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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. Kleinova a*, Z. Cvengroova, J. Rimarka, E. Buzetzkia, J. Mikulecb, J. Cvengroa
a

Faculty of Chemical and Food Technology STU, Radlinskho 9, 812 37 Bratislava, Slovakia b Slovnaft VRUP, a.s., Vlie 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)
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. doi:10.1016/j.proeng.2012.07.414

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Nomenclature AV BHT TAG Acid value Buthylhydroxytoluene Triacylglycerols

FAME Fatty Acid Methyl Esters

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 20 30 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 20 30 % 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

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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 can be duplicated 3-4 times in a day

Microalgae sludge (contains almost 50% of oil)

4 hours

2 hours

Microalgae production in photobioreactor systems by organic fertilizers

Microalgae harvest filters can collect almost 1.7 g microalgae from each liter of water media in a day

Ultrasonic Extraction Bacterial cracking (Extractor bacteria can crack microlagae cell walls )

Microwave Extraction 2 hours

Filtered water media - back to photobioreactor

Solvent Extraction (By Water) Microalgal Dregs 2 hours

Microalgal Dregs for fertilizer and animal feed as final by-product

Oil Water Mixture

Cold Press

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

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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:

U15C) = U(t) + k(t15)

(1)

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

Sample Algae1 oil Algae2 oil Algae3 oil

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 Rapeseed oil Algae1 oil Algae2 oil Algae3 oil AV (mg KOH/g) 0.2 0.2 0.2 0.2 40C (mm2 s-1) 35.546 14.627 31.437 33.433 15C (kg m-3) 919.7 946.3 920.6 919.1

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

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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 Algae2 100 99.8 0.2 883.9 4.3 0.02 0.85 FAME Algae3 98 97.5 0.3 884.8 4.99 0.018 0.58

Parameter Conversion Ester content Acid value Density Viscosity Methanol content Monoacylglycerols content Diacylglycerols content Triacylglycerols content Free glycerol content Glycerol content Na content K content Ca content Mg content P content Oxidation stability, 110 C * after adding of BHT

Unit wt. % wt. % mg KOH/g kg/m3 mm2/ s wt. % wt. %

Method [16] STN EN 14103 STN 58 8756 EN ISO 3675/ EN ISO 12185 EN ISO 3104 STN EN 14110 STN EN 14105

EN 14 214 min. 96.5 max. 0.5 860900 3.55 max 0.20 max 0.80

wt. %

STN EN 14105

max 0.20

0.22

0.17

wt. %

STN EN 14105

max 0.20

0.28

0.25

wt. %

STN EN 14105

max 0.02

0.006

0.003

wt. % mg/kg mg/kg mg/kg mg/kg mg/kg h

STN EN 14105 STN EN 140108

max 0.25

0.28

0.21

max 5.0 STN EN 14109 STN EN 145038 max 5.0 STN EN 145038 ASTM D 3231 EN 14112 max 4.0 min 6.0 0.952 1.798 1.22/4.85* 1.24 2.18 6.9/12.5* 1.154 n.d. 2.05 0.64

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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. References [1] [2] [3]
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