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Evaluation of the Concentration of Nonessential and Essential Elements in Chicken, Pork, and Beef Samples Produced in Brazil a
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Bruno Lemos Batista , Denise Grotto , Maria Fernanda Hornos Carneiro & Fernando Barbosa Jr.
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Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil
To cite this article: Bruno Lemos Batista , Denise Grotto , Maria Fernanda Hornos Carneiro & Fernando Barbosa Jr. (2012) Evaluation of the Concentration of Nonessential and Essential Elements in Chicken, Pork, and Beef Samples Produced in Brazil, Journal of Toxicology and Environmental Health, Part A: Current Issues, 75:21, 1269-1279, DOI: 10.1080/15287394.2012.709439 To link to this article: http://dx.doi.org/10.1080/15287394.2012.709439
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Journal of Toxicology and Environmental Health, Part A, 75:1269–1279, 2012 Copyright © Taylor & Francis Group, LLC ISSN: 1528-7394 print / 1087-2620 online DOI: 10.1080/15287394.2012.709439
EVALUATION OF THE CONCENTRATION OF NONESSENTIAL AND ESSENTIAL ELEMENTS IN CHICKEN, PORK, AND BEEF SAMPLES PRODUCED IN BRAZIL Bruno Lemos Batista, Denise Grotto, Maria Fernanda Hornos Carneiro, Fernando Barbosa, Jr.
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Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil Food safety is a global concern. Meat represents the most important protein source for humans. Thus, contamination of meat products by nonessential elements is a ready source of human exposure. In addition, knowledge of the concentration of essential elements is also relevant with respect to human nutrition. The aim of the present study was to determine the concentration of 17 elements in pork, beef, and chicken produced in Brazil. Meat samples were analyzed by inductively coupled plasma mass spectrometry. The estimated daily intake for nonessential elements including arsenic (As), cadmium (Cd), lead (Pb), mercury (Hg), and antimony (Sb) through meat consumption is below the toxicological reference values. However, high levels were detected for the nonessential element cesium (Cs), mainly in beef samples, an observation that deserves future studies to identify the source of contamination and potential adverse consequences.
components of bone (e.g., Ca, P, Mg), second messengers (e.g., Ca), acid–base balance regulators (e.g., Na, K), and participants in redox reactions (e.g., Mn, Fe, Cu, Se), and and in maintenance of cellular pH and electrical gradients (e.g., Na, K) (Boreiko 2010; Boyes, 2010; Deveau 2010; Fleet et al. 2011; Stern 2010). Livestock production is one of the main activities of global agribusiness. According to the U.S. Department of Agriculture (USDA, 1999), world beef consumption increased approximately by 175% from the early 1960s to 2010. In 2006, about 60 million tonnes (carcass weight equivalent, CWE) of beef was consumed globally; of these, 6.9 million tonnes CWE was consumed in Brazil. Brazil has the second largest cattle herd in the world with approximately 200 million head (MAPA, 2011). Brazil is also responsible for 15.5% of world
Proteins are extremely important for body growth and development. Construction of muscle, skin, hair, and organs and production of hormones, red blood cells, and enzymes are among the several essential functions of proteins. Beef, fish, pork, and chicken, as well as milk and eggs, are the main sources of proteins for humans. However, these foods may contain harmful compounds, which, after being ingested, may reach the bloodstream and adversely affect human health. In order to ensure adequate intake of essential elements and prevent exposure to harmful elements following meat consumption, it is important to frequently control food quality. Critically important to life, the essential elements act as enzyme cofactors (e.g., Zn, Cu, Fe), organic molecule stabilizers (e.g., Zn, Cu for proteins; Mg for DNA; Co in vitamin B-12), structural
Received 8 March 2012; accepted 13 June 2012. The authors acknowledge the financial support of the São Paulo State Foundation for Scientific Research (FAPESP, Brazil) and thank the Brazilian National Council for Scientific and Technological Development (CNPq) and the Foundation for the Coordination of Improvement of Higher Education Personnel (CAPES) for fellowships. Address correspondence to Fernando Barbosa, Jr., Laboratório de Toxicologia e Essencialidade de Metais, Depto. de Análises Clínicas, Toxicológicas e Bromatológicas Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Avenida do Café s/n, Monte Alegre, 14040-903 Ribeirão Preto, SP, Brazil. E-mail:
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broiler production, of which 40.7% is exported. Furthermore, Brazil accounts for 3% of world pork production, and is the fourth largest exporter, meeting 15% of international demand (Sterman-Ferraz, & de Felício 2010). Moreover, Brazilian cattle and other livestock are raised mainly free-range. Despite these large production and consumption numbers, a systematic control of the meat quality related to contamination by xenobiotics remains a concern. Further, some investigators demonstrated contamination by nonessential elements in commercial mineral salts used for animal feeds (Marçal et al. 2002; 2003; Turra et al. 2010). Nonessential elements, which are widely distributed in the environment, exert a wide variety of adverse effects. Mercury (Hg) may occur naturally in soil, and phosphate derived from rocks may also contain nonessential elements, namely, Hg and arsenic (As) (Craw 2005; Walkuska et al. 2010). Roxarsone is a well-known feed additive that contains As. Some countries such as Austria, France, and the Netherlands do not permit As residues in chickens, and the Czech Republic prohibits concentrations higher than 0.1 µg/g (Angkanaporn et al. 2008). Despite this prohibition of As additives in animal feed (MAPA 2011), in Brazil the maximum limit for As in foods is 1 µg/g. However, studies evaluating mineral formulations in animal feed demonstrated metal contamination. Marçal et al. (2002) found cadmium (Cd) concentrations in Brazilian cattle feed mineral mixtures that were above the threshold proposed by the National Research Council (NRC 1996) (0.5 µg/g). Marçal et al. (2003) also found lead (Pb) concentrations above 30 µg/g, which is the maximum allowable concentration in cattle feed mineral mixtures in Brazil (NRC 1996). Thus, meat quality evaluation related to the presence of nonessential and essential elements is of concern for food safety and nutritional reasons. Thus, the aims of this study were to (1) determine the levels of essential and nonessential elements in confined chicken, free-range chicken, pork, and beef samples from Brazil and (2) estimate adult daily intake
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through meat consumption of each chemical element.
MATERIAL AND METHODS Chemicals and Instruments All reagents used were of analytical reagent purity. Nitric acid (HNO3 ) was distilled in subboiling stills (Kürner Analysentechnik) before use. The water (resistivity 18.2 M-cm) used in all experiments was deionized using a Milli-Q high-purity water system (Millipore RiOs-DITM, Bedford, MA). All standard calibration solutions were purchased from PerkinElmer (Shelton, CT). Triton X-100 and tetramethyl ammonium hydroxide (TMAH) 25% (w/v) in water was purchased from Sigma-Aldrich (St. Louis, MO). All the materials used to prepare solutions (bottles and Falcon tubes) were cleaned in an acid bath (10% v/v) HNO3 for 24 h. After that, they were rinsed six times with Milli-Q water and dried in a laminar flow hood. All experiments were carried out in a clean room (class 1000). Elements were determined using an inductively coupled plasma mass spectrometer (ICPMS) equipped with a reaction cell (DRC-ICPMS ELAN DRCII, PerkinElmer, SCIEX, Norwalk, CT) operating with high-purity argon (99.999%, Praxaair, Brazil). Sample introduction involved a quartz cyclonic spray chamber and a Meinhard nebulizer connected by Tygon tubes to the peristaltic pump of the ICP-MS. The instrumental settings and other operating conditions for analysis were according to Batista et al. (2009). Sample Collection and Treatment Samples were collected in 2011 in markets of different Brazilian regions (south: Porto Alegre and Frederico Westphalen cities; southeast: Belo Horizonte and Ribeirão Preto cities; and center-west: Goiânia and Cuiabá cities). The parts of the animals were selected according to consumption among the Brazilian population (IBGE, 2003). Pork cuts loin (n = 10), spare ribs (n = 10), and ham (n = 12) were selected, while beef cuts crest/chuck (n = 16),
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ELEMENTS IN BRAZILIAN FOOD PRODUCTS
rib eye steak (n = 8), rump (n = 16), and round (n = 20), and leg (n = 16) and breast (n = 20) of chicken were used. After collection, the edible parts of the samples were separated out. Tissues such as fats, skins, tendons, and bones were removed from the meat samples. Then approximately 10 g of each sample was weighed, frozen to –80◦ C, and freeze-dried (Liobrás L101, Brazil). Subsequently, samples were weighed again and percent lost in freeze-drying was calculated. The samples were then milled and sieved (406 µm pore size). The analyses were carried out according to the method developed by Batista et al. (2009), which focused on total amount of chemical elements. Prior to analysis, samples (75–100 mg) were weighed in triplicate, and 1 ml tetramethylammonium hydroxide (TMAH) 50% v/v was added. The samples were left at room temperature for 12 h for solubilization. Then the volume was made up to 10 ml with a diluent containing 0.5% v/v HNO3 and 0.01% p/v Triton X-100. In all experiments 10 µg/L of the internal standard rhodium (Rh) was used. The detection limits (3 × SD blank solution/slope) for As, Ba, Cd, Pb, Cs, Hg, Sb, Co, Mn, Se, Fe, Mg, Zn, Cu, Mo, Sr, and V were 0.0023, 0.0060, 0.0021, 0.0076, 0.003, 0.103, 0.0013, 0.0026, 0.0039, 0.024, 0.571, 0.147, 0.092, 0.019, 0.0051, 0.0042, and 0.181 µg/g, respectively. Quality Control Assessment In order to check the accuracy of the analysis, certified reference materials (RM) SRM 1577 (bovine liver), SRM 8415 (whole egg powder), and SRM 8414 (bovine muscle) from the National Institute of Standards and Technology were analyzed in each batch of ordinary sample analysis. Estimation of Daily Intake The estimated daily intake was calculated on the basis of a survey of household meat mass consumed in Brazil (IBGE, 2003) and on the concentration of elements in raw meat. The
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average percent of meat mass lost in freezedrying the meat was calculated at 72.3, 71.34, and 71.8% for pork, beef, and chicken, respectively. Thus, the estimated daily intake was calculated using the following formula: EDI = Ec × M in which EDI is the estimated daily intake of an element (mg/day/person or µg/day/person); Ec is the element concentration in raw meat; and M is the mass of raw meat consumed daily in Brazil. The estimated daily intakes for each meat type (pork, beef, or confined chicken) were calculated separately and then added together. Risk characterization for nonessential elements intake was performed based on toxicological reference values provided by the Joint Expert Committee on Food Additives (JECFA) or European Food Safety Authority (EFSA). For essential elements the results were compared to the dietary reference intakes (DRI) from the Food and Nutrition Board of the Institute of Medicine, 1997–2001 (IOM, 2002), since Brazil does not have its own dietary reference intake. Statistical Analysis Essential and nonessential element levels were reported as mean ± standard deviation (SD) and found to be nonparametric. Comparisons between the concentrations of the elements in different cuts of chicken, pork and beef were made using Mann–Whitney or Kruskal–Wallis tests, followed by Duncan’s post hoc test; p values
