Eloisa Dutra Caldas

Simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquid–liquid extraction with low temperature purification (LLE–LTP)

The aims of this study were to optimize and validate a methodology for the simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 (AFB1, AFB2, AFG1, AFG2, AFM1) and ochratoxin A (OTA) in breast milk, and to analyze these mycotoxins in samples obtained from human milk banks in the Federal District, Brazil. The optimized analytical method was based on liquid-liquid extraction with low temperature purification (3.25mL of acidified acetonitrile+0.75mL of ethyl acetate), followed by analysis by high-performance liquid chromatography with fluorescence detector (HPLC/FLD) and a photochemical post-column reactor. Limits of quantification (LOQ) ranged from 0.005 to 0.03ng/mL, recoveries from 73 to 99.5%, and relative standard deviations (RSD) from 1.8 to 17.3%. The LLE-LTP extraction method was shown to be simple and cost-effective, since no columns were needed for clean-up. Only 2 of the 224 breast milk samples analyzed were positive for the mycotoxins, both samples containing AFB2 at the LOQ level (0.005ng/mL). The identity of the mycotoxin detected was confirmed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This result indicates that infants who are fed with breast milk from the milk banks are not at risk from aflatoxin and ochratoxin exposure.

Avaliação de risco crônico da ingestão de resíduos de pesticidas na dieta brasileira

OBJECTIVE To conduct a chronic dietary risk assessment of the pesticides registered in Brazil up until 1999. METHODS The Theoretical Maximum Daily Intake (TMDI) for each pesticide was calculated using the Brazilian maximum residue limits and food consumption data from IBGE, the Brazilian Statistical Institute. The risk characterization was done comparing the TMDI with the acceptable daily intakes (ADI) from other countries and from the Codex Alimentarius. RESULTS The TMDI was higher than the ADI (%ADI>100) at least in one Brazilian metropolitan region for 23 pesticides. Sixteen compounds are organophosphate insecticides, with methyl parathion having the TMDI exceeding the most toxicological parameter (%ADI N=9,300). Rice, beans, citrus and tomato were the commodities which most contributed to the ingestion. From the compounds under higher risk, only 6 were registered according to the Law 98.816/90, which concerns the use of pesticides in the country. CONCLUSIONS The compounds identified in the study as presenting a potential health concern to the Brazilian consumers, and the commodities which most contributed to the ingestion, should be prioritized by the government in pesticide residue monitoring programs and in the re-registration process. In addition, residue data in food as consumed, processing factors and appropriate consumption data should be generated to allow further studies.OBJECTIVE: To conduct a chronic dietary risk assessment of the pesticides registered in Brazil up until 1999. METHODS: The Theoretical Maximum Daily Intake (TMDI) for each pesticide was calculated using the Brazilian maximum residue limits and food consumption data from IBGE, the Brazilian Statistical Institute. The risk characterization was done comparing the TMDI with the acceptable daily intakes (ADI) from other countries and from the Codex Alimentarius. RESULTS: The TMDI was higher than the ADI (%ADI>100) at least in one Brazilian metropolitan region for 23 pesticides. Sixteen compounds are organophosphate insecticides, with methyl parathion having the TMDI exceeding the most toxicological parameter (%ADIN=9,300). Rice, beans, citrus and tomato were the commodities which most contributed to the ingestion. From the compounds under higher risk, only 6 were registered according to the Law 98.816/90, which concerns the use of pesticides in the country. CONCLUSIONS: The compounds identified in the study as presenting a potential health concern to the Brazilian consumers, and the commodities which most contributed to the ingestion, should be prioritized by the government in pesticide residue monitoring programs and in the re-registration process. In addition, residue data in food as consumed, processing factors and appropriate consumption data should be generated to allow further studies.OBJECTIVE: To conduct a chronic dietary risk assessment of the pesticides registered in Brazil up until 1999. METHODS: The Theoretical Maximum Daily Intake (TMDI) for each pesticide was calculated using the Brazilian maximum residue limits and food consumption data from IBGE, the Brazilian Statistical Institute. The risk characterization was done comparing the TMDI with the acceptable daily intakes (ADI) from other countries and from the Codex Alimentarius. RESULTS: The TMDI was higher than the ADI (%ADI>100) at least in one Brazilian metropolitan region for 23 pesticides. Sixteen compounds are organophosphate insecticides, with methyl parathion having the TMDI exceeding the most toxicological parameter (%ADIN=9,300). Rice, beans, citrus and tomato were the commodities which most contributed to the ingestion. From the compounds under higher risk, only 6 were registered according to the Law 98.816/90, which concerns the use of pesticides in the country. CONCLUSIONS: The compounds identified in the study as presenting a potential health concern to the Brazilian consumers, and the commodities which most contributed to the ingestion, should be prioritized by the government in pesticide residue monitoring programs and in the re-registration process. In addition, residue data in food as consumed, processing factors and appropriate consumption data should be generated to allow further studies.

Susceptibility of Aedes aegypti larvae to the insecticide temephos in the Federal District, Brazil

OBJECTIVE To study the susceptibility status of Aedes aegypti to the organophosphate insecticide temephos. METHODS Samples of Ae. aegypti larvae were obtained, using ovitraps, from eight cities of the Federal District, central Brazil, in 2000 and 2001. Larvae were submitted to the diagnostic dose of 0.012 mg/l temephos, as recommended by standard World Health Organization methodology. Field populations were tested in parallel with reference strains Rockefeller and DIVAL, from the Environmental Surveillance Directory (DIVAL) insectary. The concentration and purity of temephos solutions were verified by gas chromatography. Correlation calculations were performed using StatView - SAS Institute Inc., version 5. Students t test was used for detecting differences in susceptibility, with significance levels of alpha=0.05. RESULTS In 2000, Ae. aegypti larvae populations from Taguatinga, Guara, and Nucleo Bandeirante showed resistance to temephos, with mortality ranging from 54.1 to 63.4%. The populations from Gama, Planaltina, and Sobradinho showed altered levels of susceptibility (mortality ranging from 83.6 to 92.8%). The population from Ceilandia was the only susceptible one, with 98% mortality. In 2001, all populations tested were resistant (44.4 to 66.4% mortality). No significant correlation was found between the susceptibility of populations and the distance between the cities of origin, or the amount of insecticide applied in the years preceding the study. CONCLUSIONS Ae. aegypti susceptibility to temephos is changing in the Federal District. It is essential to continue monitoring the resistance of this vector to insecticides in order to ensure the efficiency of programs aimed at vector control and at the protection of human health.

Percepção de risco, atitudes e práticas no uso de agrotóxicos entre agricultores de Culturama, MS

OBJETIVO: Avaliar a percepcao de risco, praticas e atitudes no uso de agrotoxicos por agricultores. PROCEDIMENTOS METODOLOGICOS: Estudo qualitativo realizado em Culturama, Mato Grosso do Sul, em fevereiro de 2005. Sete grupos focais (N=40), com cinco a sete integrantes cada, discutiram questoes relacionadas a agrotoxicos, incluindo a apresentacao da embalagem de um inseticida para subsidiar discussao sobre rotulos e bulas. As falas foram gravadas, transcritas e analisadas seguindo o metodo de analise do discurso. ANALISE DOS RESULTADOS: Os agricultores se mostraram cientes dos riscos de exposicao direta e indireta ao utilizar agrotoxicos; muitos se mostraram preocupados com a contaminacao potencial do meio ambiente. As informacoes que os agricultores tinham sobre agrotoxicos eram restritas principalmente a dosagem do produto, cuja principal fonte eram os revendedores. Os agricultores reclamaram do tamanho das letras e da linguagem tecnica do rotulo e da bula, mas muitos souberam interpretar os pictogramas e o codigo de cor de toxicidade presentes neles. CONCLUSOES: Os agricultores nem sempre transformam sua percepcao de risco e suas experiencias pessoais em atitudes e praticas mais seguras no uso de agrotoxicos, como o uso adequado de equipamentos de protecao individual. Eles sentem-se indefesos diante das situacoes de risco, principalmente devido aos fatores ambientais nao controlaveis e a vulnerabilidade economica. Sao essenciais programas governamentais de extensao agricola que enfatizem tecnicas alternativas de manejo de pragas e praticas seguras de uso de agrotoxicos, direcionados a essa populacao.

Uso de agrotóxicos e suicídios no Estado do Mato Grosso do Sul, Brasil

Prevalence of suicide with pesticides in the State of Mato Grosso do Sul, Brazil, was evaluated based on data from the Integrated State Center for Toxicological Surveillance under the State Health Department and reported from January 1992 to December 2002. Population and crop production data were collected from the Brazilian Institute of Geography and Statistics, and suicide data were obtained from the State Health Department. During the period studied, 1,355 cases of pesticide poisoning were reported, including 506 suicide attempts, resulting in 139 deaths. The regions of Campo Grande and Dourados had the highest prevalence of suicide attempts, with Dourados having the most deaths. Dourados also had a high prevalence of suicide attempts overall, with an increasing trend in the previous 10 years. The results indicated that Dourados is a critical region in the State in terms of intentional ingestion of pesticides, showing the need for an epidemiological investigation to better evaluate and quantify these events among the rural population.

Arsenic, lead, mercury and cadmium: Toxicity, levels in breast milk and the risks for breastfed infants.

Metals are ubiquitous in nature, being found in all environmental compartments, and have a variety of applications in human activities. Metals are transferred by maternal blood to the fetus via the placenta, and exposure continues throughout life. For the general population, exposure comes mainly from water and food consumption, including breast milk. In this paper, we reviewed studies on the toxicity of arsenic, lead, mercury and cadmium, the toxic metals of most concern to human health, focusing on the potential risks to newborns and infants. A total of 75 studies published since 2000 reporting the levels of these metals in breast milk were reviewed. Lead was the metal most investigated in breast milk (43 studies), and for which the highest levels were reported (up to 1515µg/L). Arsenic was the least investigated (18 studies), with higher levels reported for breast milk (up to 149µg/L) collected in regions with high arsenic concentrations in water (>10µg/L). Data from 34 studies on mercury showed that levels in breast milk were generally higher in populations with high fish consumption, where it may be present mainly as MeHg. Cadmium levels in breast milk were the lowest, with means <2µg/L in most of the 29 studies reviewed. Results of risk assessments indicated that the intake of arsenic, lead and mercury by infants through breastfeeding can be considered a health concern in most regions of the world. Although the potential risks to infants are mostly outweighed by the benefits of breast milk consumption, it is essential that contaminants be continuously monitored, especially in the most critical regions, and that measures be implemented by health authorities to reduce exposure of newborns and infants to these metals, and thus avoid unnecessary health risks.

Prescription and illicit psychoactive drugs in oral fluid—LC–MS/MS method development and analysis of samples from Brazilian drivers

This study is part of a larger project designed to investigate the prevalence of psychoactive drug (PAD) use among Brazilian drivers. In this paper we describe the development and validation of an analytical method to analyze 32 prescription and illicit PADs (amphetamines, benzodiazepines, cocaine, cannabis, opioids, ketamine and m-CPP) and metabolites in oral fluid samples collected with a Quantisal™ device. Samples were extracted with ethyl acetate:hexane and analyzed by LC-MS/MS. Instrumental LOD ranged from 0.26 to 0.65 ng/mL. Mean procedural recoveries at 1.3 ng/mL (LLOQ) ranged from 50% to 120% for 24 compounds. Recoveries were concentration independent, with the exception of femproporex, heroin and ecgonine methyl-ester (EME) for which the recovery decreased significantly at higher levels (13 and 52 ng/mL). RSD was <20% for all compounds at all spiking levels. Ion suppression due to the matrix was <20% for most compounds, and higher than 60% for EME and diethylpropion. Analysis was performed against a in-matrix standard curve. About 10% of the 2235 oral fluid samples collected from drivers on Brazilian Federal highways were positive (≥LOD) for at least one analyte investigated. Alone or in combination with other drugs, cocaine/metabolites were the analytes most detected in the samples (129; 5.8%), followed by amphetamines/metabolite (69; 3.1%), benzodiazepines (28; 1.2%), cannabinoids (23; 1.1%) and opioids (8; 0.4%). Detection of at least two PADs from different classes accounted for 9.3% of the 236 positive samples. Cocaine was found at higher levels in the samples (up to 1165 ng/mL). Preventive measures aimed at reducing the use of PADs by drivers in Brazil will certainly contribute to decrease the countrys highway death rates.

Pesticide residues in cashew apple, guava, kaki and peach: GC-μECD, GC-FPD and LC-MS/MS multiresidue method validation, analysis and cumulative acute risk assessment.

A multiresidue method for the determination of 46 pesticides in fruits was validated. Samples were extracted with acidified ethyl acetate, MgSO4 and CH3COONa and cleaned up by dispersive SPE with PSA. The compounds were analysed by GC-FPD, GC-μECD or LC-MS/MS, with LOQs from 1 to 8 μg/kg. The method was used to analyse 238 kaki, cashew apple, guava, and peach fruit and pulp samples, which were also analysed for dithiocarbamates (DTCs) using a spectrophotometric method. Over 70% of the samples were positive, with DTC present in 46.5%, λ-cyhalothrin in 37.1%, and omethoate in 21.8% of the positive samples. GC-MS/MS confirmed the identities of the compounds detected by GC. None of the pesticides found in kaki, cashew apple and guava was authorised for these crops in Brazil. The risk assessment has shown that the cumulative acute intake of organophosphorus or pyrethroid compounds from the consumption of these fruits is unlikely to pose a health risk to consumers.

Behavioural and neurotoxic effects of ayahuasca infusion (Banisteriopsis caapi and Psychotria viridis) in female Wistar rat.

Ayahuasca, a psychoactive beverage used by indigenous and religious groups, is generally prepared by the coction of Psychotria viridis and Banisteriopsis caapi plants containing N,N-dimethyltryptamine (DMT) and β-carboline alkaloids, respectively. To investigate the acute toxicity of ayahuasca, the infusion was administered by gavage to female Wistar rats at doses of 30X and 50X the dose taken during a religious ritual, and the animals observed for 14 days. Behavioural functions were investigated one hour after dosing at 15X and 30X using the open field, elevated plus maze, and forced swimming tests. Neuronal activation (c-fos marked neurons) and toxicity (Fluoro-Jade B and Nissl/Cresyl staining) were investigated in the dorsal raphe nuclei (DRN), amygdaloid nucleus, and hippocampal formation brain areas of rats treated with a 30X ayahuasca dose. The actual lethal oral dose in female Wistar rats could not be determined in this study, but was shown to be higher than the 50X (which corresponds to 15.1mg/kg bw DMT). The ayahuasca and fluoxetine treated groups showed a significant decrease in locomotion in the open field and elevated plus-maze tests compared to controls. In the forced swimming test, ayahuasca treated animals swam more than controls, a behaviour that was not significant in the fluoxetine group. Treated animals showed higher neuronal activation in all brain areas involved in serotoninergic neurotransmission. Although this led to some brain injury, no permanent damage was detected. These results suggest that ayahuasca has antidepressant properties in Wistar female at high doses, an effect that should be further investigated.

Aflatoxins in food products consumed in Brazil: a preliminary dietary risk assessment

A preliminary dietary exposure assessment for aflatoxins (AFs; AFB1, AFB2, AFG1 and AFG2) was conducted to evaluate the potential carcinogenic risks for the Brazilian population. AF concentration data in food were obtained from analysis reports issued by the Central Public Health Laboratory of the Federal District (LACEN-DF) and from published work. Food consumption and body weight (bw) data were obtained from a national survey conducted in 2008/2009. Cancer risks arising from exposure to aflatoxins were assessed using the carcinogenic potency of AFs estimated by the JECFA, and hepatitis B virus prevalence in the Brazilian population. Additionally, margins of exposure (MOE) were also calculated for the various scenarios investigated. A total of 942 food samples were analysed for AFs in the Federal District between 2002 and 2011 with 4.5% of them being positive for at least one aflatoxin (LOQ = 2 µg kg−1). The highest percentage of contamination was found in peanuts (8.1%) and Brazil nuts (6.0%), with mean levels ranging from 6.7 µg kg−1 in peanut products to 36.9 µg kg−1 in Brazil nuts. Most of the studies conducted elsewhere in Brazil found similar results. Total AF intake for the total Brazilian population and high consumers of food relevant for AF contamination in Brazil (upper bound; samples < LOQ = 0.5 LOQ) were 6.8 and 27.6 ng kg−1 bw day−1, respectively. Cancer risk reached 0.0753 cancers year−1 per 105 individuals for the total population and 0.3056 cancers year−1 per 105 individuals for high consumers. MOE reached 25 and 6 for the total population and high consumers, respectively, indicating a potential risk for consumers. Aflatoxins are genotoxic carcinogens, and government action should be maintained and continuously improved in order to guarantee that human exposure levels are kept as low as possible.