Optimization of direct conversion of wet algae to biodiesel under

supercritical methanol conditions

Prafulla D. Patil
a
, Veera Gnaneswar Gude
a
, Aravind Mannarswamy
a
, Shuguang Deng
a,
*
, Peter Cooke
b
,
Stuart Munson-McGee
a
, Isaac Rhodes
c
, Pete Lammers
c,1
, Nagamany Nirmalakhandan
d
a
Chemical Engineering Department, New Mexico State University, Las Cruces, NM 88003, USA
b
Electron Microscopy Lab, New Mexico State University, Las Cruces, NM 88003, USA
c
Chemistry and Biochemistry Department, New Mexico State University, Las Cruces, NM 88003, USA
d
Civil and Environmental Engineering Department, New Mexico State University, Las Cruces, NM 88003, USA
a r t i c l e i n f o
Article history:
Received 27 March 2010
Received in revised form 26 May 2010
Accepted 7 June 2010
Available online 29 June 2010
Keywords:
Biodiesel
Wet algae
Supercritical methanol
Response surface methodology
a b s t r a c t
This study demonstrated a one-step process for direct liquefaction and conversion of wet algal biomass
containing about 90% of water to biodiesel under supercritical methanol conditions. This one-step pro-
cess enables simultaneous extraction and transesterication of wet algal biomass. The process conditions
are milder than those required for pyrolysis and prevent the formation of by-products. In the proposed
process, fatty acid methyl esters (FAMEs) can be produced from polar phospholipids, free fatty acids,
and triglycerides. A response surface methodology (RSM) was used to analyze the inuence of the three
process variables, namely, the wet algae to methanol (wt./vol.) ratio, the reaction temperature, and the
reaction time, on the FAMEs conversion. Algal biodiesel samples were analyzed by ATR-FTIR and GC
MS. Based on the experimental analysis and RSM study, optimal conditions for this process are reported
as: wet algae to methanol (wt./vol.) ratio of around 1:9, reaction temperature and time of about 255 C,
and 25 min respectively. This single-step process can potentially be an energy efcient and economical
route for algal biodiesel production.
2010 Elsevier Ltd. All rights reserved.
1. Introduction
Biodiesel can be produced from algal biomass and oils by
extraction-transesterication, direct methanolysis and transesteri-
cation methods (Johnson and Wen, 2009; Belarbi et al., 2000).
Traditionally, algal biodiesel is produced from wet algal biomass
in a series of steps including preparation of dry algae powder,
extraction of algal oils with chemical solvents, and conversion of
the algal oil to biodiesel with a catalyst (Chisti, 2007). Drying the
biomass and extraction of algal oils by conventional methods are
both energy- and cost-intensive. An alternative to the conventional
extraction and transesterication methods is supercritical process.
Using water in wet algae as a tunable co-solvent in supercritical
methanol process not only accelerates the conversion of fats and
algal oils to fatty acid methyl esters (FAMEs), but also increases
solubility and acidity.
In this work, a single-step supercritical process for simulta-
neous extraction and transesterication of wet algal biomass is
demonstrated. In the proposed process, FAMEs can be produced
from polar phospholipids, free fatty acids, and triglycerides by
increasing uidity and volatility while reducing the polarity of
the high-energy molecules in algae at supercritical conditions.
Other advantages with the supercritical process are that they oper-
ate at modest temperatures and have lower energy requirements
compared to conventional extraction and transesterication meth-
ods due to single pot conversion of algal biomass to biodiesel. The
cost of biodiesel production from vegetable oils through supercrit-
ical process is estimated to be $0.26/gal which is about half of that
of the conventional transesterication methods, $0.51/gal (Anite-
scu et al., 2008). This may very well be applied to algal oils as
the algal biomass contains much higher levels of unsaturated fatty
acids, lipids and triglycerides. The objective of the present study is
to demonstrate the direct liquefaction and conversion of wet algal
biomass into biodiesel via a single-step supercritical methanol pro-
cess in the presence of nitrogen, and to optimize the process
parameters that inuence the super critical transesterication
reaction using response surface methodology (RSM).
2. Experimental
2.1. Materials and methods
Algal paste from outdoor open raceways (Inoculum Nannochlor-
opsis sp. (CCMP1776) provided by CEHMM Artesia, NM was used in
0960-8524/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2010.06.031
* Corresponding author. Tel.: +1 575 646 4346; fax: +1 575 646 7706.
E-mail address: sdeng@nmsu.edu (S. Deng).
1
Present address: Solix Biofuels, 430-B North College Ave., Fort Collins, Co 80524,
USA.
Bioresource Technology 102 (2011) 118122
Contents lists available at ScienceDirect
Bioresource Technology
j our nal homepage: www. el sevi er . com/ l ocat e/ bi or t ech
this study. For GCMS analysis, methyl heptadecanoate (C17),
standard for GC, was purchased from Fulka, Milwaukee, WI. Extra
pure (99%) methanol, hexane, acetic acid and sulfuric acid were
purchased from Acros Organics, New Jersey. For the purication
of crude algae FAME, SPE Silica columns were procured from Ther-
mo Scientic, Waltham, MA. The supercritical methanol process
was carried out in the PARR 4593 Micro-reactor with a 4843-con-
troller (Parr Instrument Company, Illinois, USA). Transmission
electron microscopy (TEM) of frozen and residue (after SCM) algal
biomass was examined with a model H-7650 electron microscope
(Hitachi High-Technologies, Pleasanton, CA) operated in the bright
eld mode.
2.2. Characteristics of Nannochloropsis algal species
The ash-free dry weight of the algae sample and lipid yield on
dry weight basis were found to be 69.8% and 50%, respectively. Li-
pid extraction report for Nannochloropsis sp. has the following
composition: triglycerides: 37.74%; other non-polar hydrocarbons,
isoprenoids: 8.72% and polars, glycolipids, phospholipids: 3.54%.
The gross estimation of non-polar hydrocarbons and triglycerides
was done using thin layer chromatography (TLC) and densitometry
technique. Previous work by other researchers (Damiani et al.,
2010; Pyle et al., 2008) showed that total fatty acids accounted
for 3050% of dry biomass, depending on different culture condi-
tions. Qualitative elemental analysis of crude algal biomass was
determined by scanning electron microscopy (SEM, HITACHI S-
3400 N) equipped with energy-dispersive X-ray spectroscopy
(EDS). The major elements and their approximate composition
(wt.%) were carbon (72%), oxygen (21%), sodium (1.5%), magne-
sium (0.41%), silicon (0.93%), phosphorous (0.47%), chlorine
(1.52%), potassium (0.96%).
The FTIR spectra of the Nannochloropsis algal species show the
general features indicating: (i) the highly aliphatic character of
the residues revealed by the strong absorption at 720 cm
1
, (ii)
the presence of hydroxyl groups characterized by the absorption
centered at 3400 cm
1
, and (iii) the presence of carboxyl groups
characterized by the absorption band at 1710 cm
1
, (iv) The pres-
ence of carbonyl groups indicated by the absorption band at about
1735 cm
1
.
2.3. Biosynthesis of triglycerides in microalgae and SCM
transesterication mechanism
The biosynthesis route of triglycerides in microalgae may con-
sist of the following three steps: (a) the formation of acetyl coen-
zyme A (acetyl-coA) in the cytoplasm; (b) the elongation and
desaturation of the carbon chain of fatty acids; and (c) the biosyn-
thesis of triglycerides (Huang et al., 2010). Similar to other higher
plants and animals, microalgae are able to biosynthesize triglycer-
ides to store biomass and energy. In general, L-a-phosphoglycerol
and acetyl-coA are two major elements required for the biosynthe-
sis of triglycerides.
In the supercritical state, depending on pressure and tempera-
ture, the intermolecular hydrogen bonding in the methanol mole-
cule will be signicantly decreased. As a result, the polarity and
dielectric constant of methanol are reduced allowing it to act as
a free monomer. Subsequently, methanol at supercritical condi-
tions can solvate the non-polar triglycerides to form a single phase
of lipid/methanol mixture and yield fatty acid methyl esters and
diglycerides (Saka and Kusdiana, 2001; Kasim et al., 2009). In a
similar way, diglyceride is transesteried to form methyl ester
and monoglyceride, which is converted further to methyl ester
and glycerol in the last step.
2.4. Experimental design
The key variables in the proposed process affecting the FAME
content of the product are the wet algae to methanol (wt./vol.) ra-
tio, the reaction temperature, and the reaction time. A response
surface methodology (RSM) was used (Myers and Montgomery,
2002) to analyze the inuence of these three process variables on
Table 1
Experimental design based on RSM for direct transesterication of wet algal biomass.
Run order Std order Temperature (
o
C) Methanol (wt./vol.) Reaction time (min) Observed FAME% Predicted FAME%
1 1 240 4 10 25.12 24.38
5 2 20.15
6 3 240 4 30 68.13 56.02
26 4 53.15
11 5 240 8 20 38.35 49.32
20 6 240 12 10 32.05 27.37
25 7 28.15
24 8 240 12 30 55.15 59.01
13 9 62.62
16 10 250 4 20 55.56 63.03
3 11 250 8 10 76.05 66.70
12 12 250 8 20 82.15 77.79
22 13 77.20
19 14 83.25
7 15 79.15
14 16 78.15
9 17 76.07
15 18 250 8 30 84.15 88.88
17 19 250 12 20 70.91 77.30
27 20 260 4 10 45.07 44.43
8 21 39.31
18 22 260 4 30 66.37 57.15
21 23 54.12
4 24 260 8 20 68.30 71.19
23 25 260 12 10 70.00 69.97
10 26 72.35
2 27 260 12 30 78.40 82.69
28 28 85.75
P.D. Patil et al. / Bioresource Technology 102 (2011) 118122 119
the fatty acid methyl esters (FAMEs) content. Based on experience
and economic feasibility, a three factorial subset design proposed
by Gilmour (2006) was employed. The design contains three levels
on three factors that could be represented by a cube with six rep-
lications at the center. The six replications at the center offer better
approximation of the true error which statistically helps in deter-
mining signicance of the variables. Another advantage of this
method is its symmetry in design with regard to the center, which
offers equal importance to all levels of all parameters. The total
number of experimental runs was 28 with replications as shown
in Table 1. The wet algae to methanol ratios (wt./vol.), reaction
times, and reaction temperatures were varied in the ranges of
1:41:12, 1030 min, and 240260 C, respectively. The lower
temperature limit, 240 C, was just above the critical temperature
of methanol and the upper temperature limit, 260 C, was deter-
mined by the decomposition temperature limit of algal biomass
(based on several trial runs).
A general second order linear model with the deconstructionist
approach was employed for its exibility and ease of parametric
evaluation for the predicted response surface. Statistically insignif-
icant terms were excluded from the proposed design based on de-
sign hierarchy for the construction of the response surface. Also,
the interaction terms considered manifests a better estimation
on the combination effect of any two parameters considered. Lin-
ear least square method was used to predict the values of param-
eters involved. The condence level of the statistical analysis
conducted was 95%.
2.5. Experimental procedure
The experimental protocol for one-step supercritical methanol
process is as follows: From the aliquots prepared previously (fro-
zen biomass in 50 mL falcon tubes at 80 C), 4 g of wet algae
paste (10% solids) was subjected to a non-catalytic supercritical
methanol (SCM) process in a 100 mL PARR micro-reactor under a
matrix of conditions: constant pressure of 1200 psi; reaction times
of 10, 20, and 30 min; reaction temperatures of 240, 250, and
260 C; and wet algae to methanol (wt./vol.) ratios of 1:4, 1:8,
and 1:12. After the reaction was completed, the reactor contents
were transferred into a 50 mL round-bottom ask and freed of
methanol and volatiles at a reduced pressure in a rotary evapora-
tor. The remaining products were taken in hexane and then centri-
fuged (3200 rpm) for 5 min. The upper organic layer containing
non-polar lipids was extracted and run through a short column
of silica (Hyper SPE Silica). Neutral components were eluted with
the solvent. For qualitative analysis, an internal standard, methyl
heptadecanoate (C17:0) was added quantitatively to the eluted
neutral component-solvent solution and analyzed by gas chroma-
tography-mass spectroscopy (GCMS). The content of the fatty acid
methyl ester in the nal product was calculated quantitatively by
comparing the peak areas of fatty acid methyl esters to the peak
area of the internal standard (methyl heptadecanoate, C17:0) ob-
tained from GCMS.
2.6. Statistical analysis
A general linear model which accounts for the single parame-
ters linear and quadratic effects with their interaction effects
was considered. The following is the general linear model for our
analysis:
l b
0

X
3
i1
b
i
x
i

X
3
i1
b
ii
x
2
i

X
2
i1
X
3
ji1
b
ij
x
i
x
j
where, x
1
, x
2
and x
3
are the levels of the factors and l is the pre-
dicted response if the process were to follow the model. A decon-
structionist approach was followed which indicates the
consideration of a complete quadratic model and eliminating terms
which were not signicant as the analysis continued. All further
analysis was carried out using both coded and uncoded variables.
Method of least squares was employed to ascertain the values of
the model parameters and ANOVA to establish their statistical sig-
nicance at a condence level of 95% (in our case).
3. Results and discussion
3.1. Development of regression model
The central composite design (CCD) matrix and the response
obtained from the experimental runs are shown in Table 1. The
estimated regression coefcients for response and analysis of var-
iance (ANOVA) for response are shown in Tables 2.1 and 2.2,
respectively.
The regression analysis indicates that all the three parameters
had signicant inuence on the fatty acid methyl ester content,
which is conrmed by the P-values. The response surfaces were t-
ted using process variables that were found to be signicant after
the analysis. The P-value of the lack of t analysis is 0.133, which
is more than the 0.05 (condence level is 95%). The regression
model provides accurate description of the experimental data indi-
cating successful correlation among the three transesterication
process parameters that affect the yield of algal biodiesel. This is
further supported by the correlation coefcient, R
2
of 0.921.
3.2. Effects of process parameters and optimization
Fig. 1shows the response contours of FAMEyieldagainst reaction
temperature and wet algae to methanol (wt./vol.) ratio at three dif-
ferent reactiontimeintervals andxedreactionpressureof 1200psi.
The values and signs on the regression coefcients suggest that the
reaction time affects the response positively for temperatures up
to 255 C; however, reaction temperatures above 255 C were not
suitable for transesterication reaction of the algal biomass at xed
Table 2.1
Estimated regression coefcients for response (the analysis was done using coded
units).
Term Coef SE Coef T P Signicant
Constant 77.012 1.938 39.73 0 Yes
Temp 10.933 1.337 8.179 0 Yes
Meth 7.133 1.337 5.336 0 Yes
Time 11.088 1.337 8.295 0 Yes
Temp Temp 20.213 3.38 5.981 0 Yes
Meth Meth 10.303 3.38 3.048 0.007 Yes
Time Time 6.562 3.38 1.942 0.068 No
Temp Meth 5.638 1.418 3.977 0.001 Yes
Temp Time 4.729 1.418 3.336 0.004 Yes
Meth Time 2.047 1.418 1.444 0.166 No
R
2
= 94.46%, R-Sq (pred) = 84.35%, R-Sq (adj) = 91.69%.
Table 2.2
Analysis of variance for response.
Source DF Seq SS Adj SS Adj MS F P
Regression 9 9870.7 9870.7 1096.74 34.1 0
Linear 3 5280.7 5280.7 1760.23 54.73 0
Square 3 3656.5 3656.5 1218.83 37.9 0
Interaction 3 933.5 933.5 311.17 9.67 0.001
Residual error 18 578.9 578.9 32.16
Lack-of-t 5 257.6 257.6 51.51 2.08 0.133
Pure error 13 321.4 321.4 24.72
Total 27 10449.6
120 P.D. Patil et al. / Bioresource Technology 102 (2011) 118122
pressure of 1200psi. This may be due to the fact that the oil/lipidand
the alkyl esters tend to decompose or become thermally unstable
above the specied temperature owing to the high content of unsat-
urated fatty acids (Gui et al., 2009). It was observedthat at hightem-
perature and pressure, unsaturated fatty acids tend to decompose
due to the isomerization of the double bond functional group from
cis-type carbon bonding (C@C) into trans-type carbon bonding
(C@C), which are naturally unstable fatty acids (Imahara et al.,
2008). The FT-IR spectrum (reaction temperature of 270 C) shows
a trans-type carbon bonding (C@C) group at the wavelength of
960 cm
1
which is absent at this wavelength for the reaction tem-
perature of 250 C. The greater the percentage of unsaturation in
fattyacids or esters of algal biomass, the more it is susceptible tooxi-
dation. The extent of unsaturation of in esters can be reduced easily
by partial catalytic hydrogenation of the oil (Dijkstra, 2006).
Wet algae to methanol (wt./vol.) ratios have a positive effect on
the yield up to 1:9 but have a negative impact at higher levels.
Higher ratio of biomass to methanol could shift the reversible reac-
tion forward (as observed) perhaps due to increased contact area
between methanol and lipid, resulting in higher yield of FAME
and it also contributes to the lower critical temperature of the mix-
ture. However, its interaction with reaction temperature can, on
the other hand, cause a reduction in the yield of FAME due to either
the decomposition of FAME or the critical temperature of the reac-
tant/product mixture between methanol and FAMEs becomes
highly dependent on the concentration of methanol and may de-
crease the critical temperature of the reactant/product. When reac-
tant/product mixture is heated above critical temperature, it has
the tendency to decompose (Hegel et al., 2008).
As expected, a longer reaction time allows the transesterica-
tion reaction to proceed to completion and results in a higher yield
of FAMEs from algal biomass. According to Vieitez et al. (2009),
higher reaction time beyond particular limit in supercritical alco-
hol process for vegetable oil may lead to greater losses of unsatu-
rated FAME due to degradation reactions. Nevertheless, Fig. 1
shows that the effect of reaction time is more prominent at
wet algae to methanol (wt./vol.) ratio of 1:9 and reaction temper-
ature around 255 C at a xed reaction pressure of 1200 psi. In
water added supercritical methanol reaction, the watermethanol
mixture has both strong hydrophilic and hydrophobic properties
that help speeding up the reaction signicantly (Kusdiana and
Saka, 2004). Based on the experimental analysis and RSM study,
the optimal conditions for this process are reported as:
wet algae/methanol (wt./vol.) ratio of around 1:9, reaction temper-
ature and time of about 255 C, and 25 min respectively.
3.3. TEM of frozen algal biomass (raw material) and residual
Nannochloropsis algal sample after SCM process
For analysis of elemental composition, raw and residual sam-
ples were washed with distilled water and centrifuged pellets were
excised from centrifuge tubes, air-dried and glued to carbon adhe-
sive tabs (Electron Microscopy Sciences, Hateld, PA) on aluminum
sample stubs. Elemental spectra were collected at 15 kV using a
model S-3400 N scanning electron microscope (Hitachi High-Tech-
nologies, Pleasanton, CA) equipped with a model Noran System Six
300 energy-dispersive spectrometer system (Thermo Electron
Corp., Madison, WI).
From TEM analysis report of frozen (raw) and residual algal bio-
mass, it was found that at SCM condition, algal cell wall structure
was totally disturbed and fragmented while EDS report showed the
evidence for thermal degradation of algal biomass (wt.% of C in-
creased in residue) due to high content of unsaturated fatty acids
in lipid.
3.4. Analysis of algal biodiesel conversion
For the quantication of reaction product, the algal biodiesel
samples were analyzed by a gas chromatographymass spectrom-
etry (GCMS) system incorporated with an Agilent 5975 C MSD
and an Agilent 7890 A GC. The content of the fatty acid methyl es-
ter in the nal product was calculated quantitatively by comparing
the peak areas of fatty acid methyl esters to the peak area of the
internal standard (methyl heptadecanoate, C17:0) obtained from
GCMS. It is noted from GCMS results that algal biodiesel con-
tains a major proportion of mono and poly unsaturated fatty acid
methyl esters. The major fatty acids were palmitoleic acid
(C16:1, 3033%), oleic acid (C18:1, 3538%), eicosapentanoic acid
(EPA, C20:5n3, 58%), palmitic acid (C16:0, 510%,) and arachi-
donic acid (C20:4n6, 13%). From the GCMS peak and total ion
chromatography (TIC) data, it was observed that the algal biodiesel
contains olens, fatty alcohols and sterols in minor quantities
along with saturated and unsaturated FAMEs. The relative weight
compositions of organic compounds present in the algal biodiesel
were analyzed using GCMS, and summarized in Table 3.
ATR-FTIR spectra of petro-diesel, camelina biodiesel, and algal
biodiesel are shown in Fig. 2. The IR spectra were obtained using
a PerkinElmer Spectrum 400 FT-IR/FT-NIR spectrometer. The main
components of diesel are aliphatic hydrocarbons, whose chemical
structures are similar to long carbon chain of the main components
of biodiesel. The observation of an absorption Peaks around
1200 cm
1
may be assigned to the antisymmetric axial stretching
vibrations of CC(@O)O bonds of the ester, while peaks around
1183 cm
1
may be assigned to asymmetric axial stretching vibra-
tions of OCC bonds (Silverstein and Webster, 1998). In addition,
Fig. 1. FAME yield against reaction temperature and wet algae wt/methanol
volume ratio at different reaction times using RSM.
P.D. Patil et al. / Bioresource Technology 102 (2011) 118122 121
since biodiesel is mainly mono-alkyl ester, the intense C@O
stretching band of methyl ester appears at 1743 cm
1
for algal
and camelina biodiesel which is absent in petro-diesel spectra.
4. Conclusions
The single-step process for wet algal biomass (Inoculum Nanno-
chloropsis sp., CCMP1776) may offer the benets of shorter reaction
time, simple purication of products and maximum conversion of
triglycerides into their corresponding fatty acid methyl esters. The
single-step process favors the energy requirements for biodiesel
production by eliminating the needs for drying and extraction of
algal biomass. Process optimization using response surface meth-
odology (RSM) design proved to be a valuable tool for evaluating
the effects of the process variables on the FAMEs yield. The sin-
gle-step process has the potential to provide an energy efcient
and economical route to algal biodiesel production.
Acknowledgements
This project was partially supported by New Mexico State Uni-
versity Ofce of Vice President for Research and State of New Mex-
ico through a New Mexico Technology Research Collaborative
grant. The authors are thankful to CEHMM Artesia, NM for provid-
ing the wet algal biomass for biodiesel testing.
References
Anitescu, G., Deshpande, A., Tavlarides, L., 2008. Integrated technology for
supercritical biodiesel production and power cogeneration. Energy Fuels 22,
13911399.
Belarbi, E.-H., Molina, G.E., Chisti, Y., 2000. A process for high yield and scalable
recovery of high purity eicosapentaenoic acid esters from microalgae and sh
oil. Enzyme Microb. Technol. 26, 516529.
Chisti, Y., 2007. Biodiesel from microalgae. Biotechnol. Adv. 25, 294306.
Damiani, C., Popovich, C., Constenla, D., Leonardi, P., 2010. Lipid analysis in
Haematococcus pluvialis to assess its potential use as a biodiesel feedstock.
Bioresour. Technol. 101, 38013807.
Dijkstra, A.J., 2006. Revisiting the formation of trans isomers during partial
hydrogenation of triacylglycerol oils. Eur. J. Lipid Sci. Technol. 108, 4964.
Gilmour, S.G., 2006. Response surface designs for experiments in bioprocessing.
Biometrics 62, 323331.
Gui, M., Lee, K., Bhatia, S., 2009. Supercritical ethanol technology for the production
of biodiesel: process optimization studies. J. Superc. Fluids 49, 286292.
Hegel, P., Andreatta, A., Pereda, S., Bottini, S., Brignole, E., 2008. High pressure phase
equilibria of supercritical alcohols with triglycerides, fatty esters and co-
solvents. Fluid Phase Equilib. 266, 3137.
Huang, G., Chen, F., Wei, D., Zhang, X., Chen, G., 2010. Biodiesel production by
microalgal biotechnology. Appl. Energy 87, 3846.
Imahara, H., Minami, E., Hari, S., Saka, S., 2008. Thermal stability of biodiesel in
supercritical methanol. Fuel 87, 16.
Johnson, M., Wen, Z., 2009. Production of biodiesel fuel from the microalga
Chizochytrium limacinum by direct transesterication of algal biomass. Energy
fuels 23, 51795183.
Kasim, N., Tsai, T.-H., Gunawan, S., Ju, Y.-H., 2009. Biodiesel production from rice
bran oil and supercritical methanol. Bioresour. Technol. 100, 23992403.
Kusdiana, D., Saka, S., 2004. Effects of water on biodiesel fuel production by
supercritical methanol treatment. Bioresour. Technol. 91, 289295.
Myers, R.H., Montgomery, D.C., 2002. Response Surface Methodology: Process and
Product Optimization Using Designed Experiments. Response Surface
Methodology, second ed. Wiley, New York.
Pyle, D.J., Garcia, R.A., Wen, Z.Y., 2008. Producing docosahexaenoic acid (DHA)-rich
algae from biodiesel-derived crude glycerol: effects of impurities on DHA
production and algal biomass composition. J. Agric. Food Chem. 56, 39333939.
Saka, S., Kusdiana, D., 2001. Biodiesel fuel from rapeseed oil as prepared in
supercritical methanol. Fuel 80, 225231.
Silverstein, R., Webster, F., 1998. Spectrometric identication of organic
compounds, sixth ed. Wiley, New York.
Vieitez, I., Silva, C., Alckmin, I., Borges, G.R., Corazza, F., Oliveira, J.V., Grompone,
M.A., Jachmanin, I., 2009. Effect of temperature on the continuous synthesis of
soybean esters under supercritical ethanol. Energy Fuels 23, 558563.
Table 3
GCMS peak TIC data of crude biodiesel obtained from algal biomass.
Peak Retention time Area (%) Name
1 3.264 0.63 2H-pyran
2 3.802 0.03 Octanoic acid methyl ester
3 4.609 0.01 Nonanoic acid methyl ester
4 4.905 0.2 Dodecane
5 5.250 0.1 Benzenamine
6 5.547 0.1 Indole
7 5.759 0.35 Decanoic acid methyl ester
8 6.354 0.18 Naphthalene,2,6-dimethyl
9 7.933 0.05 cis-5-Dodecenoic acid methyl ester
10 8.305 3.14 Undecanoic acid, 10-methyl ester
11 9.524 19.72 Tridecanoic acid methyl ester
12 10.245 0.02 8-Heptadecene
13 13.587 1.74 7-Hexadecenoic acid methyl ester
14 13.632 4.5 Cyclopropaneoctanal,2-octyl-9-Eicosyne
15 14.125 0.2 Cyclohexaneethanol
16 15.124 7.34 9-Hexadecenoic acid methyl ester
17 16.250 10.2 Tricyclo decane
18 16.923 9.92 Pentadecanoic acid,13-methyl ester
19 17.089 33.28 9-Octadecenoic acid methyl ester
20 17.878 8.29 Eicosanoic acid methyl ester
Fig. 2. FTIR results of algal biodiesel, petro-diesel, and camelina biodiesel.
122 P.D. Patil et al. / Bioresource Technology 102 (2011) 118122