J Am Oil Chem Soc DOI 10.

1007/s11746-011-1963-0

ORIGINAL PAPER

Synthesis and Characterization of New Biodiesels Derived From Oils of Plants Growing in Northern Wisconsin and Minnesota
James W. Lane Kurt Hukriede Adam Jersett Damodar Koirala Dan Levings Aaron Stewart Michael A. Waxman

Received: 26 July 2011 / Revised: 12 October 2011 / Accepted: 17 October 2011 AOCS 2011

Abstract We report on the preparation and selected properties of some new biodiesels which we synthesized from oils of plants growing in Northern Wisconsin and Minnesota. The composition and the low-temperature properties such as crystallization onset Tc and end of melting Tm investigated with the help of differential scanning calorimetry are presented. Some of these biodiesels exhibited remarkably good low-temperature characteristics. In order to further improve these properties, we use a variety of alcohols during the transesterication process, including isopropyl, 2-butyl, and isoamyl alcohols. Using several parameters such as oil content and crystallization onset temperature Tc, plant species that appear most promising have been identied, among those highbush cranberry (Tc & -31 C for its methyl esters, Tc & -41 C for its 2-butyl esters), dotted horsemint (Tc & -17 C for its methyl esters, Tc & -40 C for its 2-butyl esters), and American hazelnut (Tc & -19 C for its methyl esters, Tc & -30 C for its 2-butyl esters). Keywords Biodiesel feedstocks Branched-chain esters Differential scanning calorimetry Crystallization

Introduction Modern society gives a high priority to increased usage of renewable resources. In the last two decades, signicant

J. W. Lane (&) K. Hukriede A. Jersett D. Koirala D. Levings A. Stewart M. A. Waxman Chemistry and Physics Program, Department of Natural Sciences, and Lake Superior Research Institute, University of Wisconsin-Superior, Superior, WI 54880, USA e-mail: jlane@uwsuper.edu

efforts have been made in order to identify biodiesels that can replace traditional fossil fuels. Unfortunately, most biodiesels produced from the traditional feedstocks such as soybean oil develop various performance issues once the ambient temperatures approach 0 C, which impedes their usage in cold climates [1]. That is one of the main reasons why a number of studies (e.g. sources [14]) have recently appeared describing biodiesels produced from alternative feedstocks. Biodiesel contains esters of both saturated and unsaturated fatty acids, with the latter ones freezing at considerably lower temperatures than the corresponding saturated esters. As we will demonstrate below, many biodiesels derived from northern plants contain increased amounts of unsaturated esters and thus exhibit improved low-temperature characteristics compared to soybean-derived biodiesel. In addition, many of these feedstocks can be cultivated on non-arable lands (see, for example, references [58]) and thus do not compete with common food crops. Low-temperature properties of these biodiesels can be further improved by using branched alcohols in the production process [1, 9]. Indeed, crystallization involves alignment of molecules into an orderly pattern. As branches are introduced into the molecular structure of esters, such an alignment becomes impeded and crystallization temperature, Tc, is reduced. In the above study [9] where soybean had been used as the feedstock, the value of Tc was diminished by as much as 14 C by producing isopropyl and 2-butyl esters rather than methyl esters; similar results can be expected with other oil sources. The composition of biodiesel, including both the fatty acid content and minor constituents such as tocopherols, tocotrienols, and phytosterols, varies depending upon the feedstock, which in turn inuences chemical and physical properties; thus, mixing biodiesels obtained from different

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feedstocks could affect the biodiesel properties [10]. Similarly, these properties can also be affected by mixing biodiesels obtained from a single feedstock with different alcohols [11]. In the nal part of this study, we briey analyze both kinds of mixtures, again with regard to their low-temperature characteristics.

Fatty Acid Alkyl Ester Preparation A procedure similar to that described by Wang et al. [13] for producing isopropyl esters was followed. In a typical procedure a 20:1 alcohol:oil molar ratio was used. 2-butanol alcohol (16.91 g, 0.228 mol) was added to a dry ask tted with a condenser and drying tube. Sodium metal (0.108 g, 1 wt% based on oil) was added to the alcohol and the mixture was heated to 80 C for approximately 45 min until all the sodium had reacted. Oil (9.808 g) was then added and stirring was continued for 12 h at 85 C. Hexanes (25 mL) was added to the warm solution and the organic layer was extracted four times with 20 mL of water held at 6070 C, and then two times with 10 mL saturated sodium chloride solution. The hexane-biodiesel mixture was dried with anhydrous sodium sulfate and hexanes removed under vacuum (10 mmHg at 50 C for 15 min) until a constant weight was obtained. A sample of the alkyl esters (9.520 g) was puried further by ltering through 2.3 g of silica using 30 mL hexanes followed by 10 mL of 90:10 hexanes:ether. The yield of puried alkyl esters was 9.234 g. The sample showed a single peak corresponding to alkyl esters when analyzed by HPLCELSD. Other alkyl esters were prepared in a similar manner. We characterized the oil content and the composition of the resulting biodiesels using GC FID and HPLC. Progress of transesterication reactions and purity of esters was monitored using an Agilent 1200 series HPLC with Evaporative Light Scattering Detector (ELSD) and a Restek Allure silica column (150 9 3.2 mm i.d., 5 lm) with guard column (12.5 9 4.6 mm i.d., 5 lm) as the stationary phase. The mobile phase consisted of hexanes (phase A) and acetone (phase B). Methyl esters were analyzed using isocratic conditions of 82% hexanes and 18% acetone. Alkyl esters other than FAME were analyzed using a gradient of 96%A:4%B held for 1.5 min, then to 90%A:10%B in 0.1 min, held at 90%A:10%B for 1.9 min, then to 50%A:50%B in 1.5 min, held at 50%A:50%B for 3.5 min, and nally returned to 96%A:4%B in 1 min and held at this last composition for an additional 1 min. Retention times were veried against authentic sample of fatty acid methyl and isopropyl esters as well as oleic acid, triolein, diolein, and monoolein. The fatty acid prole of the extracted oil was determined by gas chromatography using an Agilent 7890 GC with ame-ionization detector and an Agilent HP-Innowax column (30 m 9 250 lm 9 0.25 lm). The oven program was set at 170 C for 2 min, ramped from 170250 C at 15 C/min, and held at 250 C for 8 min. Retention times and responses were veried against authentic samples of individual fatty acid methyl esters. A Differential Scanning Calorimeter (DSC) Q-20 (TA Instruments) with an autosampler was used to determine

Experimental Procedures Plant material for this study was collected from plants growing in Douglas, Bayeld and Burnett counties in Wisconsin. Solvents and alcohol reagents were purchased from Fisher Scientic (Pittsburg, NJ). Soybean oil and sodium metal were purchased from Sigma-Aldrich (St. Louis, MO). Authentic samples of fatty acid methyl esters and isopropyl esters, triolein, diolein, and monoolein were purchased from Nu Chek Prep (Elysian, MN). Oil Extraction Prior to oil extraction, seeds were dried for 8 h at 100 C. The seeds (*20 g) were ground to ne particles (0.52 mm in size) using a coffee grinder and weighed directly into Soxhlet extraction thimbles. The oil was extracted for 1415 h with 170 mL of hexanes using soxhlet extraction. Hexanes solvent was removed and recovered by rotary evaporation until a constant weight of oil was obtained. The weight of the extracted oil was then used to calculate the oil content of the seed material. Transesterication reactions have been conducted as follows: FAME Preparation A 1.60 mL volume of 1:1 (wt/wt) methanol-diethyl ether containing 0.40 mmol sodium methoxide (0.25 M) was added to a dry round-bottom ask followed by 1.00 g of the oil. The ask was tted with a water jacketed condenser and reuxed under nitrogen with stirring for 60 min. Complete reaction of triacylglycerol (TAG) was monitored by thin layer chromatography (TLC), using 90:10:1 (volume ratio) hexane:ether:acetic acid as eluent [12]. The reaction mixture was then cooled, hexane (1 mL) was added, and the solution extracted with 4 9 2 mL distilled water followed by 2 9 1 mL saturated sodium chloride solution. The hexane-biodiesel mixture was dried with anhydrous sodium sulfate and hexanes removed under vacuum until a constant weight was obtained. The resulting FAME was analyzed for purity by HPLC to ensure that no glycerol byproducts remained.

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the onset of freezing and end of melting points for each biodiesel. The onset of crystallization in the cooling DSC curves, Tc, and of the end of melting, Tm, in the heating curves have been widely used in the analysis of low-temperature properties of biodiesel (see e.g. papers [9, 11, 14 17]). The former parameter, Tc, has demonstrated good correlation with the cloud point (CP) [1], which is the leading indicator of the low-temperature performance of biodiesel. The DSC apparatus was calibrated by heating an indium reference standard through its melting transition. A refrigerated cooling system was used for reaching subambient temperature ranges. Dry nitrogen with a ow rate of 50 ml/min was used as a purge gas. Only a very limited quantity of the material (*4 mg) was needed for each analysis. Each sample was kept isothermally for 10 min at 40 C; ramped at 5 C/min to the temperature of -70 C; kept isothermally for 30 min at -70 C; and then ramped at 5 C/min back to the temperature of 25 C. Three samples of each one of the produced biodiesels were analyzed independently and the results averaged.

Fig. 1 Sample DSC scan for methyl esters of highbush cranberry

Table 2 Results of the DSC analysis of low-temperature properties of selected biodiesels Plant Ester FAME 2-Butyl Isoamyl Highbush cranberry FAME 2-Butyl Isopropyl Balsam r FAME 2-Butyl Isoamyl Isopropyl Dotted horsemint FAME 2-Butyl Isopropyl Hazelnut FAME 2-Butyl Isopropyl Tc (C) -2.9 -13.5 -5.4 -30.6 -40.8 -37.1 -12.0 -29.9 -23.4 -18.1 -17.4 -40.1 -25.2 -19.3 -30.1 -25.5 Tm (C) 3.0 -9.7 -1.9 -24.4 -34.4 -40.3 -2.0 -27.5 -23.0 -17.1 -7.8 -35.4 -18.0 -21.9 -23.6 -18.9

Results and Discussion Results of the study of the oil content and composition of the corresponding biodiesels are presented in Table 1. For comparison, the respective data for soybean oil and soybean biodiesel (SBD) are also included. Figure 1 presents a DSC run for the methyl esters of highbush cranberry. In this Figure, the onset of crystallization Tc at -30.71 C is manifested in the cooling scan by a relatively minor (DH = 1.73 J/g) exothermal peak, while the end of melting Tm (at -24.62 C) is located at the right edge of the major endothermal dip in the heating scan. Table 2 presents the results of determination of Tc and Tm for a variety of plant species. As one can see, all of the

Soybean

Table 1 Oil content and composition (weight %) for selected feedstocks

Source Oil content 19.0 62.2 C16:0 C18:0 C18:1 C18:2 C18:3

Soybean oil* American hazelnut (C. Americana) Dotted horsemint (M. Punctata) Highbush cranberry (V. Trilobum)

11.8 3.1

4.6 1.6

21.8 80.6

53.1 14.5

8.0 0.1

27.6

3.3

1.5

4.3

19.7

69.2

23.2

1.3

0.4

54.6

42.9

0.2

* Data for the composition of typical soybean oil from [19]

presented biodiesels possess better low-temperature properties than SBD as represented by the low values of Tc and Tm. Particularly noteworthy are highbush cranberry (Tc & -31 C for its methyl esters (FAME), Tc & -41 C for its 2-butyl esters), American hazelnut (Tc & -19 C for its methyl esters, Tc & -30 C for its 2-butyl esters), and dotted horsemint (Tc & -17 C for its methyl esters, Tc & -40 C for its 2-butyl esters) . It is evident from the presented data that the values of Tc and Tm for each biodiesel correlate fairly well with the oil composition of the corresponding plant. Indeed, for example, of all the species studied, FAME of highbush cranberry (Viburnum trilobum) has exhibited the lowest

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value of Tc (-31 C). An explanation for such a remarkably good low-temperature performance likely lies in the fact that the combined content of oleic (C18:1) and linoleic (C18:2) acids constitutes approximately 98% of the overall sample mass, with only about 2% left to the saturated fatty acids (C16:0, C18:0) (Table 1), much less than the over 16% saturated fatty acid content found in soybean oil. With the values of Tc being so much lower for the unsaturated methyl esters than for the esters of the corresponding middle-length saturated esters (particularly, Tc = -40.6 C for MeC18:1 vs. Tc = 24.4 C for MeC16 and 33.7 C for MeC18 [17]), the reasons for the above correlation are apparent. Total content of the unsaturated acids in the oil of American hazelnut is similar at about 95%; however, the somewhat greater amounts of saturated acids (about 5 vs. 2% for American hazelnut) increase the value of Tc (Table 2). General composition of the oil of dotted horsemint is similar to that of American hazelnut, as it contains about 95% of unsaturated acids and 5% of saturated acids. However, the ratio of the different kinds of unsaturated fatty acids in dotted horsemint is very different. While oleic acid (C18:1) prevails in American hazelnut (80.6%), it is linolenic acid (C18:3) that prevails in the dotted horsemint at 69.2%. Even though linolenic acid has even better low-temperature characteristics than oleic acid, it is likely to worsen the oxidative stability of the corresponding biodiesel.

Fig. 2 Venn diagram for Tc and Tm of 1:1 (or 1:1:1 in the diagrams center) mixtures (by weight) of biodiesels derived from the above feedstocks

According to the results of Table 1, the unsaturated fatty acids (C18:1, C18:2, C18:3) content is much higher in all of the northern plants we analyzed than in SBD resulting in biodiesels with better low-temperature characteristics. These characteristics are similar to those found for biodiesel derived from cuphea [2], where the short-chain saturated acids (mainly C10:0 [2]) dominate, also resulting in very favorable low-temperature properties. Another feature apparent from Table 2 is the superior performance of esters of branched alcohols (in comparison with methyl esters). As indicated in the introduction, this feature can be explained by the impediment introduced by those groups to the alignment of the molecules needed for crystallization [9]. A similar, but generally not as signicant, effect can be obtained by using straight medium chain alcohols in transesterication [1]. As we mentioned in the introduction, the composition of biodiesel, including both the fatty acid content and minor constituents, varies depending upon the feedstock [10], which in turn inuences its chemical and physical properties. Figure 2 presents a Venn diagram for mixtures of equal amounts (by mass) of fatty-acid methyl esters derived from oils of seeds from highbush cranberry, dotted horsemint, and American hazelnut. As one can see, for two of these mixtures (highbush cranberry and dotted horsemint, as well as American hazelnut and dotted horsemint) the values of Tc for the corresponding binary mixture is lower than that for the constituent biodiesels individually. Since, as noted above, chemical and physical properties of biodiesel are affected by its composition (including both the fatty acid content and minor constituents) which varies depending upon the feedstock, the better performance of the above binary mixtures could be generally explained by the favorable change in the composition of the corresponding binary mixtures with respect to that of the original two biodiesels. Besides mixing biodiesels produced from different feedstocks, chemical and physical properties can also be affected by mixing biodiesels obtained from a single feedstock by transesterication with different alcohols [11]. As an example, Fig. 3 presents the Venn diagram results for mixing of equal amounts of two, three, or four different biodiesels derived from highbush cranberry. We can see that the values of Tc and Tm for the corresponding admixtures are comparable to those of individual biodiesels. These results might be of some value in evaluating the potential of using a mixture of alcohols (such as fusel oil [18]) in the synthesis of biodiesels. In conclusion, we produced biodiesels from a number of northern plants, such as highbush cranberry, dotted horsemint, and American hazelnut, that possess remarkably good low-temperature properties. Further studies are planned in order to both characterize the properties of these

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J Am Oil Chem Soc Fig. 3 Venn diagram for Tc and Tm of biodiesel mixtures prepared by mixing equal amounts (by weight) of two, three, or four biodiesels derived from highbush cranberry using different alcohols. The data for Tc and Tm of the corresponding four pure biodiesels are also displayed

biodiesels more fully and to elucidate in more detail the inuence of geographic location and soils on these properties.
Acknowledgments We are grateful to Tom Markee, Nicholas Osborn, and Divya Parapati for their help. We would also like to express our deep gratitude to the Reviewers for their remarks that were useful in improving the clarity and the overall quality of the manuscript. Funding for this project has been provided by Army Research Lab under Agreement # W911NF-08-2-0029; we are also grateful to the University of Wisconsin System ARG Program for continued funding.

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