J Am Oil Chem Soc DOI 10.

1007/s11746-011-1951-4

ORIGINAL PAPER

Assessment of Different Measurement Methods Using 1H-NMR Data for the Analysis of the Transesterication of Vegetable Oils
D. F. Andrade J. L. Mazzei C. R. Kaiser L. A. dAvila

Received: 2 July 2011 / Revised: 22 August 2011 / Accepted: 6 October 2011 AOCS 2011

Abstract In the present work we make assessments of the correlation of equations based on 1H-NMR data for the analysis of FAME in 45 transesterication products with methanol from several vegetable oils (soybean, corn, sunower, canola, linseed, cottonseed and jatropha). The products employed in this study were of high and low yield. A comprehensive approach to the applicability, advantages and limitations of employing NMR expressions is described. A new expression to determine the degree of unsaturation of combined and free fatty acids was found to be satisfactory when compared with the classic expression. Estimates of uncertainties for classic equations, which have been used to determine yield and degree of unsaturation, were proposed in the present work. The results show that either one or both of the expressions for estimating yields could not be satisfactorily used for products from linseed oil, and also indicate the possibility that products from cottonseed oil may also be exceptions. Both the equations were found to be satisfactory for the products obtained

from the other oils (soybean, corn, sunower, canola and jatropha), considering the uncertainties proposed in the present work. Keywords Biodiesel 1H-NMR Vegetable oil methyl esters Quality control

Introduction Fatty acid alkyl esters (FAAE) are commercially produced by the alcoholysis of fats and oils, especially vegetable oils, and have been used as source materials in the manufacture of other products, particularly biodiesel. In biodiesel production, rened FAAE containing over 96.5% esters have been used directly (B100) [1, 2] or in blends with fossil diesel fuel [3]. It is necessary to control the quality of the composition of FAAE both to assure that the product is suitable for its intended use and to avoid the undesirable residues of nonconverted triacylglycerols (TAG) and their derivatives, free glycerol (G), monoacylglycerols (MAG) and diacylglycerols (DAG), which result from incomplete conversion or insufcient purication [4]. The presence of these minor components in biodiesel can lead to serious engine problems and increased hazardous emissions [4, 5]. In Europe, the quality standard for biodiesel, EN 14214, establishes maximum quantities of 0.02% G, 0.80% MAG, 0.20% DAG and TAG, and 0.25% total glycerol (free and bound) [2] in the rapeseed oil methyl ester. In the USA, the American Society for Testing and Materials (ASTM) biofuel standard, D6751, sets the upper limit of 0.02% G and 0.24% total glycerol for the soybean oil methyl ester [6]. The Brazilian National Agency of Petroleum, Natural Gas and Biofuels published specications for B100 that are in

D. F. Andrade (&) L. A. dAvila Departamento de Processos Organicos, Escola de Qumica, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, Bl. E, S/201, Ilha do Fundao, Rio de Janeiro 21949-900, Brazil e-mail: debora.franca.andrade@gmail.com J. L. Mazzei Departamento de Biofsica e Biometria, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, Blv. Vinte e Oito de Setembro 87, Vila Isabel, Rio de Janeiro 20551-030, Brazil C. R. Kaiser Departamento de Qumica Organica, Instituto de Qumica, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, Bl. A, S/605, Ilha do Fundao, Rio de Janeiro 21949-900, Brazil

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accordance with both the ASTM standards and the Brazilian Association of Technical Standards [1]. 1 H Nuclear magnetic resonance (1H-NMR) spectroscopy has been used with success in studies of fossil fuels [7] and is frequently used in biofuels for monitoring the synthesis, yield and quality of alcoholysis reactions and processes [816]. The rst reports on 1H-NMR analyses of FAAE synthesis using the transesterication of vegetable oils with alcohols focused primarily on determining the yield of progressive transesterication with methanol [8, 12] or ethanol [9, 16]. Yields have been determined using 1 H-NMR analysis by means of simple equations [8, 12, 16], building a calibration curve [9] or using an internal standard [11]. The relevant assignments of the chemical shifts in the 1H-NMR spectra of soybean and rapeseed oil and the respective fatty acid methyl esters (FAME), obtained products by reaction with methanol [8, 11, 12], as shown in Table 1, Fig. 1. Gelbard et al. [8] described the utilization of 1H-NMR for monitoring yields from the transesterication of rapeseed oil, which they estimated directly (CG) by integrating selected signals, according to Eq. 1 [8].   2 IME CG 100 1 3 ICH2 Signals from the methylene group adjacent to the carbonyl group present in all fatty ester derivatives at d 2.31 (Table 1) are one of the relevant signals chosen for integration. However, as this signal appears as a triplet, accurate measurements must be made to separate this triplet from the multiplet at d 2.03, which is related to allylic protons (Table 1). Knothe [12] monitored the progress of the transesterication of soybean oil by 1H-NMR using Eq. 2 and correlating the results with ber-optic near infrared spectroscopy.

 CK 100

5 IME 5 IME 9 IAG

 2

In both Eqs. 1 and 2, the yield is assumed to depend on the integration of the methoxyl group signal, which is typical for FAME, meaning that these equations cannot be used in the alcoholysis of oils and fats except with methanol. In addition, in order to quantify the yield from the transesterication of vegetable oils with ethanol, some works [9, 10, 16] have correlated equations based on 1 H-NMR methods with other techniques (viscosity and Fourier-transform-Raman measurements), but these equations cannot be used for transesterication with methanol. The importance of these equations in monitoring the yield of transesterication with methanol is evident from their appearance in several works [1720]. Correlations with NMR approaches have been studied in the monitoring of progressive transesterication of soybean and rapeseed oils only. Gelbard et al. [8] correlated Eq. 1 using 1H-NMR and molar weight, obtaining good agreement in the determination of the progressive transesterication of rapeseed oil. Knothe [12] compared the yields of progressive transesterication using Near-Infrared spectroscopy and the NMR approaches presented in this paper (Eqs. 1, 2). The author concluded that the approaches mutually conrmed each other based only on plotting the yield of soybean oil methyl ester against the reaction time. The minimum yield was approximately 55%. 1 H-NMR spectroscopy has also been employed to estimate some physico-chemical analysis parameters in vegetable oils and their transesterication products. The iodine value [21, 22] and average molecular mass [22, 23] can be determined in vegetable oils. Furthermore, the amounts of mono-, di- and tri-unsaturated fatty acid

Table 1 Reported assignments of chemical shifts (d) and peak area integrations in 1H-NMR spectra of soybean and rapeseed oils and their transesterication products [8, 11, 12] Protons Terminal methyl group Linolenic acid methyl group Methylenes, except the followings Methylenes b- to the carbonyl or to the olen Allylic protons Methylene group adjacent to the carbonyl group Diallyl methylenes Methoxyl groups Glyceryl methylene Olen and glycerol methine
a

Compoundsa TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME TAG, DAG, MAG, FAME FAME TAG, DAG, MAG, G TAG, DAG, MAG, FAME, G

db 0.90 (t) 0.98 (t) 1.30 (m) 1.60 (m) 2.03 (m) 2.31 (t) 2.72 (t) 3.70 (s) 4.104.40 (d) 5.30 (m)

Integrationc ITM ILAM IM IMB IMA ICH2 IDAM IME IAG ICA
c

TAG, DAG, MAG tri, di and monoacylglycerols, respectively; FAME fatty acid methyl esters, G glycerol. b s Singlet, t triplet, m multiplet. Notation adopted in the present work

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J Am Oil Chem Soc Fig. 1 Generic representation of the structures of a triacylglycerols and b the respective methyl esters derivatives (R, aliphatic chain), containing the most prominent fatty acid chains (oleic, linoleic and linolenic) found in vegetable oils and the respective 1 H-NMR assignments [8, 12]

a
4.10 - 4.40 __ H2C O

1.60 2.31 1.60 1.31

1.31 1.31 1.31 1.31 1.31 1.31 1.31 1.31

1.60 2.03 1.60 2.03 1.60 2.03

5.30 5.30 2.03 5.30 5.30 2.72 5.30 5.30 2.72

1.60 1.31 5.30 5.30

1.31 1.31

1.31

0.90 1.31 1.31

1.60 2.03 1.31

HC __ O 5.30
O

2.31 1.60

0.90 0.98 2.03

5.30 5.30 2.72

5.30 5.30

H2C

__

2.31

4.10 - 4.40

b
3.70
O

chains [14, 24] and the degree of unsaturation (DU) in these chains [11, 25] can also be estimated in FAME products that have been produced from vegetable oils. The determination of average DU has only been reported for FAME produced from soybean and olive oil [11, 18, 25, 26] from a comparison of the integration value for the methylene group adjacent to the carbonyl group (d 2.31) with that of the olen protons (adjusted for the underlying methine proton) [11]. Morgenstern et al. [11] estimated the DU (DUM) from soybean oil to be 1.52, which corresponds closely to the DU values of 1.54 and 1.49 reported in previous works. An equation representing DU according the denition given by Morgenstern et al. [11] (DUM) can be expressed as:   ICA IAG =4 DUM 3 ICH2 As can be seen, the use of 1H-NMR integration data in estimating transesterication yields and parameters associated with the quality of these products have only been evaluated for soybean, olive and rapeseed oil. However, a wide range of naturally available vegetables can be used as source materials for FAME production. For instance, there have been reports on the use of corn, sunower, linseed, cottonseed, castor, crambe, peanut, palm, olive, coconut, tung, piqui (Caryocar sp.), jatropha (Jatropha curcas), and karanj (Pungamia pinnata) oils [3, 2735]. As such, the present work establishes assessments of the correlation of equations based on 1H-NMR data for the analysis of the FAME in products from transesterication with methanol. Products from seven vegetable oils, namely soybean, corn, sunower, canola, linseed, cottonseed, and jatropha oil, were studied in view of the importance of this technique which has the potential to be applied successfully

as a tool in the validation of methods less expensive and much easier to keep in operation.

Experimental Procedures Vegetable Oils and Reagents Rened food grade soybean, corn, sunower, canola, and cottonseed oil and a commercial edible grade linseed oil were purchased from retail establishments in Rio de Janeiro state (Brazil). Jatropha oil was supplied by Greentec Laboratory (Escola de Qumica/UFRJ, Rio de Janeiro). Analytical grade anhydrous potassium carbonate, sodium chloride and sodium sulfate were purchased from Merck (Darmstadt, Germany). Analytical grade methanol and hexane (mixture of isomers) were supplied by Vetec (Rio de Janeiro, Brazil). NMR grade deuterated chloroform (CDCl3) and tetramethylsilane (TMS) were purchased from Cambridge Isotope Laboratories (Andover, USA). The oils and reagents were employed without further purication. Yield from the Transesterication of Vegetable Oils The transesterication of each vegetable oil was accomplished by heating with anhydrous methanol under reux, using potassium carbonate as a catalyst, following the procedure described by Guarieiro [17] with modications. Fifty milliliters of vegetable oil, previously weighed, was added to a 125 mL round-bottomed ask, containing potassium carbonate (3% mol), methanol (oil:methanol molar ratio of 1:3 or 1:9), a magnetic stirrer and an allihn

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reux condenser. The average molecular mass of the vegetable oil components was estimated from the likely composition of the combined fatty acids and the equation proposed by Guarieiro [17]. The reaction medium was kept under magnetic agitation and reux for 5, 10, 15, 30, and 90 min and was then cooled to room temperature. The methanol residue was removed in a rotary evaporator using a water pump. The glycerine phase (lower) containing alcohol and excess catalyst was separated by decantation in a separation funnel. The upper layer was extracted with 100 mL hexane. The hexane phase was washed with distilled water (3 9 50 mL) to remove any residual catalyst and any other persistent impurities. The hexane extraction step is required to prevent an emulsion from forming [17]. The hexane was removed in a rotary evaporator (vacuum, water pump) yielding a clear liquid FAME product of a light yellow color. The products were kept in anhydrous sodium sulfate for 2 h to remove any residual water, ltered again and then stored in a freezer (approximately -10 C) until analysis.
1

Uncertainty Estimates of the Integration Measurements Our main goal was to establish assessments of the correlation of Eqs. 1 and 2, for which purpose an estimate of their uncertainty was needed. Assuming that both equations are dependent upon the integration of selected signals, uncertainty in these equations may be calculated as the result of the propagation of uncertainty of the experimental variables. An estimate of experimental uncertainty was determined from the following expression and through derivations. Thus, a simple, hypothetical relationship (f) can be dened as the difference between the two equations that represent DU (Eqs. 3, 4): f ICA IAG =4 IDAM IMA =2 0 5

The theoretical value of function f is zero, independent of the yield of the transesterication. Thus, the f values calculated (Eq. 6) from the 1H-NMR integration data, using Eq. 5, are related to its uncertainty (df), or the deviation from its theoretical value (zero). fcalculated 0 df 6

H-NMR Method

H-NMR spectra were obtained using a Bruker DPX-200 spectrometer, in the conditions described before [7]. Samples of the vegetable oils and their FAME products were dissolved at 12 mg/mL in CDCl3. TMS was used as an internal standard. The assignment of hydrogen chemical shifts was identied following Gelbard et al. [8] and is summarized in Fig. 1, Table 1. The integration of the NMR peaks was calculated and normalized relative to the methylene group adjacent to the carbonyl group (ICH2) once that was found in the TAG, DAG, MAG and FAME. Degree of Unsaturation of the Fatty Acid Chains The degree of unsaturation (DU) of the free and combined fatty acid chains in the vegetable oils and respective products was determined using Eq. 3. We propose another expression in order to estimate this parameter using Eq. 4, which also represents the contribution of the number of olen groups in the fatty chains to 1H-NMR integration and is based on the contribution by the protons in the vicinal groups to the olen groups. IDAM IMA =2 DU ICH2 Determination of Transesterication Yield by Gelbard and by Knothe The yield of the transesterication of vegetable oils into FAME was calculated using Eqs. 1 (CG, %) and 2 (CK, %) for the products obtained in the present work. 4

Function f is an arithmetical expression based on the independent experimental data of integrations, each of which is subject to a random uncertainty (dICA, dIAG, dIDAM and dIMA). Thus, its deviation (df) can be expressed in terms of the propagation [41] of the uncertainties from the integration values in Eq. 5, as expressed by: df 2 of =oICA 2 dICA 2 of =oIAG 2 dIAG 2 of =oIDAM 2 dIDAM 2 of =oIMA 2 dIMA 2 7 The substitution of the derivatives from Eq. 5 gives: df 2 dICA 2 1=16 dIAG 2 dIDAM 2 1=4 dIMA 2 8 Assuming a general uncertainty for the integration values (dICA & dIAG & dIDAM & dIMA) and substituting them for a generic parameter (dI), Eq. 8 can be reduced to: df 2 2 5=16 dI2 9

Thus, approximating the coefcient to 2/3 and calculating the square root, the generic relationship can be assumed by approximation for an average uncertainty: dI 2=3 jdf j 10

If we substitute df from Eq. 6 and f from Eq. 5, we get Eq. 11, which is proposed in the present work as the means for expressing the uncertainty of the integrations:

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dI 2=3 j ICA IAG =4 IDAM IMA =2 j Estimate of Yield Uncertainties

11

The Gelbard et al. [8] and Knothe [12] equations (1, 2, respectively) were employed to derive a relationship that enabled their uncertainties to be calculated: dCG (Eq. 12) and dCK (Eq. 13) respectively. For this, the general uncertainty of the integration determinations Eq. 11 was assumed. h i1=2 dCG 1= IME 2 1= ICH2 2 CG dI 12 dCK 100 5 9
2 2 IME IAG

1=2 dI 13

oil was also analyzed, and its spectrum (Fig. 3) was markedly different than the spectra of the other oils, mainly because of the composition of the combined fatty acids. The absence of a signal at d 2.72 is due to the absence of linoleic and linolenic acid, while the presence of a signal at d 3.70 is assigned to the chemical shift in the carbinol hydrogen, which is located at the b-position of the olen group [37], found in ricinoleic acid, the main constituent of castor oil (Fig. 3). However, this chemical shift is also characteristic of the methoxyl groups in methyl esters, so this nding could be seen as a limitation of the use of 1 H-NMR in the characterization of FAME products from this starting oil. In fact, our data were inconclusive for the analysis of the FAME from castor oil (data not shown).
1

5 IEM 9 IAG 2

Uncertainty of the Degree of Unsaturation In order to compare the equations relative to the degree of unsaturation, the uncertainties of Eqs. 3 and 4 were deduced in the same way and are given by dDUM and dDU, respectively. 2 1=2 1 DUM dDUM dI 14 ICH2 ! 1=2 5=4 DU 2 dI 15 dDU ICH2

H-NMR Spectra of the Transesterication Products

Results and Discussion

1

H-NMR Spectra of the Starting Oils

In order to analyze the signicant differences among the vegetable oils and their respective FAME products, 1HNMR spectroscopy was used on the different oils employed as starting materials. A good resolution was obtained for all the 1H-NMR spectra, as can been seen in Fig. 2. The spectra for soybean oil presented the characteristic signals and intensities widely illustrated in other works [10, 11, 36]. Very few differences in the spectrum proles can be observed in Fig. 2. The most important difference is the relative area of the diallyl methylene signal at d 2.72, which is increased by the linoleic and linolenic acid chains, since they contain two and four diallyl methylene protons, respectively. Low signals at d 0.98, relative to the terminal methyl groups in the linolenic acid chains, can be observed in the rapeseed and linseed oil spectra (Fig. 2c, d, respectively). Palm oil was also analyzed by 1H-NMR but its spectrum is not discussed in this work because the oil was not properly converted (B2%) under the test conditions. Castor

In the literature, the determination of the yield of transesterication by 1H-NMR has only been reported for soybean and rapeseed oil [8, 12]. It would be expected that all vegetable oils would contain the same functional groups in the majority of the components and would hence yield the same collection of peaks in their NMR spectra. In the present work, a good resolution was obtained for all the 1HNMR spectra of the transesterication products, and the prole spectra of the products from soybean and rapeseed oils were clearly similar to the spectra from the other oils under study, excepting palm and castor oils, as described above. The major differences observed among the 1HNMR spectra of the resulting products and the respective starting vegetable oils were the expected increase and decrease in the signal intensities of methoxyl (d 3.70) and glyceryl methylene (d 4.104.40) protons, respectively, as seen in Fig. 4, which shows the spectra of the products from corn oil. Apart from castor oil, the oils studied in the present work did not present a signal at d 3.70, which contributed towards the determination of their yield, since this signal is used in the two NMR equations (Eqs. 1, 2). The glyceridic protons in the structures of MAG and DAG exhibited 1HNMR signals in the same region as the glyceryl methylene protons of the TAG (at d 4.104.40) in the feedstock. Correlation Between the Absolute Values Calculated Using the Two 1H-NMR Equations By using the Gelbard and Knothe equations, the yield of the transesterication was calculated for the 45 products, which were derived from seven different vegetable oils, under oil:methanol molar ratios of 1:3 and 1:9 and for 5, 10, 15, 30, and 90 min reaction times (Table 2). It is well known that the molar ratio of methanol to vegetable oil is one of the parameters that most affects the yield of FAME

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Fig. 2 1H-NMR spectra (200 MHz, CDCl3) of the vegetable oils used as starting materials in the present work: a corn, b sunower, c rapeseed, d linseed, e cottonseed, and f jatropha

[38]. This assumption is reasonable because a large excess of alcohol is what drives the equilibrium reaction of the transesterication to yield the FAME [38]. Thus, an oil:methanol molar ratio that is higher than the stoichiometry 1:3 ratio is recommended for a good yield. A molar ratio of 1:6 has been widely used in industrial processes, giving ester yields of over 98% w/w. In this work, oil:methanol molar ratios of 1:3 and 1:9 were employed with the aim of obtaining low- and high-yield products, respectively. Several sources were used to verify whether the two equations could be used for products obtained from vegetable oils other than those previously reported (soybean and rapeseed oil). In fact, the results presented in Table 2 show that yields were higher for an oil:methanol molar ratio of 1:9 than for 1:3. Reaction times were set between 5 and 90 min because this is another important variable that affects ester yield. In our data the yield

increased with the reaction time, which tallies with the literature [39, 40]. The one exception we cannot explain was the yield of the products obtained from soybean oil at an oil:methanol molar ratio of 1:3 at 15 min. In order to identify whether there was any direct relationship between the values obtained from the two yield equations, CG/CK was introduced in Table 2. It can be seen that this ratio was closer to one, the expected value, primarily when the yield values were closer to 100%. Considering that uncertainty in yield determinations is independent of the respective value, it would be expected that the highest percentage differences between CG and CK would be obtained at very low yield values, as can be seen for the six products obtained at yields lower than 20% with 1:3 oil:methanol molar ratio (Table 2). The fact that CG \ CK was constantly identied, suggests that NMR signals of triacylglycerols could reduce the accuracy of the

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J Am Oil Chem Soc Fig. 3 1H-NMR spectrum (200 MHz, CDCl3) for castor oil. The chemical shifts of ricinoleic acid and the chain characteristic of the castor oil composition are indicated. The proposed chemical shift at d 2.21 is based on the assignment for the contribution of two functional groups attached to the methylene group [37] and others are typical of fatty acid chains

Fig. 4 1H-NMR spectra (200 MHz, CDCl3) of products from corn oil at different yields: a 38%, b 46%, c 91%, and d 94%. Spectrum for corn oil is shown in Fig. 2a

results from one or both equations. When the yield was between 20 and 55%, lower differences between CG and CK were observed and the ratio was close to one (\10% difference), but an increased relationship between CG/CK could also be observed (n = 11), which might suggest that signals from the other derivative residues from the transesterication could have interfered in the yield estimate when Eqs. 1 and 2 were used. Exceptions of this kind

were not observed by Knothe [12], who compared the yields of progressive transesterication using Eqs. 1 and 2, because the minimum yield studied was approximately 55%. For the highest yield, 1:9 oil:methanol molar ratio, most of the products showed a 5% difference between CG and CK, or in other words, a CG/CK ratio of between 0.95 and 1.05. One important exception was the products from

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Table 2 Yield (%) calculated by Gelbard (CG) and Knothe (CK) equations and degree of unsaturation as dened by Morgenstern (DUM) and as dened in the present work (DU), with the respective uncertainties proposed herein, of the products obtained under different reaction times and oil/methanol molar ratios from different vegetable oils (n = 45) Molar ratio 1:9 CG/CK 0.91 0.24 0.87 0.26 1.00 0.18 0.96 0.17 1.02 0.21 1.04 0.18 0.81 0.30 0.98 0.30 1.03 0.24 0.91 0.25 1.04 0.20 1.10 0.20 0.74 0.45 0.80 0.23 0.80 0.34 1.24 0.02 1.27 0.03 1.26 0.03 (1.26 0.02) (1.24 0.03) 1.23 0.04 1.22 0.02 1.24 0.03 1.15 0.14 1.12 0.13 1.25 0.15 1.28 0.15 1.11 0.13 1.24 0.14 0.09 0.09 0.09 (0.02) 0.02 0.01 0.02 (1.21 0.04) (1.25 0.04) (0.02) 1.30 0.15 1.43 0.16 0.09 1.29 0.15 1.43 0.17 0.10 89 5 91 7 103 9 93 5 74 5 90 1 94 7 97 8 78 3 86 1 81 2 79 1 89 2 79 4 1.27 0.07 1.34 0.07 0.04 92 12 (1.36 0.05) (1.41 0.06) (0.03) 75 8 73 4 94 5 88 6 97 8 88 4 75 5 93 1 87 6 95 8 80 3 90 1 88 2 92 1 95 2 94 5 94 5 1.20 0.12 1.32 0.13 0.08 91 7 88 7 1.17 0.13 1.30 0.14 0.08 85 5 87 5 85 5 87 4 1.15 0.10 1.25 0.11 0.07 81 5 80 4 1.01 0.08 0.99 0.07 0.98 0.08 1.03 0.11 1.00 0.07 1.23 0.21 1.08 0.08 0.95 0.08 1.02 0.11 1.06 0.13 1.06 0.07 0.99 0.10 0.97 0.01 1.08 0.11 1.02 0.12 0.98 0.05 0.96 0.02 0.92 0.03 0.87 0.02 0.93 0.03 (1.28 0.02) (1.30 0.02) (0.01) 92 5 91 5 1.02 0.08 1.20 0.07 1.23 0.07 1.20 0.07 1.20 0.10 1.20 0.07 1.22 0.17 0.96 0.06 1.32 0.07 1.29 0.10 1.36 0.12 1.14 0.06 1.13 0.08 1.19 0.01 1.12 0.09 1.17 0.11 1.18 0.05 1.21 0.02 1.19 0.03 1.12 0.01 1.18 0.03 1.37 0.12 1.48 0.14 0.07 86 8 83 7 1.04 0.13 1.35 0.12 1.34 0.07 1.39 0.07 1.46 0.08 0.04 81 4 83 4 81 4 84 4 1.00 0.07 0.98 0.07 1.41 0.07 1.40 0.07 1.35 0.14 1.48 0.15 0.08 81 6 82 6 0.99 0.10 1.38 0.09 (1.47 0.08) (1.54 0.09) (0.05) 1.47 0.10 1.47 0.07 1.46 0.07 1.46 0.13 1.41 0.08 1.27 0.08 1.30 0.07 1.27 0.08 1.29 0.11 1.26 0.07 1.38 0.20 1.03 0.07 1.39 0.08 1.38 0.11 1.47 0.13 1.21 0.07 1.21 0.09 1.20 0.01 1.21 0.10 1.28 0.12 1.23 0.05 1.20 0.02 1.22 0.03 1.10 0.02 1.21 0.03 DUa M DU dI CG CK CG/CK DUM DU
a a

123
CK 32 5 15 3 40 5 40 4 39 5 44 5 12 3 30 6 37 5 32 6 47 6 50 6 42 51 52 dI 0.06 0.04 0.04 0.07 0.04 0.04 0.04 0.05 0.06 0.04 0.11 0.04 0.04 0.06 0.07 0.04 0.06 0.01 0.06 0.07 0.03 0.01 0.02 0.01 0.02

Source materials

Reaction time (min)a

Molar ratio 1:3

CG

Soybean

29 6

10 15

13 3

30

40 6

90

Corn

38 5

10

15

40 6

30

46 6

90

Sunower

10 3

10

15

29 7

30

39 7

Canola

90

29 6

10

15

49 7

30

55 8

90

Linseed

31

10

15

41

30

41

J Am Oil Chem Soc

90

J Am Oil Chem Soc 0.04 0.05 0.03

89 4

95 3

Molar ratio 1:9

93 4

linseed oil at the lowest yields (until 15 min reaction time), since these products were the ones that gave the lowest ratio values (CG/CK \ 0.95). CG/CK ratio values of over 1.05 were also observed for products obtained with a 1:9 oil:methanol molar ratio, but under different conditions for different sources. The conversion results obtained by 1H-NMR, according to the expressions by Gelbard [8] and Knothe [12] (Table 2), were statistically compared using t-test: two paired samples for means. As the critical value of the onetailed something |t| was 1.68 and the calculated value |t| was 0.75, we found that the conversion values obtained for the Gelbard [8] and Knothe [12] expressions yielded no signicant differences (P [ 0.05). This is well illustrated by the diagram of CG against CK (Fig. 5), which shows the linear relationship between the CG and CK values. Importance of the Uncertainty Estimate As shown in Table 2, the absolute DUM values were very similar to the proposed DU, independent of the sources or the FAME yield, as would be expected for accurate parameters. This similarity validates the hypothetical relationship f (Eq. 5) between their expressions, which was originally used to determine the uncertainty of the integration values and consequently of the yield and degree of unsaturation. Equations 14 studied in the present work, which were used to calculate CG, CK, DUM and DU, are arithmetical expressions based on independent experimental data for integrations and also are subject to random uncertainties. Thus, the respective uncertainties dCG, dCK,
100 90

dI

(0.02) (1.21 0.03) (1.18 0.03)

1.19 0.07

1.22 0.08

0.07 1.23 0.12 1.12 0.11 0.92 0.29

1.13 0.06

1.15 0.07

1.14 0.07

1.14 0.08

CK

102 5

105 6

CG

(0.03)

0.02

dI

(1.22 0.06)

1.19 0.04

DU

(1.17 0.05)

1.15 0.04

DUa M

0.81 0.18

90 4

0.97 0.06

CG/CK

1.14 0.05

DUM

1.19 0.05

DU

Data in brackets are values for the respective vegetable oils

80 70 60

CG/CK

CK (%)

11 2

Molar ratio 1:3

22 4

50 40 30 20 10 0 0 20 40 60 80 100 120

CK

92

20 5

Reaction time (min)a

CG

Table 2 continued

10

15

30

CG (%)
Fig. 5 Diagram of CG (%) versus CK (%); values calculated from the transesterication products (n = 45) obtained under different reaction times and oil/methanol molar ratios from seven different vegetable oils, using 1:3 (lled circle) and 1:9 (lled diamond) oil:methanol molar ratio

Source materials

Cottonseed

Jatropha

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dDUM and dDU can be given in terms of the uncertainty propagation equations [41] and using the expression dI, resulting in Eqs. 1215, respectively. The uncertainties are presented in Table 2. When the values for CG/CK are correlated to the yield of each product (in relation to CG) and the uncertainties are taken into account (Fig. 6), it can be seen that only nine of all 45 products failed to represent the expected value of one, even after uncertainty was taken into account. A rigorous analysis of these exceptions showed that six of them were products of linseed oil, which particularly indicates that the values obtained from the two expressions applied to the yield are very different from each other for these products. It was also found that two of the cottonseed oil products also had CG/CK values that were far from one, even considering the uncertainty values, which indicates that their products may also be exceptions when it comes to using one or both the equations for determining their yields. In view of these results, it can be suggested that one or both of the expressions for estimating yields may not be satisfactory for the products from linseed oil and cottonseed oil. Minor compounds other than acylglycerols, in the composition of the original oils could have contaminated the products, interfering in the NMR signals. On the other hand, the products studied from soybean, corn, sunower, canola and jatropha oils (total of 34 products) were close to one when the uncertainties proposed in this work were considered.

1.40

1.20

C G /C K

1.00

0.80

0.60

0.40 0 20 40 60 80 100

CG

1.40

1.20

C G /C K

1.00

0.80

0.60

0.40 0 20 40 60 80 100 120

Conclusion It could be expected that the relative intensities of the NMR peaks would vary only with differences in the degree of unsaturation among oils/esters due to the similar composition of the main components. However, in this study we showed that expressions designed to determine the yield of transesterication products cannot be used for products obtained from vegetable oils other than the ones usually reported in the literature. The results showed that either one or both of the expressions usually employed for yield estimates could not be satisfactorily applied to products from linseed oil. It was also found that one or both of the equations for determining yields may not be suitable for products from cottonseed oil. However, both expressions were found to be suitable for the products from soybean, corn, sunower, canola and jatropha oil when the uncertainties proposed in this work were considered. The results showed that castor oil, with its hydroxyl group, present spectra markedly different than the spectra of the other oils. The presence of a signal at d 3.70 is assigned to the chemical shift in the carbinol hydrogen (located at the b-position of the olen group) found in

CG

1.40

1.20

C G /C K

1.00

0.80

0.60

0.40 0 20 40 60 80 100 120

CG
Fig. 6 Plot of CG/CK versus CG, including the uncertainties proposed in the present work, for the products from: a soybean (circles) and corn (triangles) oils; b linseed (circles), cottonseed (triangles) and jatropha (squares) oils; c sunower (circles) and canola (triangles) oils

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ricinoleic acid, the main constituent of castor oil. This chemical shift is also characteristic of the methoxyl groups in methyl esters, so this nding could be seen as a limitation of the use of 1H-NMR in the characterization of FAME products from this starting oil. Another characteristic of the spectra of castor oil is the absence of a signal at d 2.72 due to the absence of linoleic and linolenic acid. Other techniques, such as chromatography, have been used to ascertain the biodiesel yield compared with the NMR method. However, so far just one or other of the equations has been correlated, and even in these cases uncertainties were not considered. With the presented results, better approaches can be developed to assess these correlation relationships. The expression proposed in the present work to determine the degree of unsaturation of combined and free fatty acid chains was more satisfactory than the established expression, and the results were very similar, independent of the sources and the yield of the transesterication.
Acknowledgments The authors are grateful to the Brazilian National Agency of Petroleum, Natural Gas and Biofuels through its Human Resources Program (PRH-13/UFRJ, Rio de Janeiro, Brazil), the National Council for Scientic and Technological Development (CNPq/Braslia, Brazil) and the Carlos Chagas Filho Research Support Foundation of the State of Rio de Janeiro (Faperj/Rio de Janeiro, Brazil), for the nancial support and grants they provided for this work. The authors also thank Greentec Laboratory (Escola de Qumica/UFRJ, Rio de Janeiro) for supplying the jatropha oil.

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