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Procedia Food Science 00 1 (2011) 000000 Procedia Food Science (2011) 1725 1731

11th International Congress of Engineering and Food (ICEF11)

Anthocyanin extraction from Jabuticaba (Myrciaria cauliflora) skins by different techniques: economic evaluation
Priscilla C. Veggi, Diego T. Santos, M. Angela A. Meireles*a
LASEFI/DEA/FEA (School of Food Engineering)/UNICAMP (University of Campinas), Rua Monteiro Lobato, 80; 13083-862, Campinas- SP, Brazil

Abstract Jabuticaba (Myrciaria cauliflora) is grape-like in appearance and texture, although its skin is thicker and tougher. This Brazilian fruit has a dark purple to almost black skin color due to a high content of anthocyanins that cover a white gelatinous flesh inside. Jabuticaba is well known for its antioxidant properties due to the presence of anthocyanins and other phenolic compounds. Anthocyanins pigments act as strong antioxidants and are antiinflammatory, with antimutagenic and cancer chemopreventive activities. Several technologies for the extraction of anthocyanin pigments from various plants sources have been proposed in literature. On the other hand, there is a lack of information about the operational costs of this extraction process at industrial scale. In this work different techniques were evaluated in terms of economical feasibility for extraction of anthocyanins from skins of jabuticaba. Ultrasound assisted (UAE), agitated bed (ABE), soxhlet and pressurized liquid extraction (PLE) methods were economically compared. Ethanol was used as extraction solvent for all extraction techniques. The simulation was conducted using the software SuperPro designer 6.0. According to the results, PLE resulted in higher extraction efficiency followed by UAE, soxhlet and ABE. The COM of the process was obtained in terms of global yield. PLE process was the most economically viable method to obtain extracts rich in anthocyanins.
2011 Published by Elsevier SelectionLtd. and/orSelection peer-review under responsibility of 11thunder International Congress 2011 Published byB.V. Elsevier and/or peer-review responsibility on Engineering and Food (ICEF 11) Executive Committee.

of ICEF11

Executive Committee Members

Keywords: Anthocyanin Extraction; Myrciaria cauliflora Skins; Different Techniques; Economical Evaluation

1. Introduction The desire for a healthier diet allied with the increasing concern of consumers over the use of synthetic additives in food has pushed the food industry to search for new sources of natural pigments [1]. Anthocyanins are a type of functional pigment responsible for a wide range of colors present in

* Corresponding author. Tel.: +55 19 35214033; fax: +55 19 35214027 E-mail address: meireles@fea.unicamp.br.

2211601X 2011 Published by Elsevier B.V. Selection and/or peer-review under responsibility of 11th International Congress on Engineering and Food (ICEF 11) Executive Committee. doi:10.1016/j.profoo.2011.09.254

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vegetables, flowers, fruits, and derived products. It is known that anthocyanin pigments act as strong antioxidants and are anti-inflammatory, with antimutagenic and cancer chemopreventive activities [2]. These bioactive properties have already been demonstrated in in vitro and in vivo studies [3], and an increase of publications in this field has been observed in recent years. In a recent review paper, Santos and Meireles [4] compiled the recent studies on the health-promoting properties of anthocyanins. This review demonstrated that consumption of dietary phytochemicals, of which anthocyanins represent a considerable part, may promote several health benefits: reduction in the risk of cardiovascular diseases, diabetes and cancer; a protective effect against hepatic and gastric damage and collagen degradation; an increase of cognitive performance, etc. Grape peels, grape by-products (constituted mainly by peels) and berries are well known for their antioxidant properties due to the presence of anthocyanins and other phenolic compounds. Many studies have been done to extract and evaluate these compounds on the industrial scale. In Brazil another source seems promising; jabuticaba (Myrciaria cauliflora) is grape-like in appearance and texture, although its skin is thicker and tougher. This fruit has a dark purple to almost black skin color due to a high content of anthocyanins that cover a white gelatinous flesh inside [4]. 2. Materials & Methods 2.1. Plant Material Jabuticaba fruits (Myrciaria cauliflora) harvested from a plantation in the State of So Paulo, Brazil, were acquired from a fruit and vegetable market center (CEASA-Campinas, Brazil). Immediately after acquisition, the fruits were stored in the dark in a domestic freezer (-10C) (Double Action, Metalfrio, So Paulo, Brazil) until sample preparation. Before extraction, the fruits were manually peeled. 2.2. Extraction Procedures 2.2.1. Ultrasound Assisted Extraction (UAE) Jabuticaba skins were added to 50-cm3 Erlenmeyer flasks and then mixed with different volumes of ethanol 99.5 % (Ecibra, Santo Amaro, Brazil) to give a feed to solvent ratio of 1:10. Immediately after the addition of the solvent, the flasks were sonicated in an ultrasonicator bath with a 40-kHz frequency (81 W) (model T 1440, Thornton, So Paulo, Brazil) at room temperature for 2 hours. After ultrasound extraction, the solvent was separated from the plant residue by simple filtration and evaporated using a rotary evaporator (Laborota, model 4001, Vertrieb, Germany), with vacuum control (Heidolph Instruments Gmbh, Vertrieb, Germany) and a thermostatic bath at 40C. The extracts were stored (- 10C) in the dark until analysis. 2.2.2. Agitated Bed Extraction (ABE) The extraction was carried out at 30C by placing jabuticaba skins into 125-cm3 Erlenmeyer flasks containing ethanol (99.5%, Ecibra, Santo Amaro, Brazil) using a feed to solvent ratio of 1:10. Extractions were carried out in a shaker (model MA 420, Piracicaba, Brazil) with agitation (150 rpm) for 2 hours. After extraction, the solvent was separated from the plant residue and evaporated, and the extract was stored as described before.

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2.2.3. Soxhlet Extraction Approximately 25 g of jabuticaba skins and 250 cm3 of ethanol or acidified ethanol (acidified to pH 3 with HCl) (99.5% Ecibra, Santo Amaro, Brazil) were used (feed to solvent ratio of 1:10). The extraction was done in a soxhlet apparatus for 8 hours, and after that the solvent was evaporated, and the extract was stored as described before. 2.2.4. Pressurized Liquid Extraction (PLE) In the pressurized liquid extraction the solvent was pumped by a HPLC pump (Thermoseparation Products, Model ConstaMetric 3200 P/F, Fremoni, USA) into the extraction cell placed in an electrical heating jacket at a desired temperature until the required pressure was obtained. All connections within the system were achieved with stainless steel tubes (1/16 and 1/8). Based on our previously experiments extraction pressure, temperature, static extraction time and solvent flow rate were set at 50 bar, 80 C, 9 min and 1.67 mL/min (during 12 min). After PLE, anthocyanin extracts were rapidly cooled to 5 C in ice water to prevent anthocyanin degradation. Subsequently, the extraction cell was exaustively purged with a flow rate of 0.71 kg/h of carbon dioxide 99.9% (Gama Gases Especiais Ltda., Campinas, Brazil) during 8-9 min to ensure that no residual anthocyanin extract solution would be into the extraction cell. At the end, ethanol from the extract solution was evaporated, and the extract was stored as described before. 2.3. Process Simulation SuperPro Designer 6.0 was used for process simulation. This software allows the mass and energy balance estimation for all streams of the process, estimates purchase costs, and reports stream and equipment data, as well as capital and manufacturing costs. The extraction denoted in this paper by agitated bed extraction (ABE) was developed using the software in a similar manner to the work of Takeuchi et al. (2009), as shown in Figure 1. The extraction procedure consists in placing a known mass of jabuticaba skins immersed in a known volume of solvent inside an agitated tank. The equipment consists of two extractors, extract-solution tank, pump, evaporator, condenser, and recycled solvent tank. The presence of the second extraction vessel among the equipment permits the simulation of a continuous process: while one of the vessels is under operation, the other one goes through the cleaning and recharging processes. For simulating UAE extraction, in accordance with a possible setup of an ultrasound extraction reactor described by Vinatoru (2001), it was assumed that ultrasonic transducers are bonded to tank external walls. Then, during the extraction process, the raw material stays immersed in the solvent inside an agitated tank receiving the ultrasound treatment. Moreover, the excessive energy dissipation in the form of heat may lead to the degradation of the substrate and hence requires cooling; for this reason, the final temperature in the cooling operation step was 30C. To simulate soxhlet extraction, some modifications were made to the process, as shown in Figure 2. The process also consisted of placing a known mass of jabuticaba skins immersed in a known volume of solvent inside an agitated tank, and the solution is heated to 78C. The presence of a condenser connected to the extractor simulates the condensation step of solvent extraction and its reflux to the extractor. The reflux operation is exhaustedly repeated.

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Priscilla al. / Procedia Food Science 1 (2011) 1725 000000 1731 P. C. Veggi, D. T. Santos, C. M.Veggi A. A. et Meireles/ Procedia Food Science 00 (2011)

Fig. 1. Flowchart of Agitated Bed Extraction (ABE) developed in SuperPro Designer

All authors must sign the Transfer of Copyright agreement before the article can be published. This transfer agreement enables Elsevier to protect the copyrighted material for the authors, but does not relinquish the authors' proprietary rights. The copyright transfer covers the exclusive rights to reproduce and distribute the article, including reprints, photographic reproductions, microfilm or any other reproductions of similar nature and translations. Authors are responsible for obtaining from the copyright holder permission to reproduce any figures for which copyright exists. The extraction denoted in this paper by pressurized liquid extraction (PLE) was developed using the software as shown in Figure 3. The PLE process developed is formed by a solvent storage tank (ethanol), pump, two extractors (while one of the vessels is under operation, the other one goes through the cleaning and recharging processes) that operate semi-continuous, the flow controller and a distiller. 2.4. Economical Evaluation The estimation of the cost of manufacturing (COM) was done for the crude extract obtained by UAE, ABE, soxhlet and PLE. The main costs that compose the COM are similar to the ones described by Turton et al. [5], which are given by total capital investment cost and operating cost. The total capital investment cost represents the fixed capital investment (FCI), working capital and start-up cost. The first one involves expenses with equipment, installation, territorial taxes, engineering, etc., while the second one represents operating liquidity available to a business, and finally, the start-up cost is associated with the beginning of operation and the validation of the process. The operating cost represents direct costs that are directly dependent on the production rate; it is composed of the cost of raw materials (CRM), the cost of the lost solvent during the process, utilities cost (CUT), which represents the demand for steam and cooling water required for the evaporator and condenser, electricity, and operational labor cost (COL). 2.5. Scale-up The scale-up procedure assumed that the industrial scale unit has the same performance as the laboratorial scale unit when the solvent to feed ratio between the mass of solid and solvent are kept constant (S/F), as well the true density of substrate and operational conditions. The process was designed to run 7920 h per year, which corresponds to 330 days per year with continuous 24-h per day shifts. This study considered an industrial setup with extractors of 0.05, 0.1 and 0.3 m3. The amount of jabuticaba skins used per industrial batch immersed in ethanol was determined for each capacity. The solvent loss was considered to be 10% of the total ethanol involved in the process. The true density of the

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vegetable material was 1450.5 kg/m3. The amount of ultrasonic transducers needed to each capacity in the UAE process and, consequently, the price of them was estimated using information provided by Unique Group (Indaiatuba, Brazil).

Fig. 2. Flowchart of soxhlet extraction developed in SuperPro Designer

Fig. 3. Flowchart of pressurized liquid extraction (PLE) developed in SuperPro Designer

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3. Results & Discussion The COM obtained for the extract from jabuticaba skins extraction in UAE, ABE, soxhlet and PLE processes for extractor capacities of 0.05, 0.1 and 0.3 m3 can be observed in Table 1.
Table 1. COM for extracts global yield estimated for UAE, ABE, soxhlet and PLE Extraction Technique UAE Extractor Capacity (m3) 0.05 0.10 0.30 ABE 0.05 0.10 0.30 soxhlet 0.05 0.10 0.30 PLE 0.05 0.10 0.30 13.01 9.92 9.01 11.93 Global yield (%) COM for Crude Extract (US$/kg) 794.46 530.22 401.21 1016.88 666.50 422.18 3020.00 1800.00 778.42 19.26 17.24 15.53

In general, it can be observed that the COM is inversely proportional to the extractor capacity. When the extractor capacity is raised, the COM diminishes, representing an advantage for the investment on a large industrial scale. For all extraction processes, the COM decreased between 67% and 50% when the extractor capacity was raised from 0.05 to 0.1 m3 and between 50% and 25% when it was raised from 0.05 to 0.3 m3. According to the results, PLE resulted in higher extraction efficiency followed by UAE, soxhlet and ABE. Table 1 also shows that the soxhlet extraction presented higher COM of all extraction methods used (US$ 778.50/kg for 0.3m3), probably due to its extraction time (8 hrs). The fact that the soxhlet processes is time-consuming leads to a high expense of energy and making the process economically expensive. On the other hand PLE process presented the lowest COM (15.53 US$/kg for 0.3m3). The use of Pressurized Liquid Extraction (PLE) technique is well known to be an attractive alternative, since it allows fast extraction and small solvent consumption [6]. Then the use of less solvent and time makes this process economically attractive for anthocyanin extraction from jabuticaba skins. The major advantage of PLE over conventional solvent extraction methods conducted at atmospheric pressure is that pressurized solvents remain in a liquid state well above their boiling points, allowing for high-temperature extraction. These conditions improve analyte solubility and the desorption kinetics from the matrices [7]. Hence, extraction solvents including ethanol that are inefficient in extracting phytochemicals at low temperatures may be much more efficient at elevated PLE temperatures [6]. Specifically, anthocyanin extract from jabuticaba is not available in the market. On the other hand, one can find glycolic extract of jabuticaba, which costs US$ 9.90/kg (Sarfarm, So Paulo, Brazil). A better

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comparison would require information about if there are anthocyanins in glycolic extract, an information that is not available. 4. Conclusion According to the results, PLE resulted in higher extraction efficiency followed by UAE, soxhlet and ABE. PLE process also was the most economically viable method to obtain extracts rich in anthocyanins due to the use of less solvent and time. Acknowledgements P. C. Veggi thanks FAPESP for the PhD (08/10986-2) assistantship; D. T. Santos thanks FAPESP for the post-doctoral fellowship (2010/16485-5). The authors acknowledge the financial support from FAPESP. References
[1] Montes C., Vicrio I. M., Raymundo M., Fett R., Heredia F. J. Application of tristimulus colorimetry to optimize the extraction of anthocyanins from Jaboticaba (Myricia jaboticaba Berg.). Food Res. Int. 2005;38(8-9):983-988. [2] Kong J., Chia L., Goh N., Chia T., Brouillard R. Analysis and biological activities of anthocyanins. Phytochemistry. 2003;64(5):923-933. [3] Galvano F., La Fauci L., Lazzarino G., Fogliano V., Ritieni A., Ciappellano S., Battistini N.C., Tavazzi B., Galvano G. Cyanidins: metabolism and biological properties. J. Nutr. Biochem. 2004;15(1):2-11. [4] Santos D. T., Meireles M. A. A. Jabuticaba as a Source of Functional Pigments. Phcog. Rev. 2009;3(5):127-132. [5] Turton R., Bailie R. C., Whiting W. B., Shaeiwitz J. A. Analysis, Synthesis and Design of Chemical Process, Prentice HallPTR, New Jersey. 2003. [6] Ju Z. Y., Howard L. R. Effects of Solvent and Temperature on the Extraction of Colorant from Onion (Allium cepa) Skin using Pressurized Liquid Extraction. J. Agric. Food Chem. 2003;51:5207-5213. [7] Richter B. E., Jones B. A., Ezzell J. L., Porter N. L. Accelerated solvent extraction: a new technique for sample preparation. Anal. Chem. 1997;68:1033-1039.

Presented at ICEF11 (May 22-26, 2011 Athens, Greece) as paper FPE524.