Quantification of Polycyclic Aromatic Hydrocarbons in Commonly Consumed Salami in Brazil

Polycyclic aromatic hydrocarbons (PAHs) are organic compounds that have been found in different food categories, and meat products can have high concentrations of PAHs, since the technological processes to which they are subjected, such as smoking, can produce several compounds. Considering the risk that these compounds can bring to the population's health, it is essential to develop an accurate and reliable method to evaluate the contamination of PAHs in products of animal origin. The objective of this study was to optimize and validate a method for the quantification of 4 PAHs (benz[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene) in salami. The methodology included saponification, liquid­liquid extraction, solid-phase purification, and quantification by ultra-high-performance liquid chromatography. The effects of saponification parameters were investigated by experimental design, whereas the model obtained by regression analysis was considered satisfactory with the dissolution solvent of potassium hydroxide providing the highest global sum of areas. In validation, the parameters studied were adequate and within European and INMETRO Guidelines limits. The evaluation of 22 samples indicated that 27% were contaminated with at least one of the 4 PAHs, and benz[a]anthracene being the prevalent one with content varying between < 1.00 and 17.58 µg/kg. Two samples showed PAHs contamination above the maximum tolerable limit in the European Commission Regulation.

Avaliação da presença de hidrocarbonetos policíclicos aromáticos em chocolates: exposição ao consumo e implicações à saúde / Evaluation of the presence of polycyclic aromatic hydrocarbons in chocolates: exposure to consumption and health implications

Introdução - Os hidrocarbonetos policíclicos aromáticos (HPAs) são compostos que apresentam ação potencialmente carcinogênica e mutogênica, que geram risco à saúde humana. Esses contaminantes são formados a partir da combustão incompleta de matéria orgânica, podendo ocorrer a formação durante o processamento dos alimentos em âmbito industrial (secagem, torra, pré-cozimento) e doméstico (fritar, assar e grelhar) pelo emprego de altas temperaturas. Para a produção do chocolate e outros produtos derivados do cacau, etapas como a secagem, torra e conchagem, são processos que utilizam altas temperaturas e essenciais para a produção de chocolates. Não existem estudos avaliando a presença e a exposição dietética por HPAs em chocolates no Brasil. Objetivo - Otimizar e validar a metodologia de extração e quantificação de 4 HPAs (benzo[a]antraceno (BaA), criseno (Cri), benzo[b]fluoranteno (BbF) e benzo[a]pireno (BaP)) em chocolates, quantificar sua presença em amostras de chocolates comerciais e estimar a exposição dietética aos HPAs a partir do consumo de chocolate. Métodos - As amostras de chocolate (ao leite, branco e amargo com porcentagens de cacau de 40%, 53%, 55%, 60% e 70%) foram obtidas no comércio da cidade de São Paulo. Foi realizada a otimização e validação da metodologia pelos parâmetros de linearidade, seletividade, limite de detecção, limite de quantificação, recuperação e repetibilidade. As amostras foram submetidas a extração líquido-líquido, extração em fase sólida e quantificadas por cromatógrafo líquido de alta eficiência com detector de fluorescência. A estimativa de exposição dietética (DEs) para benzo[a]pireno foi avaliada para três categorias: 1) Sexo (feminino e masculino); 2) Grupos etários: adolescentes (14 a 18 anos), adultos (19 a 59 anos) e idosos (>= 60 anos); 3) Local de residência (urbana e rural). Resultados - O método obteve faixa de linearidade entre 0,50 e 5,00 µg.kg-1 para os quatro HPAs. Com relação à seletividade, houve interferência da matriz apenas para o BaP. O método também mostrou acurácia, com recuperações médias variando entre 95,25 a 108,27% e repetibilidade mostrando valores entre 0,14 e 7,25%. Benzo[a]pireno foi encontrado em todos os chocolates, variando entre 1,58 a 4,34 µg.kg-1 de gordura. As amostras de chocolate com 53-60% cacau e chocolate 70% cacau apresentaram as maiores taxas de contaminação para BaP; já as amostras de chocolate 40% cacau e chocolate ao leite apresentaram menor contaminação. Quanto a exposição dietética, as mulheres (por gênero) e os adolescentes (por idade) apresentaram maiores DEs para BaP pelo consumo de chocolate, principalmente para o chocolate 70% cacau. Os resultados apresentaram baixa contaminação por HPAs nos chocolates analisados, no entanto, o consumo aumentado por diferentes faixas etárias pode contribuir para a DEs ao BaP. Conclusões - A metodologia padronizada foi considerada seletiva e sensível, garantindo a eficiência do ensaio. As amostras analisadas apresentaram resultados abaixo dos limites estabelecidos pelo Regulamento da Comunidade Européia No. 835/2011 para o BaP e para a soma dos 4 HPAs. Frente aos grupos analisados (sexo, idade e local de residência), os chocolates 70% cacau contribuíram para a maior exposição dietética à BaP. A presença dos quatro HPAs prioritários nos chocolates analisados demonstrou a importância de programas de monitoramento da cadeia produtiva em produtos de cacau (massa de cacau, manteiga de cacau e cacau em pó) e ingredientes adicionados (açúcar e leite em pó).

Introduction - Polycyclic aromatic hydrocarbons (PAHs) are compounds that have a potentially carcinogenic and mutogenic action, which generate risk to human health. These contaminants are formed from the incomplete combustion of organic matter, which can occur during industrial (drying, roasting, pre-cooking) and domestic (frying, roasting and grilling) processing of food by the use of high temperatures. For the production of chocolate and other products derived from cocoa, steps such as drying, roasting and conching are processes that use high temperatures and are essential for the production of chocolates. There are no studies evaluating the presence and dietary exposure to PAHs in chocolates in Brazil. Objective - To optimize and validate the methodology for the extraction and quantification of 4 PAHs (benzo[a]anthracene (BaA), chrysene (Chr), benzo[b]fluoranthene (BbF) and benzo[a]pyrene (BaP)) in chocolates, quantify the presence in commercial chocolate samples and estimate dietary exposure to PAHs from the consumption of chocolate. Methods - Chocolate samples (milk, white and dark with cocoa percentages of 40%, 53%, 55%, 60% and 70%) were obtained from commercial stores in the city of São Paulo. The methodology was optimized and validated by the parameters of linearity, selectivity, limit of detection, limit of quantification, recovery and repeatability. The samples were subjected to liquid-liquid extraction, solid-phase extraction and quantified by a High Performance Liquid Chromatograph with fluorescence detector. The dietary exposure estimate (DEs) for benzo[a]pyrene was assessed for three categories: 1) Gender (female and male); 2) Age groups: adolescents (14 to 18 years), adults (19 to 59 years) and elderly (>= 60 years); 3) Place of residence (urban and rural). Results - The method obtained a linearity range between 0.50 and 5.00 µg.kg-1 for the four PAHs. Regarding selectivity, there was interference from the matrix only for BaP. The method also showed accuracy, with average recoveries ranging from 95.25 to 108.27% and repeatability showing values between 0.14 and 7.25%. Benzo[a]pyrene was found in all chocolates, ranging from 1.58 to 4.34 µg.kg-1 of fat. Chocolate samples with 53-60% cocoa and chocolate 70% cocoa showed the high contamination for BaP; the 40% cocoa and milk chocolate samples showed low contamination. As for dietary exposure, women (by gender) and adolescents (by age) had higher DEs for BaP due to chocolate consumption, especially for 70% cocoa chocolate. The results showed low contamination by PAHs in the analyzed chocolates, however, the increased consumption by different age groups can contribute to DEs to BaP. Conclusions - The standardized methodology was considered selective and sensitive, ensuring the efficiency of the test. The analyzed samples showed results below the limits established by the European Community Regulation No. 835/2011 for the BaP and for the sum of the 4 PAHs. In view of the groups analyzed (gender, age and place of residence), chocolates 70% cocoa contributed to the greater dietary exposure to BaP. The presence of the four priority PAHs in the analyzed chocolates demonstrated the importance of programs to monitor the production chain in cocoa products (cocoa mass, cocoa butter and cocoa powder) and ingredients used in chocolate formulation (sugar and powdered milk).