Microchemical Journal 98 (2011) 6265

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Microchemical Journal
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m i c r o c

Simple method for the determination of Cu and Fe by electrothermal atomic absorption spectrometry in biodiesel treated with tetramethylammonium hydroxide
Mirela Ghisi a, Eduardo S. Chaves b,1, Daiane P.C. Quadros b,1, Edmar P. Marques a, Adilson J. Curtius b,1, Aldala L.B. Marques a,
a b

Departamento de Tecnologia Qumica, Universidade Federal do Maranho (UFMA), 65080-040, So Lus, MA, Brazil Departamento de Qumica, Universidade Federal de Santa Catarina (UFSC), 88040-900, Florianpolis, SC, Brazil

a r t i c l e

i n f o

a b s t r a c t
A simple analytical procedure for the determination of Cu and Fe in biodiesel samples by electrothermal atomic absorption spectrometry is proposed. An aliquot of the sample is simply mixed with tetramethylammonium hydroxide (TMAH) and heated to 90 C for 5 min. Pyrolysis and atomization temperatures were optimized through pyrolysis and atomization curves. The high pyrolysis temperature adopted, of 1000 C, certainly helped minimizing potential interferences. Even though, calibration should be carried out with aqueous standard solutions in the presence of the TMAH. The detection limits (3 s, n = 10), in the sample, were quite low 15 ng g1 and 24 ng g1 for Cu and Fe, respectively. Seven biodiesel samples, produced from different raw materials, including vegetable seed, frying oil and animal fat were analyzed. Accuracy was validated by applying the recovery test to two samples, enriching the samples with two concentration levels (recoveries from 105% to 120%). The precision, expressed by the relative standard deviation was less than 3% for Cu and less than 7% for Fe. Copper could be quantied in two and Fe in three of the seven samples. The biodiesel sample from fodder turnip was especially rich in the analytes in comparison to the other samples. 2010 Elsevier B.V. All rights reserved.

Article history: Received 30 September 2010 Received in revised form 7 November 2010 Accepted 8 November 2010 Available online 13 November 2010 Keywords: Biodiesel Copper Iron TMAH ET AAS

1. Introduction Alternative fuels to those derived from petroleum are being more intensively produced and used, as the natural reserves of oil are being exhausted or are becoming too expensive and dangerous to be explored. Biodiesel, which is renewable, non-toxic and less polluting as fuel, is a very promising alternative [1,2]. The combustion of biodiesel results in less carbon monoxide, nitrous oxide, hydrocarbons and particulate materials when compared to that of diesel from petroleum, contributing for the quality of the atmosphere [2,3]. Biodiesel is produced by the transesterication of vegetable oils or animal fat, new or after use as frying oil, with an alcohol of short chain, usually methanol or ethanol, and often employing NaOH or KOH as catalyst. This reaction produces alkyl esters from long chain fatty acid and glycerin as a sub product [36]. There are several vegetable oils produced from different plants seeds in Brazil, such as babassu, palm seed, castor bean, cotton seed, curcas bean, fodder turnip, peanut, soybean, sunower and tung [7]. However, soybean is mainly used [8]. Biodiesel can be mixed in any proportion with mineral diesel to be used as fuel [9].

Corresponding author. Tel.: + 55 98 33018683. E-mail addresses: mirelaghisi@gmail.com (M. Ghisi), aldalea.ufma@hotmail.com (A.L.B. Marques). 1 INCT de Energia e Ambiente do CNPq, www.inct.cienam.ufba.br. 0026-265X/$ see front matter 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.microc.2010.11.003

Metals can be incorporated in fuels during the production process, by contact with renement or distillation equipments, storage and transport and some metallic compounds can be added to improve characteristics of the products. However, unless as additives, the presence of metallic elements is undesirable, even in low concentrations, and their determination is important to guarantee the quality of the product and to control the pollution due to their release into the atmosphere by the fuel combustion [10,11]. In case of biodiesel, the presence of metals can also be attributed to their absorption by the plant from the soil [12]. Metals in the seeds may be extracted to the vegetable oil and to biodiesel during processing [7,13]. In Brazil, the specications for biodiesel are established by the Agncia Nacional do Petrleo, Gs Natural e Biocombustveis (ANP), through Resolution no. 7 from 2008. Contaminants from the raw material, such as Ca + Mg, P and S and from the production process, such as Na + K, have maximum concentrations established by ANP [14]. For other elements there are no concentration limits in biodiesel, however, monitoring of Cu, Fe and other metals is also important to guarantee the quality of the product, as they may catalyze oxidative and degradation reactions [15,16]. A method to check the biodiesel stability using infrared spectroscopy was recently proposed [17]. It is interesting to point out that biodiesel was found to reduce the wearing of the combustible motors, reducing the amount of metallic contaminants in the emission [9]. Some methods were already proposed for the determination of elements in biodiesel by atomic spectrometry techniques. Calcium, P,

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Mg, K and Na were determined by inductively coupled plasma mass spectrometry after sample dilution in ethanol [18]. Sodium and K were determined by ame atomic emission spectrometry (F AES) after sample microemulsion formation [19]. The determination of Na, K, Ca and Mg by F AAS in biodiesel after microemulsion formation was also proposed [2022]. More sensitive techniques such as inductively coupled plasma mass spectrometry (ICP-MS) and electrothermal atomization atomic absorption spectrometry (ET AAS) have been used. In this way, the determination of Cu, Fe, Mn, Ni and V in biodiesel and diesel samples by electrothermal vaporization ICP-MS following sample emulsion preparation was proposed [23]. In addition, the determination of Cd, Co, Cu, Mn, Ni, Pb, Ti, and Zn in biodiesel by ICP-MS after microemulsion sample preparation was also proposed [24]. Different calibrations techniques and modiers for the determination of low concentrations of Cd, Pb and Tl in biodiesel samples as microemulsion by ET AAS were investigated [25]. Using two sample preparation procedures, microemulsion formation and acid digestion in a microwave oven, Cu, Pb, Ni and Cd were determined in biodiesel by ET AAS [26]. Nickel and Cd in biodiesel samples were also measured by ET AAS using W as permanent modier after microemulsion sample preparation [27]. A more complete overview of the atomic spectrometry methods published in the literature can be found in a recent review on the elemental analysis of biodiesel and vegetable oils [28]. Viscous and complex samples, such as biodiesel samples, require conversion to a form compatible with the used instrumentation, allowing, if possible, a simple and effective calibration [23]. Acid digestion of oils requires the use of strong acids, in disagreement with the green chemistry principles and is usually time consuming, even if assisted by microwaves. In addition, it is subjected to analyte loss and/ or sample contamination [29]. Sample emulsication is an interesting alternative, but emulsion instability and calibration may be problematic [20,21,23,25,26]. Dilution with an organic solvent may be fast [18], but increases the detection limit, not allowing the determination of very low concentrations. Treatment of biodiesel samples with tetramethylammonium hydroxide (TMAH) is also an interesting alternative which has being employed with success in the solubilization of biological samples such as milk powder, bovine muscle, mussel tissue, sh muscle, human hair, human blood and nail samples prior to their analysis [3035]. Usually, the treatment is very simple and fast. The objective of this work is the development of a simple analytical method for the determination of Cu and Fe in biodiesel samples by ET AAS, after sample treatment with tetramethylammonium hydroxide. The method will be applied to biodiesel samples produced from different raw materials. Accuracy validation will be carried out by the recovery test. To the best of our knowledge, this is the rst time that treatment with TMAH is applied to biodiesel samples, prior to their analysis. 2. Experimental 2.1. Instrumentation The integrated absorbance corrected for the background were measured in an atomic absorption spectrometer AAnalyst 100 (Perkin Elmer, Norwalk, CT, EUA), equipped with a deuterium-arc lamp background corrector, a longitudinally heated electrothermal atomizer HGA 800 and an auto sampler AS-72, all from Perkin Elmer. The equipment was controlled by a microcomputer, using the AA Winlab program on Windows. Hollow cathode lamps from Perkin Elmer were operated at 15 mA with measurement at 324.8 nm, using a slit width of 0.7 nm for Cu and at 30 mA, with measurement at 248.3 nm, using a slit width of 0.2 nm for Fe. Aliquots of 20 L of the treated samples and of the standard solutions used for calibration were injected into the graphite furnace.

Argon with a purity of 99.996% (White Martins, So Paulo, SP, Brasil) was used as sheath and purge gas. Pyrolytically coated graphite tubes (Part Number B0109322) with pyrolytical graphite platform (Part Number B0109324) from Perkin Elmer were used in all measurements. The optimized temperature program for both analytes is shown in Table 1.

2.2. Materials and samples All reagents used were of analytical grade. The water used in all procedures was puried in a Milli-Q system (Millipore, Bedford, MA, USA) to a resistivity of 18 m cm. Nitric acid (Carlo Erba, Milan, Italy, No. 408015), puried by subboiling distillation in a quartz still (Krner Analysentechnik, Rosenheim, Germany). A 25% m/v of TMAH in methanol (Sigma, St. Louis, MO, USA, Part No. 016838/1 41707136) was used to treat the biodiesel samples. Stock solutions containing 1000 0.002 mg L1 of each analyte were prepared by diluting Titrisol ampoules (Part No. 9987 for Cu and 1.09972 for Fe) from Merck (Darmstadt, Germany). Dilutions were carried out with water. Seven different samples of biodiesel obtained from different sources: cotton seed, castor bean, fodder turnip, soybean, soy sludge, frying oil and animal fat, using alkaline catalysis were analyzed. The sample was prepared in 15 ml polyethylene ask, by mixing 500 mg of the biodiesel with 500 L of the 25% v/v TMAH solution. After mixing, the ask was heated to 90 C in a hot plate by 5 min. The solution was diluted to 5 mL with water. For the calibration curves, standard solutions containing 0; 5; 10; 20; 30 and 40 g L1 of each analyte and 2.5% m/v TMAH were used. Calibration curves in water without TMAH and analyte addition curves were also obtained. The analyte addition curves were obtained by adding 0; 5; 10; 20; 30 and 40 g L1 of each analyte to the sample treated with TMAH.

3. Results and discussion 3.1. Sample preparation Different volumes of the 25% m/v TMAH solution: 50, 100, 200, 300, 400 and 500 L were added to 500 mg of the sample, keeping the mixture at 90 3 C for 5 min before completing the volume with water to 5 mL. For most of the samples, the highest tested volume produced a clear and transparent solution, indicating complete solubilization of the sample. For lower volumes of the TMAH solution, the nal solution was opaque. However, even using the highest tested volume of the reagent, the solution was not clear for castor oil, soy sludge and cotton seed biodiesels, but the nal slurry was enough stable and homogeneous to allow its measurement. The heating time of 5 min was considered optimum, as shorter times produced opaque solutions and times from 10 to 20 min produced a gelatinous mixture.

Table 1 Optimized temperature program for the determination of Cu and Fe in biodiesel samples using ET AAS and treatment with TMAH. Step Dry Dry Dry Pyrolysis Cooling Atomization Cleaning Temperature (C) 90 150 300 1000 20 2300 2400 Ramp time (s) 10 8 10 10 1 0 1 Hold time (s) 10 10 15 15 3 5 3 Ar ow rate (mL min1) 250 250 250 250 250 0 250

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M. Ghisi et al. / Microchemical Journal 98 (2011) 6265 Table 2 Figures of merit for the determination of Cu and Fe in biodiesel samples. Calibration (540 gL1) Cu No TMAH 2.5% v/v TMAH Analyte addition Fe No TMAH 2.5% v/v TMAH Analyte addition
a

3.2. Temperature program The temperature program was optimized through pyrolysis and atomization curves for aqueous analyte standard solutions containing 1% v/v nitric acid and for the fodder turnip or the soy sludge oils treated with TMAH, shown in Fig. 1. Similar curves were obtained for both solutions, indicating similar behavior. For Cu, the signal is quite stable up to 1200 C, while for Fe the stability remains up to a pyrolysis temperature of 1100 C. Pyrolysis at 1000 C was selected for both analytes, as this high temperature should be enough to reduce the presence of the matrix during atomization. An atomization temperature of 2300 C was adopted. As the concentration of Fe in the fodder turnip oil was too high, the curves for Fe were obtained with the soy sludge biodiesel. 3.3. Validation studies Three calibration strategies were applied: calibration using aqueous standard solutions without and with 2.5% m/v TMAH and the analyte addition technique. As shown in Table 2, the slope of the curve obtained with standard solutions in the presence of TMAH were closer to the slopes of the analyte addition curve slopes, indicating that the adopted calibration should be carried out in the presence of TMAH, used to treat the samples. Without TMAH, the slopes of the external curves were about 15% higher than in presence of 2.5% m/v of this reagent. As also shown in the Table, all calibration curves had linear correlation coefcients (R) higher than 0.999, indicating good linearity. The limit of detection (LD) and the limit of quantication (LQ) were dened as the ratios 3 s/b and 10 s/b, respectively, where s is the standard deviation of 10 measurements of the blank and b is the slope of the calibration curve. The obtained limits for each analyte and for the three calibration strategies, shown in Table 2, are of the same order of magnitude, being lower for the calibration in the absence of TMAH. The limits for the adopted calibration, in the presence of TMAH, and for the analyte addition calibration are about the same, around 15 ng g1 for Cu and around 24 ng g1 for Fe in the biodiesel samples. Recently, the National Institute of Standards and Technology (NIST) has introduced two certied reference materials for biodiesel (NIST 2772 and NIST 2773); however, the concentrations of Fe are not certied and the informed values are below their detection limits. As

Slopea (s L g1) 0.0142 0.0006 0.0119 0.0001 0.0109 0.0004

LD (ng g1) 12 15 16

LQ (ng g1) 37 46 49

0.9997 09999 0.9991

0.0169 0.0007 0.0151 0.0007 0.0152 0.0008

0.9999 0.9993 0.9999

22 24 24

67 73 73

Slope condence interval at 95%.

these materials cannot be used for accuracy validation, the recovery test was applied for two biodiesel samples, enriching the samples with two different concentrations of Cu and Fe. The obtained results, shown in Table 3, demonstrate the accuracy of the proposed method, considering that the recoveries were in the range 105%120%. 3.4. Analytical application Seven biodiesel samples, obtained from different raw materials were analyzed, after treatment with TMAH and calibration in the presence of this reagent. The obtained concentrations results are shown in Table 4. The concentration ranges for both analytes are quite wide, going from less than 46 ng g1 to 182 ng g1 for Cu and from less than 73 ng g1 to 4940 ng g1 for Fe. Copper could be quantied only in two biodiesel samples, while Fe could be measured in three of the seven samples. The much higher concentrations of Cu and Fe in the fodder turnip can be attributed to the favorable absorption of metals from the soil by this plant. This plant is also used for soil remediation due to its absorption characteristics [36,37]. The RSD values for Cu were below 3% and for Fe were below 7%, showing adequate precision. It is interesting to compare our simple proposed procedure to that used at NIST, where the analysis was carried out by ICP OES, after a long and complex sample acid digestion, involving nitric and perchloric acids, as described in the Certicate of Analysis for NIST 2772. In addition, analyte addition calibration was used for Fe, which is also time consuming. Their detection limits, 0.2 mg kg1 for Cu and Fe, is about one order of magnitude higher than the detection limits of the proposed procedure. However, at NIST, several other trace elements were measured in the digested sample, while our work was focused on Cu and Fe. 4. Conclusions The proposed method is precise and accurate, using calibration curves obtained with aqueous standard solution containing the same amount of TMAH used for the sample treatment. As a high pyrolysis temperature was adopted, most of the matrix must be eliminated before atomization, avoiding interference, as it was demonstrated by

Table 3 Concentration of Cu and Fe (ng g1) in biodiesel samples and in biodiesel samples after spiking (115 and 230 ng g1 of the analytes). Sample Cu 115 ng g1 Fig. 1. Pyrolysis and atomization curves for Cu ( ) and for Fe ( ) in aqueous standard solution containing 0.4 pg of the analytes and for Cu () in soy sludge biodiesel and for Fe ( ) in fodder turnip biodiesel, both samples treated with 2.5% m/v TMAH. Atomization temperature for the pyrolysis curve: 2300 C and pyrolysis temperature for the atomization curves: 800 C. Soybean Frying oil Recovery (%) 128 2 128 8 111 230 ng g1 262 1 269 1 114 117 Fe 115 ng g1 130 5 121 8 105 113 230 ng g1 276 9 244 4 106 120

Without enrichments the determined analytes concentration in soybean and frying oil sample were lower than the LODs.

M. Ghisi et al. / Microchemical Journal 98 (2011) 6265 Table 4 Obtained concentrations (average condence interval at 95%, n = 3) for Cu and Fe in biodiesel samples. Sample Cu Concentration (ng g1) Cotton seed Soy sludge Castor bean Fodder turnip Frying oil Animal fat Soybean b 46 130 4 b 46 182 11 b 46 b 46 b 46 RSD (%) 2 3 Fe Concentration (ng g1) b 73 86 6 1350 160 4940 650 b 73 b 73 b 73 RSD (%) 3 6 7

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the recovery test. In addition, the treatment with TMAH avoids sample contamination and analyte loss, being simple and fast, as no digestion is required. The analytes concentrations found in the samples were especially high for the biodiesel from fodder turnip. For some samples, their concentrations were below the quantication limits. Certainly, other analytes can be determined in biodiesel using the proposed sample treatment procedure. Acknowledgements The authors are thankful to Conselho Nacional de Desenvolvimento Cientco e Tecnolgico-CNPq, Coordenao de Aperfeioamento de Pessoal de Nvel Superior-CAPES (Projeto PROCAD 211/ 2007) and Petrleo Brasileiro S/A-PETROBRAS for the nancial support. M. Ghisi, E.S. Chaves and A.J. Curtius have scholarship from CNPq. D.P.C. Quadros has scholarship from CAPES. References
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