Determination and Risk Assessment of Six Hepatotoxic Pyrrolizidine Alkaloids in Verbenae Herba / 中国实验方剂学杂志

Objective:To establish ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for simultaneous determination of six hepatotoxic pyrrolizidine alkaloids in Verbenae Herba， and to carry out preliminary risk assessment according to the research results. Method:An ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 μm） was used for analysis with 0.05% formic acid and 2.5 mmol·L^-1 ammonium formate in water （A）-0.05% formic acid and 2.5 mmol·L^-1 ammonium formate in acetonitrile （B） as mobile phase for gradient elution （0-12 min， 3%-8%B； 12-25 min， 8%-15%B； 25-26 min， 15%-3%B； 26-30 min， 3%B）， the flow rate was 0.3 mL·min^-1， the column temperature was 40 ℃， and the injection volume was 1 μL. MS system was operated by electrospray ionization （ESI） in the positive ion mode with multiple reaction monitoring mode. MS parameters of triple quadrupole and six analytes were optimized for qualitative and quantitative analysis. According to the determination results， the risk assessment was carried out by using margin of exposure （MOE） combined with transfer rate of hot water extraction. Result:Based on the instrument precision， linear range， repeatability， stability， recovery and other methodological validations， the results were in conformity with relevant standards of quantitative analysis. The linear ranges of intermedine， lycopsamine， intermedine <italic>N</italic>-oxide， lycopsamine<italic> N</italic>-oxide， echimidine<italic> N</italic>-oxide and echimidine were good （<italic>r</italic>≥0.999 0） between peak area and mass concentration in the ranges of 0.984-49.20， 0.994-49.70， 1.012-50.60， 1.032-51.60， 1.004-50.20， 1.016-50.80 µg·L^-1， respectively. The average recoveries of these six analytes were 87.2%-94.2% with relative standard deviation （RSD）<4.0%. Their MOE values were >10 000. Conclusion:The UPLC-MS/MS established in this study is stable and feasible， which can provide scientific basis for the quality control and safety evaluation of hepatotoxic pyrrolizidine alkaloids in Verbenae Herba.

Estimate of dietary intake of chloropropanols (3-MCPD and 1, 3-DCP) and health risk assessment / Estimativa da ingestão de cloropropanóis (3-MCPD e 1,3-DCP) e avaliação do risco à saúde

Chloropropanols, including 3-monochloropropane-1,2-diol (3-MCPD) and 1,3-dichloropropan-2-ol (1,3-DCP), comprise a group of chemicalcontaminants with carcinogenic and genotoxic properties. They have been found in a variety of processed foods and food ingredients, suchas hydrolyzed vegetable protein, soy sauce, cereal-based products, malt-derived ingredients, and smoked foods. This study aimed to assessthe dietary exposure to 3-MCPD and 1,3-DCP in Brazil and verify whether the presence of these substances in foods could represent healthrisks. The intake was calculated by combining data on food consumption, provided by the Consumer Expenditure Survey 2008-2009, withthe levels of contaminant occurrence determined by gas chromatography-mass spectrometry. The exposure to 3-MCPD ranged from 0.06 to0.51 µg.kg bw–1.day–1 considering average and high consumers, while the intake of 1,3-DCP was estimated to be 0.0036 µg.kg bw–1.day–1 inthe worst case scenario evaluated. Based on these results, it was verified that the Brazilians’ exposure to chloropropanols does not present asignificant health risk. However, the consumption of specific foods containing high levels of 3-MCPD could exceed the provisional maximumtolerable daily intake of 2 μg.kg bw–1 established for this compound and, therefore, represent a potential concern.

Os cloropropanóis, entre eles o 3-monocloropropano-1,2-diol (3-MCPD) e o 1,3-dicloropropan-2-ol (1,3-DCP), compreendem um grupo de contaminantes químicos com propriedades carcinogênicas e genotóxicas, encontrados em diversos alimentos processados e ingredientes alimentícios, como proteína vegetal hidrolisada, molho de soja, produtos à base de cereais, ingredientes derivados de malte e alimentos defumados. Este trabalho teve como objetivo avaliar a exposição ao 3-MCPD e 1,3-DCP pela dieta no Brasil e verificar se a presença destes compostos em alimentos pode representar um risco à saúde da população. A ingestão foi calculada combinando-se dados sobre o consumo alimentar, fornecidos pela Pesquisa de Orçamento Familiares 2008-2009, com os níveis de ocorrência dos contaminantes, determinados por cromatografia gasosa – espectrometria de massas. A exposição ao 3-MCPD variou de 0,06 a 0,51 µg.kg pc–1.dia–1 considerando médios e grandes consumidores, enquanto que a ingestão de 1,3-DCP foi estimada em 0,0036 µg.kg pc–1.dia–1 no pior cenário avaliado. Com base nesses resultados, verificou-se que a exposição aos cloropropanóis não representa um risco significativo à saúde da população brasileira.Entretanto, o consumo de determinados alimentos contendo altos níveis de 3-MCPD poderia ultrapassar a ingestão diária máxima tolerável provisória de 2 μg.kg pc–1 estabelecida para este composto e, assim, representar uma preocupação potencial.