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Energy for Sustainable Development 23 (2014) 110114

Contents lists available at ScienceDirect

Energy for Sustainable Development

Optimization of biodiesel production from waste lard by a two-step


transesterication process under mild conditions
Ioannis Sarantopoulos a, Efthalia Chatzisymeon b, Spyros Foteinis a, Theocharis Tsoutsos a,
a
b

School of Environmental Engineering, Technical University of Crete, Polytechneioupolis, GR-73100 Chania, Greece
Institute for Infrastructure and Environment, School of Engineering, The University of Edinburgh, Edinburgh EH9 3JL, United Kingdom

a r t i c l e

i n f o

Article history:
Received 17 September 2013
Revised 5 July 2014
Accepted 12 August 2014
Available online xxxx
Keywords:
Waste lard
Biodiesel
Two-step transesterication
Factorial design

a b s t r a c t
The aim of this work is to investigate a two-step homogenous catalyzed waste lard transesterication reaction for
low cost biodiesel production. For this purpose, two 23 full factorial design of experiments was applied. Six variables, namely esterication time (60120 min), H2SO4 concentration (2040 wt.%), MeOH:FFA (15:123:1),
transesterication time (3060 min), KOH concentration (12 wt.%), and MeOH:triglycerides (6:19:1), that
typically affect the production process were studied. The esterication step is signicantly affected mostly by
the reaction time and the MeOH:FFA ratio value. Specically, their increase brings a reduction of the FFA acidity.
Likewise, the transesterication step is positively affected primarily by three independent variables, namely reaction time, KOH concentration and MeOH:triglyceride ratio. Furthermore two empirical models describing evolution of the two-step transesterication reaction were developed. They can become useful tools for further
scaling-up the process by predicting its reaction yield within a 95% of condence level.
2014 International Energy Initiative. Published by Elsevier Ltd. All rights reserved.

Introduction
Recently, biodiesel has received increasing attention worldwide due
to the existing sustainable energy policy framework. Moreover, it
presents unique advantages such as being biodegradable, suitable for
domestic production and non-toxic (Ouachab and Tsoutsos, 2013;
Tsoutsos et al., 2010; van Stralen et al., 2013). On the other hand, in
comparison to petrodiesel, biodiesel has 12% lower energy content,
higher NOx emissions in the range of 1014% during combustion,
needs higher injector pressure, lower oxidation stability and higher
production cost (Atabani et al., 2012). In recent years, systematic efforts have been made to produce biodiesel through the utilization
of waste oils, such as used cooking oils (Wyse-Mason and Beckles,
2012; Math et al., 2010; Talebian-Kiakalaieh et al., 2013; Lee et al.,
2002), animal fats (Kleinov et al., 2011; Berrios et al., 2009), vegetable and rened oil products (Alamu et al., 2007; Che et al., 2012;
Sarantopoulos et al., 2009), which are used to produce a costcompetitive product.
The free fatty acid (FFA) contents of crude vegetable oils, used
cooking oils, animal fats and grease range from 0.3 to 0.7, 2 to 7, 5 to
30 and 40 to 100%, respectively. However, the high amounts of FFAs in
these low cost biodiesel feedstocks make them unsuitable for base-

Corresponding author. Tel.: +30 2821037825.


E-mail address: theocharis.tsoutsos@enveng.tuc.gr (T. Tsoutsos).

catalyzed transesterication, due to the soap formation between FFAs


and the base catalyst (Vicente et al., 2004). The presence of soap can
prevent the separation of biodiesel from glycerol and cause the formation of emulsions (Endalew et al., 2011; Marchetti et al., 2007). This
leads to an increase in the product viscosity and in the joint cost of
separation and purication. The saponication can be avoided through
the acid-catalyzed process because it can simultaneously transform
FFAs and triglycerides (TGs) to FAME by esterication, before the
transesterication process. This method is known as two-step acid
base catalyzed transesterication and is designed as an alternative
process for biodiesel production from oils with high FFA content
(Talebian-Kiakalaieh et al., 2013).
Waste lard is a by-product of hog industry with limited other uses
and low market price. Availability is already high, the hog industry in
Greece consumes about 120 M kg of pork (GMRDF, 2014), and if proper
waste management schemes will be implemented, an additional increase in the production is expected. Waste lard is a by-product of
pork meat industry and until now is used as feedstock in the animal
feed industry. However, the risk of animal disease had a detrimental
effect on the use and the prices of these animal origin raw materials.
Therefore, nowadays biodiesel industry can provide a stable and secure
alternative market for these raw materials, which can contribute also to
the sustainability of the biodiesel fuel use.
Hence, the aim of this work is to optimize the two-step homogeneous catalyzed transesterication, with the ultimate purpose of
producing biodiesel from a low-cost feedstock, such as the waste lard,
under mild operating conditions. For this purpose, a factorial design of

http://dx.doi.org/10.1016/j.esd.2014.08.005
0973-0826/ 2014 International Energy Initiative. Published by Elsevier Ltd. All rights reserved.

I. Sarantopoulos et al. / Energy for Sustainable Development 23 (2014) 110114

experiments (DOE) methodology was utilized and six variables that


typically affect the two-step homogeneous catalyzed transesterication
reaction were investigated.

Materials and methods


Raw material
Dehydrated lard was provided by a meat industry company located
in Rethymnon, Crete, Greece. The feedstock was used as it was obtained
without any further rening treatment prior to the experimental
process. This waste lard had an initial acid value of 13.06 mg KOH g1
corresponding to a FFA level of 6.56% and its fatty acid composition, as
determined by gas chromatography, is shown at Table 1. It should be
noted that in the specic industry vegetable oils are used as animal fat
substitutes and are incorporated into the preparation process of meatbased products to improve their avor and their dietary content
(Domazakis, 2010). This is reected in lards' fatty acid content presented in Table 1. For example, C18:2 are 21.7% in lard, which is relatively
high for animal fats. This may be attributed to the presence of various
vegetable oils, with a C18:2 content higher than 20% (Mamat et al.,
2005), in the lard products.
Two-step batch process
Acid esterication step
Feedstock with acidity above 1% cannot be converted efciently to
biodiesel with the conventional alkaline process. If FFA content is
above this limit, soap formation decreases catalyst activity and inhibits
the reaction rate and time for completion. Moreover, water formation
is causing more soap formation by hydrolyzing the triglycerides to
free fatty acids. Increased soap content in reaction mixtures also
decelerates glycerol separation and increases settling time for adequate
glycerol removal.
Esterication catalyzed by homogeneous acid catalyst is a common
way for converting FFA to esters. After this pretreatment step, the
reaction mixture has a low acidity and can be followed by a common
alkaline transesterication so as to convert triglycerides to the desired
nal product. For the experimental tests, 50 g of raw material were loaded into a 500 mL spherical three-neck reaction ask, which was immersed in hot water bath at 50 1 C. The reactor was preserved
under a water-cooled condenser for recovering methanol vapors and
under continual magnetic stirring.

Transesterication method
Transesterication tests were conducted in a 500 mL spherical
three-neck reaction ask, which was immersed in hot water bath at
50 1 C and 50 g of the esteried lard and the appropriate KOH
amount were loaded in it. The reactor was preserved under a watercooled condenser for recovering methanol vapors and under continual
magnetic stirring. Periodically (at 30 and 60 min) samples of 5 mL
were collected and their methyl-ester content was measured.
Table 1
Fatty acid prole of waste lard raw material.
Fatty acid

Class

wt.%

Myristic acid
Palmitic acid
Palmitoleic acid
Stearic acid
Oleic acid
Linoleic acid
Linolenic acid
Others

C14:0
C16:0
C16:1
C18:0
C18:1
C18:2
C18:3

1.0
21.1
1.5
11.5
40.1
21.7
1.5
2.3

111

Biodiesel quality determination


After the two-step transesterication process and optimal operating
parameters, the produced biodiesel was analyzed in terms of its quality
parameters. Therefore, this biodiesel sample was further puried to
achieve highest quality characteristics. For this purpose crude biodiesel
was centrifuged at 4000 rpm for 5 min and the supernatant glycerol was
removed. Afterwards biodiesel was submitted to wet washing with distilled water of ~ 50 C to remove any impurities. Further purication
took place with a commercial dry washing process with magnesium silicate as solid absorbent. A 2 wt.% ratio of magnesium silicate to biodiesel
was stirred for 15 min at 60 C. Finally, biodiesel was ltered through
1 m lters to remove magnesium silicate remnants and other solid
impurities present in the fuel.
Analytical measurements
Gas chromatography (GC) was performed on a Shimadzu (GC-17A),
equipped with ame ionization detector (FID) and mounted with a
Mega Biodiesel 103 0.2 mm 0.25 m 30 m column. Helium was
used as carrier gas. Injector and detector temperatures remained constant at 300 C. Temperature program of the column oven started at
200 C, then to 240 C at a rate of 5 C/min, held at 240 C for 5 min,
to 260 at a rate of 5 C/min and nally, held at 260 C for 5 min.
The acid value of the lard, expressed as mg KOH required to neutralize 1 g of biodiesel, was obtained according to the ofcial standard
method for the determination of acid value and acidity in vegetable
and animal oils and fats of the European Committee for Standardization
(EN 660:1999).
Total and free glycerol and mono-, di- and triglyceride contents were
measured by gas chromatography according to EN 14105 analytical
standard. Analysis was performed in GC-FID SRI 8610C instrument
mounted with Restek MXT 500 biodiesel gas chromatography column
with an internal diameter of 0.53 mm, a lm thickness of 0.15 m and
a length of 15 m, with helium as carrier gas.
Ester content, linolenic acid methylester and polyunsaturated (N_4
double bonds) methyl ester contents were measured according to EN
14103 analytical standard procedure. Analysis was performed in a
Shimadzu GC-FID 17A gas chromatograph, mounted with gas chromatography column Mega Biodiesel 103 with an internal diameter of
0.32 mm, a lm thickness of 0.25 m and a length of 30 m.
The density at 15 C was measured with a density meter instrument
Anton Paar DMA 38, calibrated and performed according to ASTM
D4052.
Water evaporation from the nal biodiesel product was performed
on a rotary evaporator Heidolph Laboretta 4011-digital.
Factorial design and analysis of the experiments
A statistical approach was chosen, based on a factorial experimental
design, to infer about the effects of the variables on process performance. Six independent variables (X1 to X6) that typically affect the
two-step biodiesel production process efciency were taken into
account. Each received a high (+) and a low () value, as seen in
Table 2
Independent variables of the two different 23 factorial designs of experiments for each
process step.
Esterication step
Value level

+
0

Transesterication step

X1

X2

X3

X4

X5

X6

Time,
min

H2SO4, wt %
of FFAs

MeOH:
FFA

Time,
min

KOH, wt %
of oils

MeOH:
triglyceride

60
120
90

20
40
30

15:1
23:1
19:1

30
60
45

1
2
1.5

6:1
9:1
7.5:1

I. Sarantopoulos et al. / Energy for Sustainable Development 23 (2014) 110114

Table 2. Analytically, biodiesel production process performance was


evaluated in terms of % transformation yield (response factors Y1 and
Y2 for the esterication and transesterication steps, respectively). For
this purpose, two individual 23 full factorial designs were performed.
The design matrix of the experiments and their statistical analysis
were performed by means of the software package Minitab 16. Analysis
of the response factors Y1 and Y2 involves the estimation of the average
effect, the main effects of each individual variable, as well as their two
and higher order interaction effects (Box et al., 1978). The present
study was done for a condence interval of 95%. An estimate of the
standard error was obtained by repeating all the experimental runs of
the 23 factorial designs. Although the entire reproducibility of the factorial design is time-consuming, it is the safest practice to accurately estimate the standard error of the experimental values. Therefore, all
experimental runs were performed in duplicates so as to minimize
experimental error and increase model accuracy. In this case, identication of the important effects was carried out by computing the corresponding p-values of the experimental data. If the p-value is 0.05
then that effect is not statistically signicant, at the 95% condence
level. Likewise, the alpha (a-level) value corresponds to the maximum
acceptable level of risk for rejecting a true null hypothesis. Hence, all
effects greater than the a-level value, in absolute values, are considered
as statistically signicant. Duplicate experiments were also conducted
at the center point of the factorial design (at zero level) to further ensure
the repeatability of the process.
Results and discussion
Effect of operating parameters on process efciency
Table 2 shows the selected parameters and their respective high and
low values for these two DOEs. These parameters were reaction time
(X1), H2SO4 concentration (X2) and MeOH-to-FFA ratio (X3), for the
acid esterication step. The response was the percentage of the acidity
reduction of the FFAs (Y1), according to Eq. (1).
Y1

AV in AV f
 100
AV in

Where AVin is the initial acidity of the waste lard (mg KOH/g), and
AVf is the acidity (mg KOH/g) of the nal mixture after the end of the
esterication step.
During the transesterication reaction process variables under
investigation were reaction time (X4), KOH concentration (X5) and
MeOH-to-oil ratio (X6). At this point it should be mentioned that the
average molecular weight of the lard was estimated based on its FFA's
prole shown at Table 1. In this factorial design the response factor is
the percentage of methyl ester production (Y2), as determined by gas
chromatography after sample purication. The above parameters were
chosen, as these are typical parameters that affect biodiesel process
production. However, the effect of all the above parameters and their
combination has not been investigated yet for the two-step biodiesel
production form waste lard. Table 3 shows all the experimental runs
performed in duplicate of the two 23 factorial designs.
A key element in the factorial design statistical procedure is the determination of the signicance of the estimated effects. A very useful
pictorial presentation of the estimated effects and their statistical importance can be accomplished using the Pareto chart of the effects.
This chart displays the absolute value of the effects in a bar chart and
draws a reference line on the chart. Any effect that extends past this reference line (corresponding to -level value) is potentially important.
On the other hand, all other effects whose values are lower than the
reference can be attributed to random statistical error. The Pareto
chart of the effects for the esterication step (response Y1) is shown in
Fig. 1. There are ve effects that are greater than the reference decision
threshold. Among them, the two most signicant ones are the time (1)

Table 3
Design matrix of the two individual 23 factorial experimental designs and the observed
response factors Y1 and Y2, during the esterication and the transesterication step
processes, respectively.
Esterication step

Transesterication step

Run order

X1

X2

X3

Y1

Run order

X4

X5

X6

Y2

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18

+
0

0
+

+
+

+
+

+
+
0
+
0

+
+
+

+
+

+
0

+
+
+

92.41
84.43
77.84
92.33
89.11
75.23
89.42
86.27
89.88
94.20
88.96
72.47
86.43
88.57
71.70
93.63
88.50
83.74

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

+
+
+
+
+
+
+

+
+
+
+

+
+

+
+
+

+
+

76.4
84.3
77.2
93.9
97.2
93.9
83.2
91.4
90.8
86.2
91.2
96.8
86.9
91
85.3
84.1

and the MeOH:FFA ratio (3), revealing a positive effect on treatment


efciency. This means that an increase in their level brings an increase
on esterication process performance, thus increasing the acidity reduction percentage of the FFAs. Alternatively, it is concluded that the esterication is favored at increased treatment time (i.e. 120 min) and high
MeOH:FFA ratios, such as 23:1. The variable of H2SO4 concentration affects positively but not as high as the other two variables esterication
efciency. Moreover, interactions of X1X3 and X1X2 affect negatively Y1
the response value. The three latter variables yield a substantially
lower effect on esterication efciency if compared with the X1 and X3
effects in absolute values. Hence, research interest should be further focused on the increase of the variables of treatment time and MeOH:FFA
ratio. Based on the Pareto chart it is derived that the duplicate runs 4
and 10 (shown at Table 3) reveal the optimal operating conditions for
the esterication as these were conducted at elevated treatment time,
MeOH:FFA ratio and H2SO4 concentration, which effects favor process
performance.
The Pareto chart of the effects for the transesterication step (response Y2) is shown in Fig. 2. It is observed that six effects are greater
than the reference decision threshold. The effects of the three variables
4, X5 and X6 are the most signicant, affecting positively the efciency
of the transesterication reaction. Besides, the effects of the two-order

a-level
A
C
AC

Term

112

B
AB

Factor Name
A
B
C

ABC
BC

10

15

20

X1
X2
X3

25

Standardized Effect
Fig. 1. Pareto chart of the effects for the esterication step for the full 23 factorial design.
White bars: positive effects; black bars: negative effects.

I. Sarantopoulos et al. / Energy for Sustainable Development 23 (2014) 110114


a-level

where Y1 is the FFA acidity reduction percentage (%), Xi are the transformed forms of the independent variables according to:

C
B

Xi

Term

A
AB

Factor Name
BC

A
B
C

ABC
5

10

15

20

Z high Z low
2
Z high Z low
2

Zi

and Zi are the original (untransformed) values of the variables.


The coefcients that appear in Eq. (2) are half the calculated effects,
since a change of X = 1 to X = 1 is a change of two units along the X
axis. The model predicts a linear dependency of the acidity reduction on
the operating variables. Therefore, Eq. (2) can be transformed to give Y1
prole as follows:

AC

113

X4
X5
X6

25

Y 2 49:2269 0:1627t 1:1339MeOH

Standardized Effect
Fig. 2. Pareto chart of the effects for the transesterication step for the full 23 factorial design. White bars: positive effects; black bars: negative effects.

interaction variables are statistically signicant but undoubtedly lower


than the effect of the independent variables, namely treatment time
(X4), KOH wt % (X5) and MeOH:triglyceride ratio (6). Hence, it is concluded that the transesterication step is favored at the increased values
of these variables (i.e. 60 min, 2% wt KOH, and MeOH:oil ratio equal to
9:1), which have been applied at runs 5 and 12 of the transesterication
factorial design.
Optimization of process efciency
The aforementioned statistical analysis allows inferring about the
variables and their interactions that are statistically signicant. What
is more, empirical models describing the reaction evolution in function
with the signicant effects can be developed. Based on these models optimal operating yields can be obtained during the two-step biodiesel
production process. The main effects of the independent variables and
their two and higher order interactions of the two factorial designs of
experiments are shown at Table 4.
Based on the variables and interactions that are statistically signicant, a model describing the experimental response Y1 for esterication
performance was constructed as follows:

Y 1 85:412

9:761
9:071
X1
X3
2
2

Table 4
Average and main effects of the independent variables and their two and higher order interactions of the two 23 factorial designs on the response factors Y1 and Y2.
Esterication step
Effect
Average effect
Main effects
X1
X2
X3
Two-factor interactions
X 1 X2
X 1 X3
X 2 X3
Three-factor interactions
X 1 X2 X 3
-level

Transesterication step
Value of
effect Y1
85.412
9.761
3.054
9.071
1.839
3.371
0.264
0.706
2.26

Effect
Average effect
Main effects
X4
X5
X6
Two-factor interactions
X 4 X5
X 4 X6
X 5 X6
Three-factor interactions
X 4 X5 X6
-level

Value of
effect Y2
88.1125
6.65
6.8
6.975
1.775
1.55
0.9
0.225
2.31

Where t is the reaction time (min), and MeOH is the MeOH:FFA


ratio.
Similarly, experimental models were developed for the
transesterication step, thus the response Y2 can be estimated by utilizing to Eq. (5), which is based on the most statistical signicant effects
according to Fig. 2.

Y 2 88:1125

6:65
6:8
6:975
X4
X
X6
2
2 5
2

where Y2 is the methyl-ester production percentage (%), Xi are the


transformed forms of the variables.
Based on Eq. (3), Y2 is transformed into the following empirical
model (6) that describes the formation of methyl-ester (Y2) during
the transesterication reaction step.
Y 2 50:5 0:2217t 6:8Cat 2:325MeOH

Where t is the reaction time (min), Cat is the KOH catalyst concentration (% wt), and MeOH is the CH3OH:triglycerides ratio. Hence, the
empirical models (4) and (6) can be a useful tool to further scale-up
the two-step biodiesel production process, within the operating points
utilized in these two full factorial designs of experiments.
Quality control
The development of biodiesel's industrial production was triggered
by extensive eld tests in running engines and vehicles so as to
determine the limits of various fuel properties. In Europe, the quality
of biodiesel is assessed under the provisions and the requirements of
quality standard EN 14214 that should be met from each biodiesel
sample so as to be a suitable substitute fossil diesel in transport sector.
Quality control was performed for the biodiesel produced under the optimum process conditions specied above (i.e. runs 4, 10 and 5, 12 for the
esterication and transesterication step, respectively), based on the
European Standard EN 14214:2003. Density at 15 C, acidity number,
methyl ester content, content of monoglycerides (MGs), diglycerides
Table 5
Quality control of produced biodiesel.
Properties

Produced biodiesel

Limits

Methyl ester content, %


Density at 15 C, kg/m3
Monoglyceride content, %
Diglyceride content, %
Triglyceride content, %
Free glycerol, %
Total glycerol, %
Acid value, mg KOH/g

99.4
877
0.0156
0.0005
0.0040
0.0031
0.0075
0.42

N96.5
860900
b0.8
b0.2
b0.2
b0.02
b0.25
b0.5

114

I. Sarantopoulos et al. / Energy for Sustainable Development 23 (2014) 110114

(DGs) and TGs and total and free glycerol, were determined and measured (Table 5). It should be noted that for further scaling up of biodiesel
production and its promotion in the market, all quality characteristics
specied by the European standard EN 14214 should be determined.
Conclusions
In this work, a two-step homogeneous catalyzed waste lard
transesterication reaction for biodiesel production was investigated.
For this purpose, a factorial design methodology of experiments was
utilized and six variables that typically affect the production process
were examined. The statistical signicant variables, their effects and
the higher order interaction effects on process efciency were identied. The main results drawn from this study are concluded below.
The esterication step is signicantly affected mostly by the reaction
time and the MeOH:FFA ratio value. Specically, an increase in their
level brings an increase in the FFA acidity reduction percentage. Likewise, the transesterication step is positively affected by three independent variables, namely reaction time, KOH concentration and MeOH:
triglyceride ratio, that typically affect its performance. Furthermore,
based on the statistically signicant variables, two empirical models describing evolution of the two-step biodiesel production process were
developed. This is of major importance since they are valuable tools to
further scale-up the process by computing its reaction yield. Finally, it
has to be noted that by applying the proposed two-step process, biodiesel, conformed to the European Community quality standards, can be
produced from a low cost feedstock, namely waste lard.
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