IBP1942_12 LABORATORY EVALUATION FROM AFRICA AND LATIN AMERICA BY INTERLABORATORY COMPARISON OF BIOETHANOL QUALITY PARAMETERS 1 Paulo Paschoal

Borges , Janana Marques Rodrigues2, Fabiano Barbieri Gonzaga3, Mary Ane Gonalves4, Isabel Cristina Serta Fraga5, Eliane Cristina Pires do Rego6, Rodrigo Cassiano de Sena7, Lindomar Augusto dos Reis8, Renata Souza e Silva9, Dalni Mata do Esprito Santo Silva10
Copyright 2012, Brazilian Petroleum, Gas and Biofuels Institute - IBP This Technical Paper was prepared for presentation at the Rio Oil & Gas Expo and Conference 2012, held between September, 1720, 2012, in Rio de Janeiro. This Technical Paper was selected for presentation by the Technical Committee of the event according to the information contained in the final paper submitted by the author(s). The organizers are not supposed to translate or correct the submitted papers. The material as it is presented, does not necessarily represent Brazilian Petroleum, Gas and Biofuels Institute opinion, or that of its Members or Representatives. Authors consent to the publication of this Technical Paper in the Rio Oil & Gas Expo and Conference 2012 Proceedings.

Abstract
Ethanol from sugar cane is entirely produced in Brazil and it can be used as biofuel both in internal market and in diverse countries. Nowadays the control of the noxious gas from fuels is needed in order to protect our planet from greenhouse effect. Due to the huge advantages of the bioethanol fuel in comparison to the fossil fuels, specific parameters for bioethanol must be measured with metrological traceability and reliability to be consumed internationally. Because of that, the quality of the biofuel can be assured, improving the quality of the product, as well as making it acceptable by the international trade. Since 2005, Inmetro, the Brazilian Institute of Metrology has been studying the different parameters established in the specifications for bioethanol by the National Agency of Petroleum, Natural Gas and Biofuels (ANP). In order to assist the National Metrology Institutes (NMIs) and independent quality laboratories from Africa and Latin America with international acceptance and technical qualification in measurement of some quality parameters from bioethanol, Inmetro organized and coordinated an interlaboratory comparison (IC) inside the PEAAL Project named Standards for the Ethanol of Africa and Latin America. The IC has aimed to evaluate seven parameters in anhydrous ethanol that are important for the quality of this kind of fuel and are specified by each country for the trade. The z-score results obtained from this exercise for the seven parameters such as acid number, copper content, electrolytic conductivity, density, sulphate, water and ethanol contents will be shown in this paper.

1 2

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Ph.D., Metallurgical Engineer Chemical Metrology Division/Electrochemical Laboratory/Inmetro. Ph.D., Chemistry Chemical Metrology Division/Inorganic Analysis Laboratory/Inmetro. 3 Ph.D., Chemistry Chemical Metrology Division/Electrochemical Laboratory/Inmetro. 4 Master, Chemistry Chemical Metrology Division/Electrochemical Laboratory/Inmetro. 5 Ph.D., Chemistry Chemical Metrology Division/Electrochemical Laboratory/Inmetro. 6 Master, Chemistry Chemical Metrology Division/Organic Analysis Laboratory/Inmetro. 7 Ph.D., Chemistry Chemical Metrology Division/Inorganic Analysis Laboratory/Inmetro. 8 Ph.D., Chemistry Chemical Metrology Division/Inorganic Analysis Laboratory/Inmetro. 9 Master, Chemistry Chemical Metrology Division/Inorganic Analysis Laboratory/Inmetro. 10 Ph.D., Mechanical Engineer Mechanical Metrology Division/Mechanical Division/Inmetro.

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1. Introduction
Ethanol can be produced from renewable sources, such as sugarcane and corn. The products from the ethanols combustion are lower pollutants; therefore the quality of the Brazilian bioethanol due to its origin from renewable sugarcane becomes even more important. The participation of biofuel in the energetic matrix depends on a complex array of factors involving many production aspects of this energy source such as raw material and technology and also the sustainability (social, economical and environmental). For guaranteeing the quality of the ethanol so that it can be commercialized as fuel, several analytical parameters and their limit values have been established and controlled by the ANP, a Brazilian regulatory agency for oil, natural gas and biofuels (Resoluo ANP n 7, de 9.2.2011). The methods of analysis used for quality evaluation of the ethanol as fuel can be classical such as gravimetric and volumetric analysis, and instrumental. From the instrumental methods of ethanol analysis, there is predominance of three classes: chromatographic, spectroscopic and electrochemical (Stradiotto, et al. 2010). Inmetro, the Brazilian Metrology Institute, exerts a key role in the research of biofuels in general (Borges, et al., 2010; Gonalves, et al., 2011; Gonzaga, et al., 2012). Thus, Inmetro aims to contribute to fundamental research, development and production of certified reference materials (CRMs) in accordance with international procedures to provide metrological traceability for the measurements carried out by industrial laboratories. In order to assist the National Metrology Institutes (NMIs) and independent quality laboratories from Africa and Latin America with technical qualification in measurement of some parameters that define the quality of ethanol used as a fuel with international acceptance, Inmetro organized and coordinated an IC inside the PEAAL Project named Standards for the Ethanol of Africa and Latin America. This exercise was performed to compare the measurement results of the laboratories from Africa and Latin America with the reference values assigned by Inmetro for providing traceable results needed to develop reference methods and suitable reference materials in this important biofuel matrix. Thus, the IC was carried out with bioethanol samples with certified values, allowing a more confident evaluation of the performance of the laboratories regarding the determination of quality parameters for the bioethanol. This paper aims to present the results of the IC for seven quality parameters measured in anhydrous bioethanol obtained by the laboratories from Africa and Latin America: acid number, copper content, electrolytic conductivity, density, sulphate, water and ethanol contents.

2. Experimental
2.1. Sample and Characterization The comparison item was made up from samples of anhydrous bioethanol. The samples of bioethanol from sugar cane (approximately 99.6 % ethanol content) were provided by a Brazilian producer. The bioethanol was bottled in glass ampoules of 20 mL. Inmetro was responsible for the preparation, bottling, characterization, homogeneity and stability tests of the comparison items based on ISO GUIDE 35:2006. The uncertainty budget for the parameters was estimated according to GUM and Eurachem/CITAC. Seven quality parameters for the bioethanol were analysed by Inmetro to this IC: acid number, copper content, electrolytic conductivity, density, sulphate, water and ethanol contents. For acid number characterization, the measurement was carried out in triplicate by automated volumetric titration with potentiometric end-point detection. The standardization of NaOH solution was carried out by using CRM of HCl 0.01 mol kg1 from Inmetro. Copper content was determined using an ICP OES coupled to ultrasonic nebulizer and membrane desolvator. Calibration curves were prepared in anhydrous ethanol using the SRM 3114 from NIST. The performance of the methodology was evaluated by recovery test using the CRM of Cu(II) calibration solution from Inmetro. For electrolytic conductivity parameter, it was determined in duplicate at 25 C by means of a conductivity meter coupled with a cell constant of K=0.090 cm1 that was previously checked with CRM of 5 S cm1 from Inmetro. The determination of the density was carried out at 20.0 C in two replicates by the use of a digital density meter. Sulphate content was determined using a bidimensional ion-exchange chromatography system with preconcentration. All solutions were diluted gravimetrically using high purity water (tipe I) and anhydrous ethanol (9:1 w/w) at 20 C. In order to establish the traceability to the International System of Units (SI) SRM 3181 from NIST was used. Water content was determined in triplicate by coulometric Karl Fischer titration with Hydranal coulomat AG. The determination of ethanol content was performed in duplicate by using gas chromatography with flame ionization detector (GC-FID) by the on-column injection technique and propanol was used as internal standard. 2

Rio Oil & Gas Expo and Conference 2012 2.2. Homogeneity and stability studies After bottling, the homogeneity study was performed for all parameters under repeatability conditions. The homogeneity study of the batch of samples was carried out to determine the within and between-bottle variation. The data evaluation of the between-bottle homogeneity study is based on the approach described in ISO Guide 35:2006. The homogeneity was assessed using a one-way analysis of variance (Van der Veen et al., 2001). All parameters of interest were assessed for their stability. Stabilities studies are conducted (Lisinger at al., 2001) to establish both dispatch conditions (short-term stability) as well as storage conditions (long-term stability). Short-term stability studies were performed during four weeks. Two ampoules were kept at 50 C for 1, 2, 3 and 4 weeks. Another two samples were placed at the reference temperature of 4 C. At the end, the samples were analyzed under repeatability conditions. From the results of this study, Inmetro ensures that the samples are stable for all measured parameters for 4 weeks at50 C and 4 C. A long-term stability study aims at determining the behavior of the material under storage conditions. All parameters of interest were assessed for their stability. This study was carried out using the classical design and evaluated by the protocol based on ISO Guide 35:2006. In this design, the measurements were taken under within laboratory reproducibility conditions. The long-term stability study of test material was evaluated using regression analysis. The material was found to be stable under the conditions of this long-term stability study for all parameters at the reference temperature of 20 C. Short-term and long-term stability studies of test material were evaluated using regression analysis. The material was found to be homogeneous and stable for all parameters. 2.3. Interlaboratorial comparison: measurement, results and statistical evaluation The number of bioethanol samples needed for analyzing each parameter were sent to the laboratories by Inmetro, as well as some CRMs from Inmetro which should be used in specific analysis such as CRM of HCl 0.01 mol kg1 for acid number, CRM of 5 S cm1 for electrolytic conductivity and CRM of Cu(II) calibration solution for copper content. Each participant laboratory carried out the measurements by using its usual methods for analyzing the sample. The data were reported and sent to Inmetro to be analysed. The statistical analysis of the results was carried out by z-score to evaluate the performance of the participant laboratories according to Equation (1) and based on ISO/IEC 17043: 2010. The reference ( y ref ) and the variability (s) values, which is due to the uncertainty of the CRM, for each comparison item for bioethanol quality parameters were assigned by Inmetro. The value of z-score for each participant was reported and the performance of the laboratory was classified as satisfactory, unsatisfactory or questionable. The interpretation of z-scores is presented as follows: |z| 2, satisfactory result; 2 < |z| < 3, questionable result; |z| 3, unsatisfactory result.

zi =

yi yref s

(1)

Where: y ref is the reference value.

yi is the average result of specific laboratory i . s is the uncertainty of the CRM.

3. Results and Discussion

Table 1 shows each reference value with its expanded uncertainty (U) for the seven parameters measured in this IC. According to the Final Report (2011), these assigned values were determined after the studies of homogeneity, characterization and long-term stability carried out on the bioethanol samples by Inmetro. The U values for each parameter were obtained multiplying the combined standard uncertainty (uc) by the coverage factor 2. The uc was estimated from the square root of the square sum of the uncertainties from characterization (uchar), homogeneity (ubb) and long-term stability studies (ults), according to the Equation (2). Inmetro used only validated methods as well as CRMs traceable to the SI.
2 2 u = u 2 + ubb + ults char

(2) 3

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Table 1. Reference values assigned by Inmetro for the IC in anhydrous bioethanol for the PEAAL Project.

Ten laboratories participated in this IC: two laboratories from Brazil, two from Africa and six from Latin America, however not all of them measured all the parameters. As an example of the statistical analysis used to evaluate the performance of the laboratories by z-score, the measurement of acid number will be used. Complete analysis of the results to this IC can be seen at the Final Report of Interlaboratory Comparison in Anhydrous Bioethanol (2011), as mentioned on this paper in the reference section. Figure 1 shows the z-score plot for the acid number parameter obtained by the participant laboratories in this IC.
Acid Number
4,0 3,0 2,0

z-score

1,0 0,0 -1,0 -2,0 -3,0 -4,0 39 17 81 20 47 12 34 67

Laboratory

Figure 1. z-score plot for acid number parameter. From the 8 laboratories which measured the acid number, 5 laboratories presented unsatisfactory results; one laboratory presented a questionable result, and 2 laboratories presented satisfactory results. The participant laboratories presented different performances due to the use of different indicators, such as phenolphthalein (laboratories 20, 39, 47 and 81), bromothymol blue (laboratory 34), alpha-naphtolphthalein (laboratories 17 and 67) and also different standards such as ASTM D1613 (2006) or NBR 9866 (1987). The laboratory 12 used colorimetric method with phenolphthalein as indicator; however, it did not mention if the requirements of ASTM D1613 standard were followed. Moreover, the laboratory 34 did not mention which standard was followed. The performance of the results in percentage obtained by the laboratories according to the z-score for each parameter is presented in Figure 2. For the sulphate parameter, only one laboratory agreed to measure this parameter; its result was satisfactory. On the other hand, 8 laboratories measured the ethanol content parameter with all the results satisfactory.

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120%
Performance of the laboratories

Satisfactory 100% Unsatisfactory Questionable

80%

60%

40%

20%

0% Acid Number Copper Density Electrolytic Sulphate conductivity Water content Ethanol content

Figure 2. Performance of the laboratories in the IC for anhydrous bioethanol quality parameter measurements.

4. Conclusions
The IC showed that the laboratory participants from Brazil, Africa and Latin America obtained good performance in the measurement of all parameters, except for acid number parameter which more than 60% of the laboratories obtained unsatisfactory results. The participation of the laboratories in the IC is a key role for the harmonic development of CRM of bioethanol and also for the improvement of the measurement for the quality parameters to guarantee the reliability, comparability and traceability of the measurement in biofuels.

5. Acknowledgements
The authors want to thank the Institute of Metrology, Quality and Technology Inmetro for supporting this interlaboratory comparison together with the sponsor of Brazilian Agency for Cooperation-ABC from the Ministry of Foreign Affairs and also the PEP-Dimci, Program for Proficiency Testing in Scientific and Industrial Metrology from Inmetro. One of the authors (Isabel Cristina S. Fraga) wants to thank the Prometro and another author (Renata Souza e Silva) the FAPERJ for granting a research fellowships.

6. References
BORGES, P. P., I. C. S. FRAGA, MARQUES, B. S. R., DIAS, J. C., CUNHA, V. S. pH measurement in bioethanol by using different electrodes and according to international standards. Journal of ASTM International, v.7, n. 4, 2010. Available online at www.astm.org. GONALVES, M. A., GONZAGA, F. B., FRAGA, I. C. S., RIBEIRO, C. M., SOBRAL, S. P., BORGES, P. P., ROCHA, W. F. C. Evaluation study of different glass electrodes by an interlaboratory comparison for determining the pH of fuel ethanol. Sensors and Actuactors B, n. 158, p. 327-332, 2011. GONZAGA, F. B., GONALVES, M. A., SOBRAL, S. P., RIBEIRO, C. M. A new method for determining the acid number of ethanol fuel using coulometric titration. Fuel, n. 94, p. 70-74, 2012. LINSINGER, T. P. J., PAUWELS, J., VAN DER VEEN, A. M. H., SCHIMMEL, H., LAMBERTY, A. Homogeneity and Stability of Reference Materials. Accred. Qual. Assur. n. 6, p. 20-25, 2001. STRADIOTTO, N. R., ZANONI, M. V. B., FRAGA, I. C. S., BORGES, P. P., Fuel Ethanol Quality: Methods of analysis and reference materials. In: Sugarcane Bioethanol R & D for Productivity and Sustainability, L. A. B. Cortez (ed), So Paulo: Blcher, Fapesp, p. 815-827, 2010. VAN DER VEEN, A. M. H., LINSINGER. T. P. J., PAUWELS, J. Uncertainty calculations in the certification of reference materials. 2. Homogeneity study. Accred. Qual. Assur., n. 6, p. 26-30, 2001. ASTM D 1613. Standard test method for acidity in volatile solvents and chemical intermediates used in paint, varnish, lacquer, and related products, 2006. 5

Rio Oil & Gas Expo and Conference 2012 BIPM, IEC, IFCC, ISO, IUPAC, IUPAP, OIML (2008). Evaluation of measurement data Guide to the expression of uncertainty in measurement. First edition, GUM:1995 with minor corrections. Eurachem/CITAC. Quantifying uncertainty in analytical measurement, Second edition, Helsinki, 2000. Final Report of Interlaboratory Comparison in Anhydrous Bioethanol. Available at http://www.inmetro.gov.br/metcientifica/pdf/Final-Report-Anhydrous-Bioethanol-Project-PEAAL.pdf. ISO Guide 35. Reference Materials - General and Statistical Principles for Certification, 2006. Available at http://www.iso.org/iso/iso_catalogue. ISO/IEC 17043. Conformity assessment General requirement for proficiency testing. Geneva, 2010. NBR 9866. lcool Etlico Verificao da Alcalinidade e Determinao da Acidez Total. Rio de Janeiro: ABNT, 1987. Resoluo ANP N 7 (DOU 10.2.2011). Available at http://www.anp.gov.br.