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Procedia Engineering 42 (2012) 231 – 238

20th International Congress of Chemical and Process Engineering CHISA 2012

25 – 29 August 2012, Prague, Czech Republic

Biofuels from algae

A. Kleinováa a*, Z. Cvengrošováa, J. Rimarčíka, E. Buzetzkia, J. Mikulecb,
J. Cvengroša
a
Faculty of Chemical and Food Technology STU, Radlinského 9, 812 37 Bratislava, Slovakia
b
Slovnaft VÚRUP, a.s., Vlčie hrdlo, 820 03 Bratislava, Slovakia

Abstract

Fatty acid methyl esters (FAME) are used as alternative diesel fuel originating from renewable sources.
The attention is focused on the materials that do not compete with food and feed production. Algae have a
significant potential as an alternative biodiesel feedstock. In comparison to other crops, they have the
advantage in very fast reproduction cycles, enhanced resistance to high UV radiation doses and higher
effectiveness of energy conversion to biomass due to low demands on other metabolic functions.
Moreover, they fix waste CO2 very efficiently. The key problem is to destroy the algae cell in order to
obtain the oil. In this work, the results of FAME preparation from received algae oils are presented. This
study confirms that Nannochloropsis and Chlorella microalgae provide valuable triacyglycerols for the
biofuel production. Characteristics of prepared FAME meet the EN 14 214 standard requirements.
However, FAME from algae oil may exhibit impaired oxidative stability due to higher level of
unsaturation.

© 2012 Published by Elsevier Ltd. Selection under responsibility of the Congress Scientific Committee
(Petr Kluson) Open access under CC BY-NC-ND license.

Keywords: Alternative fuels; FAME; algae

* Corresponding author. Tel.: 00421259325538; fax: 00421259325751.

E-mail address: andrea.kleinova@stuba.sk

1877-7058 © 2012 Published by Elsevier Ltd. Open access under CC BY-NC-ND license.
doi:10.1016/j.proeng.2012.07.414
232 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238

Nomenclature

AV Acid value
BHT Buthylhydroxytoluene
FAME Fatty Acid Methyl Esters
TAG Triacylglycerols

1. Introduction

The demand for liquid fuels in transport is rising. Nowadays, fatty acid methyl esters (FAME) are
accepted liquid biofuels for diesel engines. They are usually prepared from vegetable oils or animal fats.
As these articles are dedicated mainly for consumption, other renewable sources of natural
triacylglycerols (TAG) are sought.
The idea of microalgae utilization in the fuel production is not new [1]. Using microalgae in the
biofuel production will not compete with the production of food, fodder and other products from crops.
However, not every algal species are satisfactory for biofuel producing.
Algae are recognized as one of the oldest life-forms and also as the worldwide fastest growth plants.
These phototrophic organisms require for optimal growth sunlight, CO 2 from the air, water, inorganic
salts (N, P, K) and temperature in the range of 2030 °C. Microalgae can fix CO2 from three different
sources: atmosphere, discharge gases from heavy industry and from soluble carbonates [2]. Microalgal
biomass contains approximately 50 % of carbon by dry weight [3]. Producing 100 t of algal biomass fixes
roughly 183 t of CO2.
Depending of species, microalgae produce many different kinds of lipids, hydrocarbons and other
complex oils [46]. In general, many algae species have the oil content ranging from 20 to 50 % by dry
weight of biomass. The lipid and fatty acid contents of microalgae vary in accordance with culture
conditions. It is possible to increase the lipid concentration by optimizing the growth determining factors
[7] almost up to 80 %. Oil productivity, the mass of oil produced per unit volume of the microalgal broth
per day, depends on the algal growth rate and the oil content of the biomass. The yield of the oil produced
by algae is significantly higher (100000 L/ha) in comparison to other crops, for example soybean (446
L/ha), sunflower (952 L/ha), rapeseed (1200 L/ha), castor (1413 L/ha), coconut (2689 L/ha) and palm
(5950 L/ha).
There exist three distinct algae production mechanisms including photoautotrophic, heterotrophic and
mixotrophic production. Currently, photoautotrophic production is the only method which is technically
and economically feasible for large-scale production of algae biomass for non-energy production [8].
Two developed systems are based on open pond and closed pond photobioreactor technologies [9]. High
algae production rates are achievable with open pond systems.
The recovery of microalgal biomass which generally requires one or more solid-liquid separation steps
is a challenging phase of the algal biomass process [2], and accounts for 2030 % of the total costs of
production [10]. The selection of harvesting technology is crucial to economic production of algal
biomass [11]. The processes involved include flocculation, filtration, flotation, centrifugal sedimentation,
also extraction and purification. Well-known methods of the oil extraction are: (i) Expeller/Press, (ii)
solvent extraction and (iii) supercritical fluid extraction. The most popular chemical solvent for extraction
is hexane. More efficient that solvent separation is supercritical extraction. Supercritical fluids are
 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238 233

selective, thus providing the high purity and product concentration. This can extract almost 100 % of the
oils all by itself.
The Turkey Soley Institute Company has developed high sophisticated and high efficient processing
technology of algal biomass with the oil daily production capacity of 320 m3. Company is able to produce
1.7 kg microalgae in a day in 1 m3 water media [12]. Used oily microalgae have almost 50 % of oil
content. It means that the company produces 600 L microalgae oil in 1m2 area in a year by a high-tech
photobioreactor, while the production of rapeseed oil is only 0.125 L in 1m2 area in a year. They can
place 500.000 L water capacity photobioreactor in 112m2 area. The oil production involves microalgae
growth, bacterial cracking, ultrasonic extraction, microwave extraction, solvent extraction (water), cold
pressing, filtration and UV sterilization. The utilization of the by-product is also reasonably solved.
Figure 1 shows the adapted scheme of the oil extract ion in the company.

Extractor
bacteria

Microalgae sludge
Microalgae (contains almost
can be duplicated 50% of oil)
3-4 times in a day 4 hours 2 hours

Microalgae production in photobioreactor Bacterial cracking Ultrasonic Extraction Microwave Extraction

systems by organic fertilizers Microalgae harvest (Extractor bacteria can
filters can collect crack microlagae cell walls )
almost 1.7 g microalgae 2 hours
from each liter of water
media in a day

Filtered water media - back to photobioreactor

Solvent Extraction
(By Water)
Microalgal Dregs
2 hours
Microalgal Dregs
for fertilizer and Oil Water Mixture
animal feed Cold Press
as final by-product

Separated Water
Oil - Water Separation
Oil Filter and UV Sterilization

Microalgae Oil
as final product

Fig. 1. Scheme of algae oil extraction process in Soley Institute, Turkey (adapted from [12])

The conversion of algal biomass-to-energy encompasses different processes ordinarily used for
terrestrial biomass and which depend, to a large extent, on the types and sources of biomass, conservation
234 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238

options and end-use [13]. The conversion technologies for microalgae utilization can be divided into two
basic categories of thermochemical (gasification, thermochemical liquification, pyrolysis, direct
combustion) and biochemical conversion (anaerobic digestion, alcoholic fermentation, photobiological
hydrogen production). Potential products beside electricity production are syngas, bio-oil, charcoal,
methane, hydrogen, ethanol, hydrogen [14].
Due to many advantages and developed technologies of algae cultivation and processing, algae seem
to be promising commodity for the primary source of biofuels. However, high processing costs and low
competitiveness to fossil diesel are the major obstacles in their extensive commercial utilization.
Presented research work deals with the qualitative assessment of obtained algae oil samples and
prepared methyl esters as alternative liquid fuels for diesel engines according to EN 14 214 standard.

2. Materials and Methods

2.1 Materials

Refined rapeseed oil Clever, Germany, was used as a standard. Three received algae oil samples were
studied. The algae oil sample for cosmetic purposes (Algae1) was purchased from Sprinnrad, Germany.
This sample contains TAG from Focus Vesiculosus algae. The second algae oil (Algea2) was purchased
from Creativ Cosmetik, Salzburg, Austria, and was of Nannochloropsis microalgae origin. The last
Chlorella microalgae oil sample (Algae3) was received from Soley Institute, Turkey.
The acidity, density, viscosity and acyl profile were determined in each oil sample. The results are
given in Tables 1 and 2.
To determine the acidity, following chemicals were used: KOH in ethanol and ethanol:toluene 1:1
(v/v) as a solvent. Transesterification was performed with 5.3 wt. % KOH in methanol as homogeneous
catalysts. For oxidative stability measurements, prepared FAME samples were stabilized using the
buthylhydroxytoluene (BHT) as the antioxidant in the amount of 0.1 wt. %.

2.2 Methods

FAME were prepared by two-step alkali catalyzed transesterification of TAG with methanol [15].
Unreacted methanol was removed from the mixture on rotary evaporator RVO 64 at 80 °C and reduced
pressure (ca 1000 Pa). Residual glycerol was removed by means of centrifugation (ca 1500 g). FAME
were than washed by adding of 1 wt. % of water, centrifuged in order to remove water and filtered.
Conversion degree of acylglycerols to FAME was determined by the method referred to in [16]. The
method is based on the comparison of FAME peak areas in the sample prior to and after the sample
treatment with an efficient transesterification agent. Analyses were measured using the gas
chromatograph HP 5890 with flame ionization detector and 1.8 m glass packed column of internal
diameter of 0.3 cm. 10 % SE 30 on Chromatone NAW DMCS, particle size of 0.125 – 0.16 mm was used
as the stationary phase. Samples were analyzed isothermally at 250 °C. The injector temperature was 240
°C, the detector temperature 270 °C. Nitrogen with flow rate of 30 cm3 min-1 was used as a carrier gas.
Viscosities of oil and FAME samples were determined according STN ISO 3104 standard using
Ubbelohde viscomer with 0.0998 calibration constant at the temperatures of 40, 50, 60 a 70 °C. The final
value represents the average of three measured viscosity values.
The densities were determined using 50 ml pycnometer at room temperature. The final value
represents the average of two measured densities and was recalculated to 15 °C as follows:

U15°C) = U(t) + k·(t15) (1)

 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238 235

where the k was 0.723 in the case of FAME. On the other hand, k of 0.650 was used in the case of oils.
The acid values (AV) of studied samples were determined by titration according to STN 58 8756.
Another qualitative parameter measurements for complete qualitative assessment of FAME according
to EN 14 214 were carried out in external laboratories. These measurements comprised standard methods.

3. Results and discussions

Table 1 demonstrates determined acyl portions in the obtained algae oil samples. Based on
measurements results, the Algae1 oil sample was rejected from the study. This sample is unsuitable for
FAME preparation due to the presence of C8 and C10 acyls. The method of ester content determination
(EN 14 103) monitors the acyls ranging from C12 to C 36. Therefore, this sample would give zero value
of ester content. The ester content is among important parameters of FAME qualitative assessment
according to EN 14 214. The oil from Nannochloropsis microalgae (Algae2) contains predominately
acyls of C18:2, C18:1, while C16:0 is in small amount. Its composition is similar to sunflower oil. The oil
produced by Chlorella microalgae (Algae3) consists mainly of C18:1, C18:2 and also of C18:3 and C16:0
in small amounts. The acyl profile is comparable to high oleic sunflower oil.

Table 1. Acyl profiles determined by gas chromatography

C8:0 C10:0 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 6other
Sample
(%) (%) (%) (%) (%) (%) (%) (%) (%) (%)
Algae1 oil 55 42.6        2.4
Algae2 oil    6.7 0.2 3.3 33.9 54.5 0.4 0.65
Algae3 oil   0.07 5.03 0.3 1.33 63.2 21.4 7.1 1.57

Obtained algae oil samples have fundamental physico-chemical properties close to chosen standard –
rapeseed oil (Table 2). These results also confirm the unsuitability of Algae1 oil sample for FAME
preparation. Its viscosity and density values correspond to the presence of caprylic triacylglycerols as a
dominant component.

Table 2. Fundamental physico-chemical characteristics of studied oils

Oil sample AV (mg KOH/g) ν40°C (mm2 s-1) ρ15°C (kg m-3)
Rapeseed oil 0.2 35.546 919.7
Algae1 oil 0.2 14.627 946.3
Algae2 oil 0.2 31.437 920.6
Algae3 oil 0.2 33.433 919.1

On the basis of obtained results from algae oils analyses, Algae2 and Algae3 oil samples were selected
for FAME preparation. Some parameters of prepared FAME are summarized in the Table 3. High
conversion rates have been observed in both samples. The samples also exhibit sufficient ester contents.
Other monitored parameters agree with the EN 14 214 limits with the exception of oxidation stability in
the case of FAME from Algae2. The oxidation stability was not significantly improved even by adding of
0.1 wt. % of antioxidant BHT. Based on gas chromatography results (Table 2), reduced oxidation stability
may be expected because of higher amount of unsaturated fatty acids in used oil sample. In the literature,
we can find information about impaired oxidation stability of algae oils. Some algae oils may contain four
236 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238

or even more double bonds [17]. The extent of unsaturation can be modified by partial hydrogenation
using the same technology as that used in the margarine production [18, 19]. Literature reveals the
relative rate of oxidation for methyl esters of C18:1, C18:2, and C18:3 acids to be 1:12:25 [20]. Another
source refers oxidation rates for oleates of 1, for linoleates of 41 and for linolenates even of 98 [21, 22].

Table 3. Parameters of prepared FAME from studied algae oils

FAME FAME
Parameter Unit Method EN 14 214
Algae2 Algae3

Conversion wt. % [16] – 100 98

Ester content wt. % STN EN 14103 min. 96.5 99.8 97.5

Acid value mg KOH/g STN 58 8756 max. 0.5 0.2 0.3

EN ISO 3675/
Density kg/m3 860–900 883.9 884.8
EN ISO 12185

Viscosity mm2/ s EN ISO 3104 3.55 4.3 4.99

Methanol content wt. % STN EN 14110 max 0.20 0.02 0.018

Monoacylglycerols
wt. % STN EN 14105 max 0.80 0.85 0.58
content

Diacylglycerols
wt. % STN EN 14105 max 0.20 0.22 0.17
content

Triacylglycerols
wt. % STN EN 14105 max 0.20 0.28 0.25
content

Free glycerol
wt. % STN EN 14105 max 0.02 0.006 0.003
content

Glycerol content wt. % STN EN 14105 max 0.25 0.28 0.21

Na content mg/kg STN EN 140108

max 5.0
K content mg/kg STN EN 14109 1.154 2.05

Ca content mg/kg STN EN 145038 n.d. 0.64

max 5.0
Mg content mg/kg STN EN 145038 0.952 1.24

P content mg/kg ASTM D 3231 max 4.0 1.798 2.18

Oxidation stability,
h EN 14112 min 6.0 1.22/4.85* 6.9/12.5*
110 °C
* after adding of BHT
 A. KleinovaÅL et al. / Procedia Engineering 42 (2012) 231 – 238 237

4. Conclusion

FAME preparation from algae oils as potential alternative liquid fuels for diesel engines was studied in
this work. The research activities were focused on physico-chemical characterization of received algae oil
samples, assessment of their suitability for FAME preparation, subsequent preparation, final treatment
and analytical evaluation of FAME in order to comply EN 14 214 requirements. Obtained results showed,
that Nannochloropsis and Chlorella microalgae provided valuable triacylglycerols for the FAME
preparation by two-step alkali catalyzed transesterification. It can be concluded that prepared FAME are
not significantly different that those prepared from vegetable oils and meet the requirements of the EN
14 214 standard. One FAME sample exhibited impaired oxidative stability, less than 6 h, due to the
presence of relatively high amount of acyls with two or three double bonds. Used antioxidant BHT in the
concentration of 0.1 wt. % had no significant effect on oxidative stability of studied FAME sample.

Acknowledgements

This work was supported by the Slovak Research and Development Agency under the contract No.
APVV-0665-10.

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