yield with higher HB ratio could be attributed to
excess amount of hexane that might degrade the biomass with
temperature during extraction [21]. More concentration of solvent
was also found to have inhibitory effect of the reaction rate that
usually led to a decrease in extraction yield [22].
On the next stage, the extraction time was varied from 20 to
40 min at 5 min interval while the HB ratio was fixed at 9:1 and
resonance amplitude at 50 mm. Fig. 2b shows the bio-oil yield from
15 g R. communis seeds at different extraction time. It can be seen
that 30 min extraction time was the peak with 41.9% bio-oil yield.
The result indicated that there was a continuous increase in the bio-
oil yield and the ultrasonication process was more effective from 20
to 30 min. The research of Balachandran et al. [23] confirmed that
ultrasound continued to be effective even during the later stage of
extraction through an enhancement to the internal diffusivity. In
any case, based on the results so far, 30 min was considered as the
center of the level of the extraction time (level 0) in the next stage
of the parametric study. It could also be observed that further in-
crease in extraction time resulted in a decrease of bio-oil yield
because, at longer duration, a significant amount of solvent is lost
 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453 447

Fig. 2. Bio-oil yield of R. communis seeds at different parameters: (a) varied HB ratio (Extraction time: 30 min, Resonance amplitude: 50 mm); (b) varied extraction time (HB ratio:
9:1, Resonance amplitude: 50 mm); and (c) varied resonance amplitude (HB ratio: 9:1, Extraction time: 30 min).

due to evaporation and the amount of solvent left is not enough for current results, the resonance amplitude of 50 mm was chosen as
efficient extraction [4]. Another possible reason is the degradation the center of the levels of the resonance amplitude (level 0).
of oil due to long extraction time and prolonged exposure to heat Based on the preliminary experiments, HB ratio of 9:1 (mL g
1),
[21]. an extraction time of 30 min, and a resonance amplitude of 50 mm
On the last stage of the parametric study, the resonance were chosen as the central conditions for the optimization study.
amplitude was varied from 40 to 60 mm at 5 mm interval while the Optimization study further investigated the effects of the reaction
HB ratio was fixed at 9:1 and extraction time at 30 min. Fig. 2c il-
lustrates the bio-oil yield at different resonance amplitude. Refer- Table 2
ring to the graph, when the resonance amplitude was increased Experimental range and levels of independent variables.
from 40 to 50 mm, the yield of R. communis seeds bio-oil was
Variable Coded level
increased from 24.5 to 38.9%. However, increasing the resonance
amplitude further to 60 mm resulted in a decrease in bio-oil yield
2
1 0 1 2

from R. communis seeds. This may be a result of cavitational heating n-hexane to biomass ratio (mL g
1) 7 8 9 10 11
as concluded by Tian et al. [21]. The peak was found at 50 mm which Extraction time (min) 20 25 30 35 40
Resonance amplitude (mm) 40 45 50 55 60
was consistent with the study of Zhang et al. [24]. Based on the
448 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453

variables for maximum bio-oil yield. Table 2 shows the range of Table 4
independent variables and their coded levels that were derived Analysis of variance of response surface reduced quadratic model for percentage
bio-oil yield from R. Communis seeds.
after preliminary runs and were used in the experimental design.
Source Sum of df Mean F P-value
square
3.3. Optimization of bio-oil yield from Ricinus communis seeds square value Prob > F

Model 496.57 8 62.07 84.06 < 0.0001a

The 20 experimental runs and the results of the optimization A-HB ratio 82.81 1 82.81 112.15 < 0.0001a
study are shown in Table 3. The optimum yield of 42.5% was way B-Extraction time (min) 1.93 1 1.93 2.62 0.1340b
C-Resonance amp (mm) 3.82 1 3.82 5.18 0.0439a
higher than the reported yield of 35.5% by the study of Mbah et al.
AB 25.78 1 25.78 34.91 0.0001a
[25] which used leaching process in the extraction of bio-oil from BC 4.77 1 4.77 6.47 0.0273a
R. communis seeds. It was also comparable to the yield of 43.7% from A2 98.13 1 98.13 132.89 < 0.0001a
the same kind of seeds through Soxhlet extraction method in the B2 64.71 1 64.71 87.64 < 0.0001a
research of Omari et al. [16]. C2 329.13 1 329.13 445.73 < 0.0001a
Residual 8.12 11 0.74
Analysis of variance (ANOVA) was performed to evaluate the
Lack of fit 3.06 6 0.51 0.50 0.7855b
quality of the model. The regression coefficients and significant Pure error 5.06 5 1.01
probabilities of linear (A: HB ratio, B: extraction time and C: reso- Cor Total 504.69 19
nance amplitude), quadratic (A2, B2, and C2) and interaction effects R2 ¼ 0.9839
of process variables (AB and BC) are shown in Table 4. Probability a
¼ Significant.
values (p-value) of  0.05 had significant effects on the bio-oil yield b
¼ Not significant.
while values greater than 0.05 indicated that the model terms were
not significant. In this case, A, C, AB, BC, A2, B2 and C2 were signif-
icant model terms. According to the table, the p-value of the extraction time and resonance amplitude had positive coefficients,
generated reduced quadratic model of < 0.0001 was less than 0.05 when all taken singly, meaning that the increase of their values
which revealed that model fitness was significant. Moreover, the could independently increase the bio-oil yield up to certain limits.
“Lack of fit-value” of the model was not significant with a p-value of The increase of the numerical product of the interactive term BC
0.7855, implying that there was a 78.55% chance that the lack of fit could also increase the bio-oil yield while the increase in AB would
this large occurred due to noise. result in a decrease in yield. The negative coefficients of the
The reduced quadratic model equation obtained from the quadratic terms A2, B2 and C2 also implied that the increase in these
experimental data to predict the yield of bio-oil in terms of actual terms did not give any significant increase in the bio-oil yield and
factors is given in Eq. (2). would result otherwise if the increase was too high. Also, high
variable coefficients represented greater effects of the particular
% Bio
oil Yield ¼
612:25 þ 48:61A þ 5:47B þ 13:64C parameter in the variable under investigation [27]. The HB ratio had
the most influential effect on the bio-oil yield from R. communis

0:36AB þ 0:03BC
1:98A2
0:06B2 seeds. Since there are many significant variables (linear, quadratic

0:14C 2 (2) and interactive), the effects cannot be singly concluded. Hence, the
model equation can serve as best solution to predict the bio-oil
where, A, B, and C are actual variables of the predictors namely HB yield.
ratio, extraction time (min) and resonance amplitude (mm) Fig. 3 shows the graphical comparison between the actual and
respectively. The sign of the coefficients determined if a parameter predicted bio-oil yields of R. communis seeds. It is observed that the
has a positive or adverse effect on the percent bio-oil yield. Positive actual yield plotted is near the trend line representing the predicted
coefficients of the equation indicate an increase in the bio-oil yield bio-oil yield with the coefficient of determination (R2) of 0.9839.
once the value of the coefficient is increased. On the other hand, the This result supports the reasonable precision of the generated
negative coefficients signify otherwise [26]. The variables HB ratio, model equation.

Table 3
Experimental designs and bio-oil yield extracted from R. communis seeds.

Run HB ratio Extraction time (min) Resonance amplitude (mm) Yield (wt%)

1 9 30 50 42.5
2 9 30 40 25.7
3 9 20 50 35.2
4 8 25 45 30.8
5 9 30 50 41.1
6 10 35 55 34.4
7 10 35 45 32.6
8 11 30 50 38.4
9 7 30 50 28.7
10 8 35 55 34.1
11 9 30 50 40.2
12 9 30 50 41.2
13 9 30 60 28.3
14 9 40 50 34.9
15 9 30 50 42.5
16 10 25 55 37.8
17 8 35 45 31.5
18 8 25 55 30.0
19 10 25 45 38.7
20 9 30 50 40.3
 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453
Fig. 3. Graphical comparison between actual and predicted bio-oil yield.
3.4. Influence of the parameters on the bio-oil yield
Interactive AB and BC were significant model terms. Hence, they
were considered in the discussion of the parametric influence on
the bio-oil yield. The response surface plots and the contour plots
representing the effects between the independent and dependent
variables for different fixed parameters are shown in Fig. 4.
The effect of varying HB ratio and extraction time at constant
resonance amplitude of 50 mm on bio-oil yield is shown in Fig. 4a.
The response surface indicated that the percentage yield of bio-oil
increases with HB ratio at 8 to 10 (v w
1). It was also seen that the
effect of HB ratio was more noticeable than the extraction time. This
suggested that the HB ratio was more influential variable affecting
the bio-oil yield from R. communis seeds. This was evident from its
numerical coefficient in Eq. (2) and its p-value in the ANOVA. It
denoted that high HB ratio would result in high bio-oil yield of
R. communis seed. This observation was consistent with the mass
transfer principle, where the driving force during the mass transfer
was the concentration gradient between the solid and the bulk of
the liquid, which increased when a higher solvent to biomass ratio
was used [21]. On the other hand, there was only less prominent
increase in the bio-oil yield as extraction time increased. Therefore,
similar to the study of Zhang et al. [24], a longer extraction time was
unnecessary once the maximum extraction yield was achieved.
The effect of time and resonance amplitude at constant HB ratio
at 9:1 (v w
1) is depicted in Fig. 4b. It was found out that an in-
crease of bio-oil yield can be observed with resonance amplitude
between 40 and 50 mm. On the other hand, it showed a decreasing
trend with resonance amplitude higher than 50 mm. This finding
confirmed the ANOVA results wherein the resonance amplitude
had a significant p-value. Thus, the resonance amplitude was more
influential variable than the extraction time. Similar results were
obtained for the bio-oil from rice bran [28] and flaxseed [24] using
UASE. This may be due to that fact that higher amplitude may
damage more cell walls thereby releasing more crude bio-oil to the
solvent [29]. Furthermore, UASE had been reported to increase bio-
oil extraction through the action of accelerating the rehydration or
swelling of cells accompanied by the fragmentation of the tissue
449
matrix [30]. This involved the mass transfer and penetration of the
solvent into the cell promoting absorption of cell contents into the
solvent [31].
In this study, increasing the resonance amplitude higher than
50 mm, and at longer extraction time decreased the percent bio-oil
yield. A similar result was also observed by Ido et al. [26]. The
decrease in bio-oil yield was because during the extraction process,
higher resonance amplitude and extraction time raised the tem-
perature rapidly. At such, higher temperatures vaporized the solvent
rapidly and not enough of it was available to extract bio-oil [32].
The overall interactions of each of the variables on the three-
dimensional surface plots demonstrated that the influence from
greatest to least was HB ratio > resonance amplitude > extraction
time. This order was exemplified from the p-values in the ANOVA.
3.5. Validation of the optimum conditions generated by CCD
The CCD of RSM suggested solutions which generated optimum
conditions. The solution with the highest desirability (0.98) was
chosen and was validated in actual runs. The predicted optimum
bio-oil yield was 42.0% at optimum conditions of 10:1 HB ratio,
25 min extraction time, and 50 mm resonance amplitude. The re-
sults of the actual runs were compared to the CCD prediction as
presented in Table 5. Three verification runs were conducted and
obtained an actual bio-oil yield of 42.2 ± 0.4%. The results of the
actual runs were acceptable since the recorded percent errors
(0.6e1.2%) were less than the 5% acceptable error. Thus, this result
attested to the reliability of the generated model for optimum and
effective bio-oil extraction.
3.6. Product analysis and characterization
Characteristics of UASE-extracted bio-oil from R. communis
seeds as compared with other studies is presented in Table 6. The
pale yellow bio-oil from R. communis seeds obtained in this study
was similar with the result of Dasari and Goud [15]. Density was
found to be lower value of 0.9 g mL
1 for the UASE-extracted
R. communis seed bio-oil as compared to other methods of
extraction and other related studies (Table 6). The result was
desirable considering that high-density fuel leads to incomplete
combustion and particulate matter emissions [21].
Its kinematic viscosity of 18.7 cSt was lower than the reported
value of Dasari and Goud [15] and Tunio et al. [33] having 230 and
196 cSt, respectively. It was also a little lower than the kinematic
viscosity of 29.7 and 6.6 cSt found in the study of Demirbas [34] and
Kılıç et al. [10], respectively. It was assumed that UASE significantly
reduced the viscosity of R. communis seed produced bio-oil. The
reduction of the viscosity of bio-oils is important and vital for en-
gine lubrication and better performance [35].
Higher heating value (HHV) is another important parameter
considered in this study. The HHV of 29.5 MJ kg
1 was quite lower
than the findings of other studies (Table 6). According to Arazo et al.
[27], a highly desirable bio-oil should exhibit HHA. It was assumed
that a decrease in heat content of UASE-extracted bio-oil from
R. communis seeds was the result of the greater unsaturation in the
bio-oil [34].
FTIR spectrum (4500e500 cm
1) of R. communis seed bio-oil
extracted via UASE revealed major functional absorption bands
(Fig. 5). The broad band peaking at 3340.7 cm
1 represented OeH
stretching vibration for hydrogen-bonded OH groups. The hydrox-
yl groups present in the R. communis seed produced bio-oil could
generate methyl groups (eOeCH3) through methyl trans-
esterification [36]. The only carbonyl group present occurred at
band wavenumber 1745.0 cm
1 for C]O stretching of triglyceride
ester linkage and 1650.6 cm
1 for C]C stretching vibration [37].
450 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453

Fig. 4. Response surface plot showing the (a) 3D effect and contour plot of extraction time and HB ratio; (b) 3D effect and contour plot of extraction time and resonance amplitude
on the bio-oil yield.

Table 5
Optimization and validation of bio-oil yield from R. communis seeds via UASE.

Experiment Operating variable Bio-oil yield (%) Error (%)

HB ratio (v w
1) Extraction time (min) Resonance amplitude (mm)

CCD 10 25 50 42.0
Validationa 10 25 50 42.2 ± 0.4 0.6 ± 1.2

Optimization criteria: in range e HB ratio and resonance amplitude; minimize e extraction time; maximize e yield.
a
0.98 desirability.

Table 6
Comparison of the characteristics of UASE-extracted bio-oil from R. communis seeds to other studies.

Property R. communis seed bio-oil R. communis seed R. communis seed bio-oil R. communis R. communis
via UASE at optimum bio-oil extracted extracted through seed bio-oilc seed bio-oild
conditions (this study) via Soxhleta mechanical screwb
Expellerb

Color Pale yellow Pale yellow Colorless e e

Density (g mL
1) 0.90 0.98 0.96 e 0.97
Kinematic viscosity at 40  C (cSt) 18.7 230 196 29.7 6.6
Higher heating value (MJ kg
1) 29.5 38.2 35.2 37.4 40.8

In literatures c and d, the extraction process were not stated;
no reported value.
a
Dasari and Goud [15].
b
Tunio et al. [33].
c
Demirbas [34].
d
Kılıç et al. [10].
 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453 451

Fig. 5. FTIR spectrum of R. communis seed bio-oil produced via UASE.

Fig. 6. GCeMS chromatogram of R. communis seed bio-oil via UASE.

Table 7
Other well-defined peaks were those for CeH stretching Fatty acid profile of R. communis seed bio-oil via UASE.
(3000.8 cm
1), CeH stretching (CH2) (2990e2875 cm
1), CH
bending (1450.8 cm
1), CH3 bending (1360.5 cm
1), CN stretching Fatty acid composition Fatty acid relative content (%)

(1150.9 cm
1), CeO stretching (1075.9 cm
1), CeH out of plane vi- Total saturated fatty acids (11.3%)
bration (828.9 cm
1), and CeCl stretch (695.0 cm
1). The CeN group Palmitic acid (C16:0) 2.2
Stearic acid (C18:0) 2.4
detected with the stretch mode of vibration may indicate the
Oleic acid (C18:1n9c) 6.7
presence of undesirable contaminant [38]. Total unsaturated fatty acids (87.8%)
GCeMS was used to identify and measure the composition of Linoleic acid (C18: 2n6c) 8.8
fatty acids present in bio-oil. Fig. 6 shows the GCeMS chromato- Linolenic acid (C18:3n3) 0.7
Arachidic acid (C20:0) 0.1
gram of bio-oil while the identified fatty acids are presented in
Eicosenoic acid (C20:1) 0.9
Table 7. Eicosadienoic acid (C20:2) 0.1
The major saturated fatty acids (SFA) are palmitic (2.2%), stearic Eicosapentaenoic acid (C20:5n3) 0.1
(2.4%) and oleic (6.7%). On the other hand, high amount of Ricinoleic acid (C18:1) 77.1
452 C.L. Palconite et al. / Sustainable Environment Research 28 (2018) 444e453

60
52.6

42.2
40
Bio-oil Yield (%)
29.6

20

0
(a) UASE (b) Soxhlet (c) Magnetic
Extraction Stirring

Extraction Method
(a) optimum condition: 10:1 HB ratio, 30 min extraction time, 50 µm resonance amplitude,
15 g biomass;
(b) 20:1 n-hexane to biomass ratio, 6 h extraction time, 70 °C, 15 g biomass;
(c) 10:1 n-hexane to biomass ratio, 3 h extraction time, 700 rpm, ambient
temperature, 5 g biomass
Fig. 7. Comparison of bio-oil yields of UASE, Soxhlet extraction and extraction via magnetic stirring.

unsaturated fatty acids (UFA) comprised of linoleic (8.8%), linolenic
(0.7%), arachidic (0.1%), eicosenoic (0.9%), eicosadienoic (0.1%),
eicosapentaenoic (0.1%), and the most abundant ricinoleic (77.1%).
R. communis seed produced bio-oil is unique among vegetable oils
and its uniqueness is derived from the presence of hydroxylated 12-
hydroxy, 9-octadecenoic acid, known familiarly as ricinoleic acid.
No other vegetable oil contains this type of hydroxyl fatty acid
[39,40].
Moreover, it can be observed from the table that UFA content of
the extracted bio-oil from R. communis seeds was 87.8% while
there was a relatively low content of SFAs of 11.3%. SFA tend to
have a higher melting point than the UFA. Thus, high amount of
UFA does not have adverse effects on biodiesel during cold
weather engine operation. Moreover, unsaturated biodiesel fuels
emit lower hydrocarbon, carbon oxide and smoke emissions as
compared to highly saturated biodiesel fuel [40]. The downside of
UFA is that these are often more prone to oxidation and rancid-
ification than the saturated counterparts. Biodiesel made from
abundant UFA require an oxidative stabilizer to be used safely as
fuel [41].
3.7. Comparison of UASE, Soxhlet and magnetic stirring extraction
The percent bio-oil yield extracted via UASE was compared at
different extraction methods through Soxhlet extraction and
magnetic stirring (Fig. 7). It shows that the yield was apparently
higher via use of the ultrasonic method (42.2%) compared with the
magnetic stirring mechanism (29.5%). This was because of the fact
that the chemical reaction is accelerated with the help of cavitation
phenomenon. As a result of this, improvement in mass transfer of
the mixture and enhancement in rate of reaction between mole-
cules took place [42]. The results also revealed that the yield of
Soxhlet extraction process was 52.6%, and way higher than the
UASE-extracted bio-oil. Undeniably, Soxhlet extraction is the best
when compared with other methods, because the process is
continuous and there is complete bio-oil recovery. However, it
should be noted that the UASE showed a promising result since it
used only 150 mL of solvent in 30 min, which was way lower than
300 mL in 6 h through Soxhlet extraction method with a compa-
rable bio-oil yield.
4. Conclusions
In this study, the bio-oil yield through UASE reached 42.2 ± 0.4%
at optimum conditions of 10:1 HB ratio, 25 min extraction time and
50 mm resonance amplitude using the CCD of the RSM. The char-
acterization of bio-oil resulted to a density of 0.9 g mL
1, kinematic
viscosity at 40  C of 18.7 cSt and higher heating value of
29.5 MJ kg
1. The FTIR analysis revealed the presence of carbonyl
group (C]O and C]C), a desirable ester functional groups con-
taining esters for biodiesel production. UFAs dominated the bio-oil
which include linoleic (8.1%), linolenic (0.7%), arachidic (0.1%),
eicosenoic (0.9%), eicosadienoic (0.1%), eicosapentaenoic (0.1%) and
ricinoleic (77.1%) with low amount of SFAs such as palmitic (2.2%),
stearic (2.4%) and oleic (6.7%).
Acknowledgement
The authors would like to express their sincere appreciation to
the faculty and staff of the College of Engineering and Technology of
the University of Science and Technology of Southern Philippines -
Claveria for the valuable assistance and support.
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