IBP1943_12 THERMAL AND OXIDATIVE EVALUATION OF OIL AND METHYL AND ETHYL BIODIESEL OF MORINGA 1 Rebeca T.

Aguiar , Anallyne N. C. S. Oliveira 2, Marco A. R. Melo ³, Ielena E. M. Cavalcante 4, Clediana D. Calixto 5, Everton V. Silva 6, Antonia L. Souza 7, Raul Rosenhaim 8, Antônio G. Souza 9
Copyright 2012, Instituto Brasileiro de Petróleo, Gás e Biocombustíveis - IBP Este Trabalho Técnico foi preparado para apresentação na Rio Oil & Gas Expo and Conference 2012, realizado no período de 17 a 20 de setembro de 2012, no Rio de Janeiro. Este Trabalho Técnico foi selecionado para apresentação pelo Comitê Técnico do evento, seguindo as informações contidas no trabalho completo submetido pelo(s) autor(es). Os organizadores não irão traduzir ou corrigir os textos recebidos. O material conforme, apresentado, não necessariamente reflete as opiniões do Instituto Brasileiro de Petróleo, Gás e Biocombustíveis, Sócios e Representantes. É de conhecimento e aprovação do(s) autor(es) que este Trabalho Técnico seja publicado nos Anais da Rio Oil & Gas Expo and Conference 2012.

Abstract
It has been applied 5% of biodiesel in petroleum diesel (B5) in Brazil since 2010. So this paper aims to evaluate moringa biodiesel as an alternative feedstock in energetic matrix of Brazil. The physical, chemical, flow at low temperatures, thermal and oxidative properties of methyl (MMB) and ethyl (EMB) biodiesel from Moringa oleifera oil (MOO) was studied, once it is known that esters are prone to the auto-oxidation reaction which is facilitated by the number and the position of the fatty acid double bonds. In point of these data, MMB and EMB are in accordance with ASTM D6751 and Brazilian specification from ANP n°. 14:2012. The thermogravimetric assay shows that the mass loss begin at 219 °C, 136 °C and 115 °C to MOO, EMB and MMB due to the London interactions get weaker in this sequence. The oxidative stability by P-DSC to MOO, EMB and MMB had the oxidative induction period of 180 °C, 180 °C and 155 °C.

1. Introduction
Biodiesel consists in a mixture of long alkyl chain esters (C14-C22) synthesized through the transesterification reaction – in which vegetable oil, waste ois or animal fats reacts with short chain alcohols (methanol or ethanol). This biofuel can substitute partial or totally the petroleum diesel that contents around 75% of paraffinic structure with 10-15 carbon atoms. Biodiesel has the advantage of being “environmental friendly” due to its ester matrix do not have aromatic and sulfur volatile contents, it has low particulate materials emission and its compounds are renewable and biodegradable (Singh and Singh, 2010). In this way, Brazilian government legalized the mixture use of diesel-biodiesel in 2008 with the intention of expanding environmental politics, beginning with the B2 (2% of biodiesel and 98% of petroleum diesel) and it reached the proportion of 5% of biodiesel in diesel in 2010, whose regulation in Brazil is made by the Petroleum, Natural Gas and Biofuels National Agency (ANP, 2010, in Portuguese). This initiative evidences the numerous oleaginous that can be employed in biodiesel production, since Brazil has a vast territory with different environmental conditions that possibilities each region select the triacylglycerides that are more abundant, cheap and with the best climate adaptation. Although the ethanol biofuel has been from sugar cane and produced in large scale in Brazil, the route adopted to biodiesel synthesis is the methanolic in this country, whose alcohol is highly toxic. Thus, as well as biodiesel, Brazilian ethanol is renewable and biologically more accepted, beyond the encouraging of agricultural industry to produce both biofuels (Dantas et. al., 2007a; Bouaid et. al., 2009; Monteiro et. al., 2008). The influence of the feedstock variety and its respective esters promote alteration in physical and chemical properties of biodiesel, mainly its susceptibility to oxidation due to the unsaturation content that favors the auto-oxidation reactions whose reactive rate are 1, 41 and 98 to oleates (C18:1), linoleate (C18:2) and linolenates (C18:3), in this order.

______________________________
1 2

Industrial Chemisty – LACOM – DQ – CCEN – UFPB Chemical Engineer – CT – UFPB 3 Master, Chemistry – LACOM – DQ – CCEN – UFPB 4 Industrial Chemistry - LACOM – DQ – CCEN – UFPB 5 Master, Chemistry - LACOM – DQ – CCEN – UFPB 6 Chemistry - LACOM – DQ – CCEN – UFPB 7 Doctor, LACOM – DQ – CCEN – UFPB 8 Doctor, DEQ – CT – UFPB 9 Doctor, LACOM – DQ – CCEN – UFPB

Rio Oil & Gas Expo and Conference 2012 This behavior is justified by the number of the double bonds and its positions. Allylic positions are characteristic from oleate fatty acid and the bis-allylic ones are for the others two esters (Knothe, 2007). The auto-oxidation mechanism is reported with many forms in the literature (Dantas et. al., 2011; Santos et. al., 2011; Jin et. al., 2010). Figure 1 presents the most usual system for the auto-oxidation reaction. It begins with reactive species (I) that abstracts one hydrogen of a carbon atom that forms a free radical. This free radical in the presence of diatomic oxygen conceives a peroxide radical that removes another hydrogen atom, whose products are hydroperoxides and free radicals. The new free radical can react with the oxygen to continue the oxidative propagation or it bonds with another free radical to form stable products (Jain and Sharma, 2010; Ramalho et. al., 2011a).

Figure 1. Auto-oxidation reaction The methodologies that aim to evaluate the lipid oxidation stability are concerned in the accelerated aging that measures the temperature or time that the material remains unchanged. The thermogravimetric (TG) assays determine the mass variation of the sample while Pressurized Differential Scanning Calorimetric (P-DSC) evaluates the oxidation resistance, both techniques under a controlled program of temperature (Dantas et. al., 2007a; Freire et. al., 2009). In this context, this paper aims to evaluate the physical, chemical, cold flow, thermal and oxidative properties of the oil (MOO), methyl (MMB) and ethyl (EMB) of Moringa oleifera, whose origin is Indian and it has been easily adapted to North East of Brazil because it is resistant to drought and pests (Aguiar et. al., 2011).

2. Metodology
2.1. Extraction and Neutralization of Moringa Oil The moringa seeds were initially dried and crushed to pass through chemical extraction in Sohxlet with hexane for 6 h at 60 °C, whose mixture was evaporated. The oil yield was around 24,8% and it can be possible to extract up to 30% according to Mani et. al. (2007). Moringa oil was neutralized in accordance with the methodology proposed by Moreto and Fett (1998) to avoid the formation of soap during the transesterification reaction. 2.2. Transesterification of Moringa Oil The methanolysis and ethanolysis was carried with the molar proportion of 1:6 oil:alcohol and 1% (w/w) KOH as catalyst that was dissolved in the alcohol under stirring and subsequent addition of the oil, whose mixture was kept stirring for 1 h. After the reaction, the mixture was rested to the separation of two layers. The glycerol, alcohol in excess and catalyst were removed and the upper phase that is constituted of esters was washed in distillated water to remove the remaining alcohol and catalyst. Finally, the residual water was removed in an over for 1 h at 105 °C. 2.3. Sample Characterization The physical and chemical assays were realized in accordance to the following methodologies: acid number (ASTM D664), specific mass at 20 °C (ASTM D4052), kinematic viscosity (ASTM D445), Cold Filter Plugging Point (CFPP) (ASTM D6371), Cloud Point (CP) (ASTM D2500) and Pour Point (PP). Moringa oil composition was determined by the gas chromatography method. In this measurement, methyl esters obtained from the oil were evaluated according to Hartman and Lago [27]. This analysis was performed in a gas chromatograph coupled to a mass spectrometer GCMS-QP 2010 (Shimadzu) with Durabond DB-23 column (J & W Scientific). Helium was used as carrier gas (3 mL.min-1), operating under the following conditions: injector and detector at 250 °C and temperature programme of 170 °C for 16 min, rising to 210 °C at 2 °C.min -1 and injection sample of 1 mL with split ratio of 1:50. Fatty acid (FA) were identified by comparison with standards of the detector library and quantified by the area of each peak as a function of the total area of the peaks. TG curves were obtained from TA Instruments SDT 2960 non-isotherm mode with 10 C.min-1 heating rate, 100 -1 mL.min gas flow in synthetic air atmospher, temperature range 28-600 °C and approximately 10 mg of the sample in a platinum pan. 2

Rio Oil & Gas Expo and Conference 2012 P-DSC curves were obtained using a differential scanning calorimeter (TA Instruments DSC 2920) coupled with a pressure cell with non-isothermal mode processed in a platinum pan with almost 8.0 mg of the sample under 1400 kPa oxygen pressure with 100 mL.min-1 air flow, 10 °C.min-1 heat flow and temperature range 25-600 °C.

3. Results and Discutions
3.1. Fatty Acid Composition The fatty acid composition of moringa oil is presented in Table 1 in which is verified the majority presence of oleic acid with 64.2% and 8.5% of behenic that is the characteristic FA of moringa oil. It also has 69.4% of monounsaturated fatty acids (MUFA), 1% of polyunsaturated ones (PUFA) and 28.7% of saturated ones (SFA). The number of double bonds (NDB) of moringa oil is 71.2 which is lower than those found to jatropha, cottonseed, canola and sunflower oils that have, respectively, 115, 120, 124 and 142 (Anwar et. al., 2009). Thus, according with this composition we can suppose that the moringa oil and biodiesel may be stable under oxidation by the low presence of double bonds. Table 1. Fatty acid composition of moringa oil through gas chromatography Fatty Acid Palmitic (C16:0) Palmitoleic (C16:1) Estearic (C18:0) Oleic (C18:1) Linoleic (C18:2)  SFA ¹  PUFA ³  NU 5 % 7,0 1,7 6,9 64,2 1,0 28,7 1,0 71,2 Fatty Acid Araquidic (C20:0) Gadoleic (C20:1) Behenic (C22:0) Lignoceric (C24:0) Outros  MUFA ²  UFA 4 % 4,4 3,5 8,5 1,6 1,2 69,2 70,2

¹ Saturated Fatty Acids (C16:0 + C18:0 + C20:0 + C22:0 + C24:0), ² Monounsaturated Fatty Acids (C16:1 + C18:1 + C20:1), ³ Polyunsaturated Fatty Acids (C18:2 + C18:3), 4 Unsaturated Fatty Acids (MUFA + PUFA), 5 number of double bonds (1xMUFA + 2xC18:2 + 2xC18:3). ND: No Detection. 3.2. Physical and Chemical Parameters The physical and chemical characterizations of moringa oil and its methyl and ethyl biodiesel were described in Table 2 as well as the established limits by EN 14214 and ASTM D6751 specifications to biodiesel. Table 2. Physical and chemical properties of MOO, MMB and EMB Parameter Acid Value Kinematic Viscosity at 40 °C Specific Mass at 20 °C CFPP CP PP Unit mg KOH/g mm²/s kg/m³ °C °C °C MOO 0.29 47.7 912.1 MMB 0.37 5.2 876.0 16 19 15 EMB 0.29 6.0 867.0 16 20 13 EN 14214 0.5 (max.) 3.5-5.0 860-900 at 15 °C Country specification ASTM D6751 0.5 (max.) 1.9-6.0 Report -

The moringa oil showed low acid value after its neutralization, whose initial acidity was 4.6 mg KOH/g. This condition indicates the elimination of phospholipids, carotenes and free fatty acids that could be presented in the crude oil which would cause hydrolysis, fermentation or oxidation reactions and reducing the possibility of competing reactions such as soap formations during the biodiesel synthesis, once the transesterification method uses strong alkalis as catalyst and they can react with the free fatty acids to form soap. In relation to the flow properties, it is observed a decreasing in the kinematic viscosity of the oil to its biodiesel because the high viscosities of the oils are related with the stiffness of the glyceride molecule side of the triacylglycerides and its high molecular mass that provide greater London interactions. The same argument is applied for MMB and EMB due to ethylic biodiesel has one more carbon atom than the methylic one (Rosenhaim et. al., 2011). 3

Rio Oil & Gas Expo and Conference 2012 According to some authors (Dunn, 1999; Ramalho et. al., 2012), the best tests to describe low temperature behavior of fuel are: Cold Filter Plugging Point (CFPP), Cloud Point (CP) and Pour Point (PP). CP can be related with CFPP test because the first one measures the initial temperature of fuel solid formation and the second one, the operability limit of the fuel that is more accurate than the first because it determines the lowest temperature in which the fuel flows without restrictions and it simulates the fuel in the automotive motors. PP measures the lowest temperature which the sample flows, once its major composition has formed nuclei and paraffinic crystals (Dunn, 1999; Rashid et. al., 2008). Thus it can be observed that MMB and EMB had the same CFPP, although MMB showed approximated values for PP and CFPP. CP and PP values for MMB were next to those reported by Rashid et. al. (2008) that were 18 °C and 17 °C, respectively. Comparing MMB and EMB, CP was greater to EMB due to the increase of one carbon atom in the ester chain which facilities the cloud formation due to the higher interaction from London forces. In Europe, according to EN 14214, the maximum CFPP temperature is between -5 °C and 5 °C also in function of the season. In this condition, neither MMB nor EMB would be conform biodiesel at these countries due to the high content of SFA. However, it is in accordance with ANP Resolution n°. 14/2012 which these biofuels could be used due to this Brazilian specification requires that the maximum CFPP temperature at 19 °C. 3.3. Thermogravimetric Analysis The TG data and its curves are presented in Table 3 and Figure 2. It is observed that the thermal stability increases in the following order: MMB < EMB < MOO because the oil has greater London interactions and molecular mass, thus this explanation also is applied to compare the esters. MOO was thermal degraded in two steps: the first one is due to the volatilization/combustion of the fatty acids and the second is related to the combustion process of the polymers formed during the sample oxidation. As for the esters, whose mass loss occurred in one step, it is associated to its volatilization/combustion. Dantas et. al. (2007b) showed that ethyl corn biodiesel is more stable than methyl one, while Ramalho et. al. (2011b) had the opposite result to beef tallow biodiesel. Anwar et. al. (2009) reports the lubricant properties of some oleaginous including moringa oil which is more thermally stable than jatropha, cottonseed, canola and sunflower. It can be confirmed with jatropha oil and ethyl biodiesel results published by Freire et. al. (2009), whose temperature range to the first mass loss step was 203-396 °C and 108.9276.9 °C, respectively. The reason for this behavior could be explained by the low concentration of PUFA in moringa (1%) and 49.8% for jatropha (Anwar et. al., 2009), since the number of double bond makes esters or lipids more susceptible for the auto-oxidation reactions and it facilities its volatilization. Table 3. Thermogravimetric curves of MOO, MMB and EMB in synthetic air. Sample MOO MMB EMB Step 1st 2nd 1st 1st Temperature, °C 219.8 – 428.6 428.6 – 590.6 115 – 500 136 – 550 Peak Temperature, °C 405.5 445.8 242.7 260.0 m, % 85.5 14.5 100 100

100

MMB EMB MOO

80

Mass Variation, %

60

40

20

0 0 100 200 300 400 500 600

Temperature, °C

Figure 2. Thermogravimetric curves of MOO, MMB and EMB 3.4. Thermo-oxidative Analysis through Pressurized Differential Scanning Calorimetry (P-DSC) 4

Rio Oil & Gas Expo and Conference 2012 P-DSC curves – non isothermal mode – are illustrated in Figure 3 and it monitors the energy liberation of the oxidation reaction of moringa oil and biodiesel in function of the temperature. The oxidative induction temperature of MOO, EMB and MMB was around 180 °C,180 °C and 155 °C, respectively. The MOO value was near the obtained by Anwar e. al. (2009). According with Ramalho et. al. (2011a), comparing the accelerated oxidation techniques through Rancimat, PetroOXY and P-DSC, it was inferred that different methodologies reflect the different steps of the auto-oxidation mechanism (Figure 1), which PetroOXY and P-DSC are related to oxidative propagation step – not necessarily in the same region because the first one measures the time that 10% of the pressure drops and the second one, the oxidation induction time – and the Rancimat, the termination process due to the formation of volatile polar compounds. Therefore, P-DSC stands in relation to PetroOXY and Rancimat because it evaluate the beginning of the oxidative propagation.

350 300 250

BMM BEM ONM

Fluxo de Calor, mW

200 150 100 50 0 -50 0 50 100 150 200 250 300 350 400

Temperatura, °C

Figure 3. Oxidative curves of MOO, MMB and EMB through P-DSC.

4. Conclusions
The moringa oil, after its neutralization, provides to MMB and EMB its physical, chemical, thermal, oxidative and cold flow properties, since these parameters are directly related to fatty acid composition that has 64.2% oleic (C18:1), of which 69.4% are MUFA, 8.5% behenic (22:0) and 28.7% SFA, mainly those of long chain length such as araquidic (C20:0) and lignoceric (24:0). Thus, the physical and chemical parameters of MMB and EMB are in accordance with EN 14214 and ASTM D6751 specifications, except for CPFF that is specific of each European country, but it is conform to the Brazilian according ANP Resolution n°. 14/2012, which the maximum temperature for CPFF is 19 °C greater than those obtained to MMB and EMB that was 16 °C. The CP variation was explained by intermolecular forces that are larger to EMB than MMB, making these points higher to ethyl biodiesel. The descending thermal stability through TG was MOO > EMB > MMB, whose initial temperatures was 219.8 °C, 136 °C and 115 °C. The oxidative stabilities by P-DSC were up to 180 °C, 180 °C, 155 °C for MOO, EMB and MMB, respectively, it corresponds to propagation oxidative step. These stabilities are explained due to the low concentration of PUFA. Thus, we can infer that MMB and EMB may be alternative biodiesel in the energy matrix of Brazil.

5. Acknowledgments
The authors thank MCT/CGTS by provided support, FINEP to available sources through Covenant nº. 01.06.1021.00; Employment Project from Rede ARMAZIBIOD; CNPq and CAPES for the financial support.

8. References
AGUIAR, R. T.; SOUZA, A. G..; BOTELHO, J. R.; CARVALHO FILHO, J. R.; DANTAS, M. B. ROSENHAIM, R. Estudo físicoquímico e térmico do biodiesel etílico de Moringa oleifera LAM. In: SILVA, G. F.; BERGAMASCO, R.; MIRANDA, C. S. A.;

5

Rio Oil & Gas Expo and Conference 2012
SERAFINI, M. R. (Org.). Potencialidades da Moringa oleifera Lam: Volume I. São Cristóvão: Universidade Federal de Sergipe, 2011, p. 201-207. ISBN 978-85-7822-155-3. ANP. Agência Nacional de Petróleo, Gás Natural e Biocombustíveis. http://www.anp.gov.br/?id=470. Acessado em maio, 2012 (in Portuguese). ANWAR, Farooq; SHARMA, Brajendra K.; RASHID, Umer; ERHAN, Sevim Z. Lubricant properties of Moringa oil using thermal and tribological techniques. J Therm Anal Calorim, v. 96, p. 999–1008, 2009. BOUAID, A; MARTINEZ, M.; ARACIL, J. Production of biodiesel from bioethanol and Brassica carinata oil: Oxidation stability study. Bioresource Technology, v. 100, p. 2234-2239, 2009 DANTAS, M. B.; ALBUQUERQUE, A. R.; BARROS, A. K.; RODRIGUES FILHO, M. G.; ANTONIOSI FILHO, N. R.; SINFRÔNIO, F. S. M.; ROSENHAIM, R.; SOLEDADE, L. E. B.; SANTOS, I. M. G.; SOUZA, A. G. Evaluation of the oxidative stability of corn biodiesel. Fuel, v. 90, p. 773-778, 2011. DANTAS, M. B.; ALMEIDA, A. A. F.; CONCEIÇÃO, M. M.; FERNANDES JR, V. J.; SANTOS, I. M. G.; SILVA, F. C.; SOLEDADE, L. E. B.; SOUZA, A. G. Characterization and kinect compensation effect of corn biodiesel. Journal of Thermal Analysis Calorimetry, v. 87, p. 847-851, 2007a. DANTAS, M. B.; CONCEIÇÃO, M. M.; FERNANDES JR., V. J.; SANTOS, A. N.; ROSENHAIM, R.; MARQUES, A. L. B.; SANTOS, I. M. G.; SOUZA, A. G. Thermal and kinect study of corn biodiesel obtained by the methanol and ethanol routes. J Therm Anal Calorim, v. 87, p. 835-839, 2007b. DUNN, R. O. Thermal analysis of alternative diesel fuels from vegetable oils. JAOCS, v. 76, p. 109-115, 1999. FREIRE, L. M. S.; BICUDO, T. C.; ROSENHAIM, R.; SINFRÔNIO, F. S. M.; BOTELHO, J. R.; CARVALHO FILHO, J. R.; SANTOS, I. M. G.; FERNANDES JR, V. J.; ANTONIOSI FILHO, N. R.; SOUZA, A. G. Thermal investigation of oil and biodiesel from Jatropha curcas L. Journal of Thermal Analysis Calorimetry, v. 96, p. 1029-1033, 2009. HARTMAN, L; LAGO, R. C. Rapid preparation of fatty acid methyl ester from lipids. Lab Pract, v.2, p. 475-476, 1973. JAIN, S.; SHARMA, M. P. Stability of biodiesel and its blends: A review. Renewable and Sustainable Energy Reviews, v. 14, n. 2, p. 667-678, fev. 2011. JIN, F.; ZENG, X.; CAO, J.; KAWASAKI, K.; KISHITA, A.; TOHJI, K.; ENOMOTO, H. Partial hydrothermal oxidation of unsaturated high molecular weight carboxylic acids for enhancing the cold flow properties of biodiesel fuel. Fuel, v. 89, p. 2448-2454, 2010. KNOTHE, G. Review Some Aspects of Biodiesel oxidative stability. Fuel Processing Technology, v. 88, p. 669-677, 2007. MANI, S.; JAYA, S.; VADIVAMBAL, R. Opmization of Solvent Extraction of Moringa (Moringa oleifera) Seed Kernel Oil Using Response Surface Methodology, Food and Bioproducts Processing, v. 85,n. 4, p. 328-335, December 2007. MONTEIRO, C. R.; AMBROZIN, A. R. P.; LIÃO, L. M.; FERREIRA, A. G. Review: Critical review on analytical methods for biodiesel characterization. Talanta, v. 77, p. 593-605, 2008 MORETTO, Eliane; FETT, Roseane. Tecnologia de óleos e gorduras vegetais na indústria de alimentos, São Paulo: Varela editora e livraria LTDA, 1998. RAMALHO, E. F. S. M.; ALBUQUERQUE, A. R.; SOUZA, A. L.; BARRO, A. K.; MAIA, A. S.; SANTOS, I. M. G.; SOUZA, A. G. Use of different techniques in the evaluation of the oxidative stability of poultry fat biodiesel. Journal of Thermal Analysis Calorimetry, v. 106, n. 3, p. 787-791, 2011a. RAMALHO, E. F. S. M.; CARVALHO FILHO, J. R.; ALBUQUERQUE, A. R.; OLIVEIRA, S. F.; CAVALCANTI, E. H. S.; STRAGEVITCH, L.; SANTOS, I. M. G.; SOUZA, A. G. Low temperature behavior of poultry fat biodiesel: diesel blends. Fuel, v. 93, p. 601-605, 2012. RAMALHO, E. F. S. M.; SANTOS, I. M. G.; MAIA, A. S.; SOUZA, A. L.; SOUZA, A. G. Thermal characterization of poultry fat biodiesel. Journal of Thermal Analysis Calorimetry, v. 106, n. 3, p. 825-829, 2011b. RASHID, U.; KNOTHE, G; ANWAR, F.; MOSER, B. R. Moringa oleifera oil: A possible source of biodiesel. Bioresource Technology, v. 99, p. 8175–8179, 2008. ROSENHAIM, R.; SANTOS, N. A.; DANTAS, M. B.; BICUDO, T. C.; CAVALCANTI, E. H. S.; BARRO, A. K.; SANTOS, I. M. G.; SOUZA, A. G. Rheology and MT-DSC studies of the flow properties of ethyl and methyl babassu biodiesel and blends. J Therm Anal Calorim, v. 106, p. 501-506, 2011. SANTOS, V. M. L.; SILVA, J. A. B.; STRAGEVITCH, L.; LONGO, R. L. Thermochemistry of biodiesel oxidation reactions: A DFT study. Fuel, v. 90, p. 811-817, 2011. SINGH, S. P.; SINGH, D. Biodiesel production through the use of different sources and characterization of oils and their esters as the substitute of diesel: A review. Renewable and Sustainable Energy Reviews, v. 14, p. 200-216, 2010.

6