Print version ISSN 0101-2061
Ciênc. Tecnol. Aliment. vol.31 no.1 Campinas Jan./Mar. 2011
Influência do extrato de marcela (achyrocline satureioides) na oxidação lipídica de salames
Paulo Cezar Bastianello Campagnol*; Leadir Lucy Martins Fries; Nelcindo Nascimento Terra; Bibiana Alves dos Santos; Ariane Schmidt Furtado; Edsom Roberto Lorenci Toneto; Rogério Manoel Lemes de Campos
Universidade Federal de Santa Maria - UFSM, CEP 97105-900, Santa Maria - RS, Brasil, E-mail: email@example.com
The effect of two levels (0.5 and 1%) of hydroalcoholic extract of Achyrocline satureioides on the safety (TBARS values) and quality (pH, water activity, colour, weight loss, and sensorial attributes) of salami was evaluated. The addition of Achyrocline satureioides extract decreased TBARS values significantly during the storage of salami when compared to the control, which was elaborated without Achyrocline satureioides extract. The treatment with 1% of "Marcela" extract showed larger lipid stability than that of the lot with 0.5%, However, it presented a decrease (p < 0.05) in the sensorial acceptance. The two levels of "Marcela" extract did not influence pH, water activity, colour, and weight loss significantly. This study indicates that the hydroalcoholic extract of "Marcela" was effective in decreasing the lipid oxidation and at 0.5% it did not alter the sensorial features; therefore, it may be used in salami to provide safer products for the consumers.
Keywords: salami; lipid oxidation; natural antioxidant; "Marcela".
O efeito de dois níveis (0,5% e 1%) de extrato hidroalcoólico de marcela (Achyrocline satureioides) na segurança (valores de TBARS) e qualidade (pH, atividade de água, cor, perda de peso e atributos sensoriais) de salames foi avaliado. A adição de extrato de marcela diminuiu significativamente os valores de TBARS durante o armazenamento dos salames, comparado ao controle, elaborado sem extrato de marcela. O tratamento com 1% de extrato de marcela mostrou uma maior estabilidade lipídica que o lote com 0,5%, porém apresentou uma diminuição (p < 0,05) na aceitação sensorial. Os dois níveis de extrato de marcela não influenciaram significativamente os parâmetros de pH, atividade de água, cor e perda de peso. Este estudo indicou que o extrato hidroalcoólico de marcela foi efetivo na diminuição da oxidação lipídica e que a concentração de 0,5% não alterou as características sensoriais, podendo, portanto, ser utilizada em salames para proporcionar produtos mais seguros aos consumidores.
Palavras-chave: salame; oxidação lipídica; antioxidante natural; marcela.
The shelf life of fermented meat sausages is not usually limited by microbial deterioration, but by chemical or physical alterations. Due to the high percentage of fat and low water activity, lipid oxidation is considered one of the main limiting shelf-life factors of those products.
Lipid oxidation causes modifications of organoleptic features of meat products, such as, for instance, alterations of meat colour and fat, development of unpleasant taste, and smell and decrease in the nutritional value of the product due to the decrease in the content of polyunsaturated fatty acids, whose beneficial effect on consumers' health is well documented (ALEXANDER, 1998; ROSE; CONNOLLY, 1999; BERRA; MONTORFANO; RIZZO, 2005). In addition, some intermediate and final products of oxidation reactions are potentially toxic to human health, such as the oxidation compounds of cholesterol (KUBOW, 1990; PANIANGVAIT et al., 1995) and the polymerization of triglycerides (ALEXANDER, 1978; CHANG; PETERSON; HO, 1978) besides the aldehydes with α and β instaurations including the malonaldehyde, which is known for its toxic, mutagenic and carcinogenic effects (NEWBURG; CONCON, 1980).
The antioxidants present themselves as an alternative to prevent the oxidative deterioration of foods and minimize the oxidative damages in humans. Since the application of synthetic antioxidant in food industry has been target of queries as for its innocuousness, scientific studies have been focused on the search for natural compounds which present this functional property (MELO; GUERRA, 2002).
Extensive research has been focused on discovering new sources of natural antioxidants to be used in meat and meat products (CHEAH; HASIM, 2000; MCCARTHY et al., 2001; KARPINSKA; BOROWSKI; DANOWSKA-OZIEWICZ, 2001; KANATT; CHANDER; SHARMA, 2004; KANATT et al., 2005; CAMPOS et al., 2007). Achyrocline satureioides, commonly known as "Marcela", is a medicinal plant used in Argentina, Uruguay, Paraguay, and Brazil for its antispasmodic, hepatoprotecting, and coleretic properties. The high content of polyphenolic compounds, most of them flavonoids, and different phenolic acids such as capheic, chlorogenic, and isochlorogenic (SIMÕES et al., 1988; FERRARO; NORBEDO; COUSSIO, 1981) suggests that this plant may have powerful antioxidant effects. There are some reports on the antioxidant properties of "Marcela" (GUGLIUCCI; MENINI, 2002; DESMARCHELIER; COUSSIO; CICCIA, 1998), but its application in meat products has not been studied.
This objective of this study was to evaluate the effect of "Marcela" extract (Achyrocline satureioides) on the oxidative stability and on the sensorial characteristics of fermented salami sausages during storage.
2 Material and methods
2.1 Extract preparation
The dry vegetal product (30 g of inflorescence of "Marcela") was homogenized with solvent, transferred to a beaker, and left for 1 hour at room temperature. Next, it was filtrated through filter paper (Whatman number 6). The residue was submitted to two more successive extractions with the objective of fully extract the active principle of the raw material. The 3 aliquots filtered were collected and concentrated by rotary evaporation (Rotavapor® RE 120 - Büchi, Flawil, Switzerland) until 7% of the initial volume, to obtain the crude extract which was kept under refrigeration in glass flask protected from light. In the elaboration of the extract, the liquid-solid ratio was 12:1. The solvent applied in the first extraction was a mixture of ethanol 95% with distilled water (12:1), and in the two following extractions it was ethanol 95%.
2.2 Elaboration of salami
The salami samples were elaborated according to the following formulation: pork (600 g.kg-1) and beef (300 g.kg-1), lard (100 g.kg-1), sodium chloride (25 g.kg-1), glucose (5 g.kg-1), saccharose (5 g.kg-1), commercial mixture of cure, containing nitrate and sodium nitrite (3 g.kg-1), sodium ascorbate (2.5 g.kg-1), white pepper (2 g.kg-1), garlic (3 g.kg-1), nutmeg (0.02 g.kg-1), and commercial starter culture Floracarn SPX (Chr. Hansen) (0.25 g.kg-1). Portions of pork meat and beef were ground in disks of 12 and 8 mm, respectively. Pieces of pork fat, lard, were cut in cubes of nearly 1 cm3 using a knife, After being ground, the mass of meat and the lard were added with sodium chloride and mixed in a blender for 3 minutes for the extraction of myofibrillar proteins. After adding all other ingredients the starter culture, previously diluted in chlorine-free distilled water (200 mL of water for each 100 kg of mass of meat), was finally added.
The mass of meat was divided equally into three lots, originating the following treatments: 1) control, without the addition of hydroalcoholic extract of "Marcela"; 2) addition of 0.5% of hydroalcoholic extract of "Marcela"; and 3) addition of 1% of hydroalcoholic extract of "Marcela".
The treatments were stuffed in collagen casings of 60 mm diameter and cut in slices of approximately 15 cm in length. After being stuffed, the samples were subjected to a bath in a 20% solution of potassium sorbate and ripened in a laboratory ripening cabinet (Menoncin, Erechim, Brazil) until reach water activity of 0.87.
The programming of temperature and relative humidity (Tº/R.U.%) was the following: first day, temperature 25 °C/R.U. 95%; second day, 24 °C/93%, third day, 23 °C/90%, fourth day, 22 °C/85%, fifth day, 21 °C/80%, sixth day, 20 °C/75%, and from seventh day on, 18 °C/75%. When ready, the guts were removed, and the pieces of the fermented sausages were vacuum-packed and kept at room temperature for 90 days.
2.3 Physicochemical analysis
The measurement of pH was performed by homogenizing ten grams of sample with distilled water (1:10). The pH of the homogenized sample was determined by a pH meter (Digimed electrode) (TERRA; BRUM, 1988). The pH determination was performed on days 0, 1, 2, 3, 4, 5, 6, 7, 14 and 19 of manufacturing.
Water activity determination (Aw)
The determination of the Aw was performed by using the gadget Testo 400 CE, (TESTO GMBH & CO.), and were made on days 0, 3, 7, 14, 16 and 19 of manufacturing.
The weight loss was determined by the weight difference among ten sausages per batch just after the stuffing process and after the end of sausage production (TERRA; BRUM, 1988).
The colour determination was performed with a Minolta Chroma Meter CR-300 equipment (MINOLTA). The results were expressed as L* (lightness), a* (red index), and b* (yellow index). The determinations were performed on days 0, 3, 7, 14 and 19 of manufacturing.
Evaluation of the lipidic oxidation
Thiobarbituric Acid Reactive Substances (TBARS) resulting from lipidic oxidation were determined according to Raharjo and Sofos (1993) method. The TBARS values were determined in triplicate for each sample after zero, 15, 30, 45, 60, 75, and 90 days of storage, and the results were expressed in mg of malonaldehyde for kg of sample.
2.4 Sensorial analysis
The global acceptance of the products was evaluated at zero 30, 60, and 90 days of storage with the aid of a panel constituted by 30 non-trained tasters, consumers of salami, using a structured hedonic scale of seven points varying from "I disliked it very much" (score 1) to "I liked it very much" (score 7). The samples were offered to the tasters in white plastic plates, codified with three digits, followed by a glass of water and cream crackers (MORAES, 1988).
2.5 Statistical analysis
All determinations were made in triplicate and the data were evaluated through the variance analysis (ANOVA). Means were compared by the Tukey test, considering the significance level of de 5% (p < 0.05) using the statistical pack SAS (1996), version 6.12.
3 Results and discussion
3.1 Physicochemical analysis
The changes in the pH values during the manufacturing period of the salami are presented in Figure 1. During the first seven manufacturing days, a decrease in the pH values from 5.80 to nearly 4.95 was observed, probably due to the accumulation of lactic acid formed by the lactic acid bacteria (TERRA, 1998). The decline in the pH value during the first days of fermentation is very important for the production of high quality safe salami due to the inhibition of undesirable microorganisms, conversion and stabilization of colour, and formation of desireable taste and smell compounds (LÜCKE, 1994). After the seventh day, the pH values increased up to 5.14. This may be attributed to the production of ammonia and biogenic aminates as a result of the enzymatic activity (LÜCKE, 1998; BOZKURT; ERKMEN, 2002). The "Marcela" extract did not affect significantly (p > 0.05) the pH values during the period of salami production, suggesting that its addition does not cause alterations in the fermentative process.
The changes in water activity (Aw) during the period of salami production are shown in Figure 2. Aw decreased during the production reaching the value of 0.87 in all treatments after 19 days. This reduction is due to the decrease in the pH because the water retention capacity of meat proteins decreases when the pH gets closer to its isoelectric point accelerating the dehydration and consequently reducing the Aw (CHASCO; LIZASO; BERIAIN, 1996). The statistical analysis showed that the addition of "Marcela" extract did not influence significantly (p > 0.05) the water activity during the production period.
The weight loss was 46.92, 46.78, and 45.86% in the control lot and in treatments with 0.5 and 1% of "Marcela" extract, respectively. No significant difference (p > 0.05) was observed among the treatments. Those values are very close to the range of 30 to 40%, which is considered ideal for dry fermented products (RUST, 1994).
Table 1 presents the colour parameters during the salami production. The L* values decreased in all treatments during the 19 days of production, and no significant difference was observed among the treatments. This decrease represents the formation of a dark colour due to the darkening reactions (BOZKURT; BAYRAM, 2006). Similarly, Kayaardi and Gök (2003) verified that the L* values in salami usually decrease during the maturation period.
The a* values increased during the whole production period (Table 1). During the first days of fermentation, the nitric oxide that is already present in the meat combines with myoglobin producing tnitrosomyoglobin (LÜCKE, 1994). Since this pigment has red colour, the a* values increased during the salami elaboration. The addition of "Marcela" extract reduced significantly the a* values at the beginning (0 day) and on the 3rd day of production. However, from the 7th day on, although lower in the treatments with "Marcela" extract, the a* values did not differ significantly from the control.
The b* values decreased in all treatments during the salami production, and no significant difference was observed among them (Table 1). Those results agree with the data obtained by Perez-Alvarez et al. (1999), who observed a decrease in the b* values of salami during fermentation and maturation. The authors attributed this decrease to the consumption of oxygen by microorganisms and to the consequent decrease in oxymyoglobine, which contributed to the yellow colouring
The TBARS test is the most usual method to follow the evolution of the lipidic oxidation in meat and meat products (RAHARJO; SOFOS, 1993). The changes in the TBARS values were followed during the 90 days of storage (Table 2). The lipidic oxidation was significantly affected (p < 0.05) by the addition of "Marcela" extract. At the beginning of the storage (0 day), the TBARS values of the treatments containing 0.5 and 1% of "Marcela" extract presented a decrease of 26% and 44%, respectively, in relation to the control, which presented significantly lower values than those of the control during the whole period of storage. After 45 days of storage, the TBARS number in the treatment containing 1% of "Marcela" extract was half the value found in the control. Those results suggest that the "Marcela" extract retarded the lipidic oxidation during the storage period of the salami. It was observed in this experiment, for all treatments, an increase in the TBARS values up to the 45th day of storage followed by a decrease. This decrease may be attributed to the reactions of malonaldeyde with proteins during the storage period (SAMMET et al., 2006). Nassu et al. (2003) in a study evaluating the effect of different levels of rosemary (Rosmarinus officinalis) on the oxidative stability of fermented sausage of caprine, reported TBARS values similar to those found in this study during a storage period of 90 days. Similar results have also been reported by Bozkurt (2006) in a study on the effect of different antioxidants on Turkish salami ("sucuk") during its production.
3.2 Sensorial analysis
The average scores of global acceptance as a function of storage time are presented in Figure 3. According to Labuza and Schmidl (1988), the end of the life cycle of the product occurs when there is a decrease of 1.5 points at the hedonic scale. In this experiment, this did not occur for any sample indicating that all treatments may be considered acceptable up to 90 days of storage at room temperature. No significant differences were found between the treatment containing 0.5% of "Marcela" extract and the control. However, the addition of 1% of hydroalcoholic extract of "Marcela" decreased significantly the global acceptance depreciating the sensorial quality of the product.
The hydroalcoholic extract of "Marcela" was effective in decreasing the lipidic oxidation of salami during storage. The addition of 0.5% of hydroalcoholic extract of "Marcela" did not interfere in the global acceptance of the product. Therefore, this concentration may be used in salami elaboration providing safer products for the consumers.
ALEXANDER, J. C. Biological effects due to changes in fats during heating. Journal of the American Oil Chemists' Society, v. 55, n. 10, p. 711-717, 1978. [ Links ]
ALEXANDER, J. W. Immunonutrition: The role of É-3 fatty acids. Nutrition, v. 14, n. 7-8, p. 627-633, 1998. [ Links ]
BERRA, B.; MONTORFANO, G.; RIZZO, A. M. Omega-6 e omega-3: razionale per lo studio del loro rapporto. Progress in Nutrition, v. 7, n. 1, p. 24-33, 2005. [ Links ]
BOZKURT H.; BAYRAM, M. Colour and textural attributes of sucuk during ripening. Meat Science, v. 73, n. 2, p. 344-350, 2006. [ Links ]
BOZKURT H.; ERKMEN, O. Effects of starter cultures and additives on the quality of Turkish style sausage (sucuk). Meat Science, v. 61, n. 2, p. 149-156, 2002 [ Links ]
BOZKURT, H. Utilization of natural antioxidants: Green tea extract and Thymbra spicata oil in Turkish dry-fermented sausage. Meat Science, v. 73, n. 3, p. 442-450, 2006. [ Links ]
CAMPOS, R. M. L et al. Fatty acid and volatile compounds from salami manufactured with yerba mate (Ilex paraguariensis) extract and pork back fat and meat from pigs fed on diets with partial replacement of maize with rice bran. Food Chemistry, v. 103, n. 4, p. 1159-1167, 2007. [ Links ]
CHANG, S. S.; PETERSON, R. J.; HO, C. T. Chemical reactions involved in the deep-fat frying of foods. Journal of the American Oil Chemists' Society, v. 55, n. 10, p. 718-727, 1978. [ Links ]
CHASCO, J.; LIZASO, G.; BERIAIN, M. J. Cured colour development during sausage processing. Meat Science, v. 44, n. 3, p. 203-211, 1996. [ Links ]
CHEAH, P. B.; HASIM, N. H. A. Natural antioxidant extract from galangal (Alpinia galanga) for minced beef. Journal of the Science of Food and Agriculture, v. 80, n. 10, p. 1565-1571, 2000. [ Links ]
DESMARCHELIER, C.; COUSSIO, J.; CICCIA, G. Antioxidant and free radical scavenging effects in extracts of the medicinal herb Achyrocline satureioides (Lam.) D.C. ("marcela"). Brasilian Journal of Medical and Biological Research, v. 31, n. 9, p. 1163-1170, 1998. [ Links ]
FERRARO, E. G.; NORBEDO, C.; COUSSIO, J. Polyphenols from Achyrocline satureioides. Phytochemistry, v. 20, n. 8, p. 2053-2054, 1981. [ Links ]
GUGLIUCCI, A.; MENINI, T. Three different pathways for human LDL oxidation are inhibited in vitro by water extracts of the medicinal herb Achyrocline satureioides. Life Sciences, v. 71, n. 6, p. 693-705, 2002. [ Links ]
KANATT, S. R. et al. Potato peel extract - a natural antioxidant for retarding lipid peroxidation in radiation-processed lamb meat. Journal of Agricultural and Food Chemistry, v. 53, n. 5, p. 1499-1504, 2005. [ Links ]
KANATT, S. R.; CHANDER, R.; SHARMA, A. Effect of irradiated chitosan on the rancidity of radiation-processed lamb meat. International Journal of Food Science and Technology, v. 39, n. 9, p. 997-1003, 2004. [ Links ]
KARPINSKA, M.; BOROWSKI, J.; DANOWSKA-OZIEWICZ, M. The use of natural antioxidants in ready-to-serve food. Food Chemistry, v. 72, n. 1, p. 5-9, 2001. [ Links ]
KAYAARDI, S.; GÖK, V. Effect of replacing beef fat with olive oil on quality characteristics of Turkish soudjouk (sucuk). Meat Science, v. 66, n. 1, p. 249-257, 2003. [ Links ]
KUBOW, S. Toxicity of dietary lipid peroxidation products. Trends in Food Science and Technology, v. 1, p. 67-71, 1990. [ Links ]
LABUZA, T. P.; SCHIMDL, M. K. Use of sensory data in the shelf life testing of foods: principles and graphical methods for evaluation. Cereals Foods World, v. 33, n. 2, p. 193-206, 1988. [ Links ]
LÜCKE, F. K. Fermented meat products. Food Research International, v. 27, n. 3, p. 299-307, 1994. [ Links ]
LÜCKE, F. K. Fermented sausages. In: WOOD, B. J. B (Org.). Microbiology of fermented foods. 2nd ed. London: Blackie Academy Professional, 1998. v. 2, p. 441-483. [ Links ]
McCARTHY, T. L. et al. Evaluation of the antioxidant potential of natural food/plant extracts as compared with synthetic antioxidants and vitamin E in raw and cooked pork patties. Meat Science, v. 58, n. 1, p. 45-52, 2001. [ Links ]
MELO, E. A.; GUERRA, N. B. Ação antioxidante de compostos fenólicos naturalmente presentes em alimentos. Boletim do SBCTA, v. 36, n. 1, p. 1-11, 2002. [ Links ]
MORAES, M. A. C. Métodos para avaliação sensorial dos alimentos. 6. ed. Campinas: UNICAMP, 1988. [ Links ]
NASSU, R. T. et al. Oxidative stability of fermented goat meat sausage with different levels of natural antioxidant. Meat Science, v. 63, n. 1, p. 43-49, 2003. [ Links ]
NEWBURG, D. S.; CONCON, J. M. Malonaldehyde concentrations in food are affected by cooking conditions. Journal of Food Science, v. 45, n. 6, p. 1681-1687, 1980. [ Links ]
PANIANGVAIT, P. et al. Cholesterol oxides in foods of animal origin. Journal of Food Science, v. 60, n. 6, p. 1159-1174, 1995. [ Links ]
PEREZ-ALVAREZ, J. A. et al. Physicochemical characteristics of Spanish-type dry-cured sausage. Food Research International, v. 32, n. 9, p. 599-607, 1999. [ Links ]
RAHARJO, S.; SOFOS, J. N. Methodology for measuring malonaldehyde as a product of lipid peroxidation in muscle tissues: a review. Meat Science, v. 35, n. 2, p. 145-169, 1993. [ Links ]
ROSE, D. P.; CONNOLLY, J. M. Omega 3 fatty acids as cancer chemopreventive agents. Pharmacology & Therapeutics, v. 83, n. 3, p. 217-244, 1999. [ Links ]
RUST, R. E. Productos embutidos. In: PRICE, J. F.; SCHWEIGERT, B. S. (Org.). Ciencia de la carne y de productos carnicos. 2nd ed. Zaragoza: Acríbia, 1994. p. 415-440. [ Links ]
SAMMET, K. et al. Assessment of the antioxidative potential of dietary supplementation with α-tocopherol in low-nitrite salami-type sausages. Meat Science, v. 72, n. 2, p. 270-279, 2006. [ Links ]
SAS Institute Inc. Cary, NC, 1996. [ Links ]
SIMÕES, C. M. O. et al. Pharmacological investigations on Achyrocline satureioides (Lam.) DC., compositae. Journal of ethnopharmacology, v. 22, n. 3, p. 281-293, 1988. [ Links ]
TERRA, N. N. Apontamentos de tecnologia de carnes. São Leopoldo: Unisinos, 1998. [ Links ]
TERRA, N. N.; BRUM, M. A. R. Carne e seus derivados: técnicas de controle de qualidade. São Paulo: Nobel, 1988. [ Links ]
Recebido para publicação em 7/11/2008
Aceito para publicação em 8/7/2009 (003938)
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