Amphetamine, methamphetamine, and MDMA in hair samples from a rehabilitation facility: Validation and applicability of HF-LPME-GC-MS.

INTRODUCTION: It is known that drug abuse jeopardizes economic and social development. Toxicological analyses can guide prevention and treatment strategies in rehabilitation facilities. The current greatest challenge is finding easily adaptable and less costly sensitive methods that meet the principles of green chemistry. Hair, as a biological matrix, has several advantages, and its ability to detect consumption for longer periods keeping the matrix stable and unaltered stands out. This manuscript addresses the use of a miniaturized technique in an alternative matrix, by making use of a reduced amount of solvents to quantify amphetamines, aiming to guide prevention and treatment strategies in rehabilitation facilities. METHODS: A Hollow Fiber Liquid-phase Microextraction (HF-LPME) technique for extracting amphetamines from hair samples with Gas Chromatography-Mass Spectrometry (CG-MS) was validated, adapted, and applied to ten samples from patients of a rehabilitation facility. RESULTS: The technique proved to be sensitive, accurate, precise, and not affected by interference from the biological matrix and the linear range for the analytes was 0.2 to 20 ng mg -1. The three analytes were quantified in the samples analyzed. It is worth stressing that the patients were young. CONCLUSION: The HF-LPME-GC-MS technique complied with the principles of green chemistry, and proved to be a sensitive technique, adaptable to the routine of common laboratories. Validation in the analysis phase with authentic samples, thus, showed that it can be an important tool for preventing and controlling drug addiction.

Occurrence and risk assessment of population exposed to deoxynivalenol in foods derived from wheat flour in Brazil.

Deoxynivalenol (DON) is the most important of the trichothecenes in terms of amounts and occurrence in wheat. This compound was shown to be associated with a glomerulonephropathy involving an increase of immunoglobulin A in humans. This study assessed the occurrence of DON in wheat flour and the exposure of Brazilian teenagers, adults and elderly to this mycotoxin due to intake of wheat flour-based products. DON extraction in wheat flour was carried out by solid phase extraction and the quantification was performed by ultra-high proficiency liquid chromatography with diode-array detection. A total of 77.9% of all samples were positive for DON, with concentrations ranging from 73.50 to 2794.63 µg kg-1. The intake was calculated for the average and 90th percentile of the contamination levels of DON in foods based-wheat for teenagers, adults and elderly in Brazil, and compared with the provisional maximum tolerable daily intakes (PMTDI). Females of all age groups were exposed to DON at higher levels when compared to males in regard of consumption of breads and pastas. Teenagers were the main consumers of foods derived from wheat flour, with maximum probable daily intakes of 1.28 and 1.20 µg kg-1 b.w. day-1 for females and males, respectively. This population is at an increased risk of exposure to DON due to consumption of wheat flour-based foods in Brazil.

Occurrence and exposure assessment to aflatoxins in peanuts commercialized in the northwest of Parana, Brazil / Ocorrência e avaliação da exposição as aflatoxinas em amendoim comercializado na região noroeste do Paraná, Brasil

ABSTRACT: Peanuts are a nutritious food consumed worldwide. Some species of the genus Aspergillus, producers of aflatoxins, colonize peanuts. Human exposure to aflatoxins occurs by ingesting contaminated foods. The objective of this paper was to assess the occurrence and dietary exposure of the population to aflatoxins in peanuts commercially available in Maringa, Brazil, from May 2013 to April 2017. A total of 104 peanut samples were evaluated by thin-layer chromatography and confirmed by derivatization with trifluoroacetic acid. The contamination rate detected was 24.0% in average concentration of 13.4µg/kg. Twenty positive samples (19.2%) exceeded the maximum level permitted in Brazil for the sum of four aflatoxins. Estimated probable daily intake was 1,28 µg/kg body weight/day, exceeding the Provisional Maximum Tolerable Daily Intake (0.001μg/kg body weight/day). According to the high levels of aflatoxins found in peanuts, there is need for further monitoring the presence of aflatoxins in peanuts in natura to reduce the levels of contamination.

RESUMO: O amendoim é um alimento nutritivo e muito consumido no mundo todo. Algumas espécies de fungos do gênero Aspergillus, produtores de aflatoxinas, colonizam o amendoim. A exposição humana as aflatoxinas ocorre por meio do consumo de alimentos contaminados. O objetivo deste estudo foi avaliar a ocorrência e exposição da população as aflatoxinas em amendoim comercializado no município de Maringá, Brasil, durante o período de maio de 2013 a abril de 2017. Foram avaliadas 104 amostras de amendoim por análise cromatográfica e confirmadas por derivação com ácido trifluoroacético. A frequência de contaminação foi de 24,0% em concentração média de 13,4µg/kg. Vinte amostras positivas (19,2%) apresentaram-se acima do limite máximo permitido no Brasil para a somatória das quatro aflatoxinas. A ingestão diária provável média encontrada foi de 1,28µg/kg peso corpóreo/dia, superando a ingestão diária máxima tolerável provisória (0,001µg/kg peso corpóreo/dia). De acordo com os elevados teores de aflatoxinas encontrados em amendoim, há necessidade de um maior monitoramento da presença de aflatoxinas em alimentos, tendo em vista a redução nos níveis de contaminação do amendoim in natura.

Exposure to heavy metals due to pesticide use by vineyard farmers.

OBJECTIVES: To statistically analyze serum heavy metal levels in biological samples obtained from vineyard workers from southern Brazil and check for heavy metal exposure due to pesticide use. METHODS: Serum samples were obtained from 54 farmers and 108 healthy unexposed individuals. Samples from the same farmers were obtained at three different time points over a 1-year period. Levels of lead, arsenic, nickel, zinc, manganese and copper were determined for each sample using dynamic reaction cell inductively coupled plasma mass spectrometry (DRC-ICP-MS). All results were statistically analyzed using the nonparametric Kruskal-Wallis test (followed by Dunn's post hoc test). RESULTS: Results showed that serum heavy metal levels in farmers were twofold to fourfold higher than in controls. This difference was found for all heavy metals tested and was significant (p < 0.05). Serum metal levels among the farmers also correlated with the frequency of use of pesticides at a specific time of year, which varied according to seasonal conditions influencing the need for pesticide application. CONCLUSIONS: It can be concluded that in the vineyard region assessed, farmers were more susceptible to heavy metal exposure due to pesticide use.

Oxytetracycline, tetracycline, chlortetracycline and doxycycline in pasteurised cow's milk commercialised in Brazil.

The aim of this study was to investigate the presence of tetracycline residues in pasteurised cow's milk using high-performance liquid chromatography coupled with UV/VIS detection to determine the exposure of Brazilian's population to antibiotic residues. One hundred samples collected from the State of Paraná, Brazil, were analysed. Three of these samples were contaminated at the following concentrations: 121.8 µg·kg(-1) for oxytetracycline, 93.5 µg·kg(-1) for tetracycline and 134.6 µg·kg(-1) for chlortetracycline (61.6 µg·kg(-1)) and doxycycline (73.0 µg·kg(-1)). The median tetracycline residue concentration found in the samples was 42.3 µg·kg(-1), and the estimated daily intake (EDI) was 0.05 µg Kg(-1) bw day(-1) in Brazil. These results demonstrate that the occurrence of tetracycline in Brazilian milk was low (3%) and only for 2% above the maximum residue limit, so the risk to the population from the presence of these residues in milk was low (<1% of the acceptable daily intake).

Antifungal properties and inhibitory effects upon aflatoxin production of Thymus vulgaris L. by Aspergillus flavus Link.

The antifungal and antiaflatoxigenic properties of Thymus vulgaris essential oil (TEO) were evaluated upon Aspergillus flavus "in vitro". Suspension containing 10(6) of A. flavus were cultivated with TEO in concentrations ranging from 50 to 500 µg/mL. TEO reached minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) at 250 µg/mL. Inhibition of ergosterol biosynthesis was detected at a concentration of 100 µg/mL of TEO. Morphological evaluation performed by both light microscopy and scanning electron microscopy showed that antifungal activity of TEO could be detected starting at a concentration of 50 µg/mL and the fungicide effect at a concentration of 250 µg/mL. TEO completely inhibited production of both B1 and B2 aflatoxins (AFB1 and AFB2) at a concentration of 150 µg/mL. This way, fungal biomass development and aflatoxin production were dependent on TEO concentration. Therefore, TEO was capable of controlling the growth of A. flavus and its production of aflatoxins.

Perfis antropométrico, lipídico e glicêmico em adolescentes de uma instituição filantrópica no noroeste do Paraná / Anthropometric, lipids and blood glucose profiles in adolescents of a philanthropic institution in the northwest Paraná

INTRODUÇÃO: A obesidade na adolescência é um fator preditivo da obesidade no adulto. A prevalência em crianças e adolescentes tem aumentado na maior parte dos países e tem-se traduzido em um dos mais significativos problemas nutricionais da atualidade. OBJETIVOS: Este estudo teve por objetivo verificar os perfis antropométrico, lipídico e glicêmico em adolescentes de baixo nível socioeconômico de uma instituição filantrópica no noroeste do Paraná. MATERIAL E MÉTODOS: A população estudada foi composta por 92 adolescentes, com idade entre 10 e 17 anos. Foi utilizado o índice de massa corporal (IMC) para verificar o estado nutricional dos adolescentes, considerando normal 24,9 kg/m² e sobrepeso > 25 kg/m². A razão cintura-quadril (RCQ) foi mensurada por meio do quociente da cintura pelo quadril, e as concentrações de glicose, triglicerídeos, colesterol total, frações HDL-C, LDL-C e VLDL-C foram determinadas segundo o método enzimático colorimétrico. RESULTADOS: Encontraram-se 9,79 por cento de sobrepeso nos adolescentes e 9,78 por cento de RCQ e colesterol inadequados para a idade. Baixa correlação foi encontrada entre IMC e sobrepeso, glicemia, hipercolesterolemia, hipertrigliceridemia, LDL-C, VLDL-C e HDL-C, enquanto idade (razão de chance [RC] = 49,16) e RCQ (RC = 13,99) apresentaram correlação significativa (p < 0,05) com IMC. CONCLUSÃO: O resultado demonstra a necessidade de políticas públicas de saúde no âmbito escolar como fator preventivo da obesidade e seus fatores de risco.

INTRODUCTION: Obesity in adolescence is a predictive factor of obesity in adulthood. The prevalence in children and adolescents has increased in most countries and has become one of the most significant nutritional problems nowadays. OBJECTIVES: This study aimed at verifying the anthropometric, lipid and glycemic profiles in lower class adolescents from a philanthropic institution located in the Northeast of Paraná State. MATERIAL AND METHOD: The studied population consisted of 92 adolescents, from 10 to 17 years of age. The Body Mass Index (BMI) was used to verify their nutritional status, considering < 24.9 kg/m² normal and > 25 kg/m² overweight. The waist-to-hip measurement was calculated through waist-to-hip ratio (WHR) and glucose concentrations, triglycerides, total cholesterol, HDL-c, LDL-c and VLDL-c fractions were determined through enzymatic methods. RESULTS: 9.79 percent were overweight and 9.78 percent of WHR and cholesterol levels were inadequate to the age group. BMI (overweight), blood glucose, hypercholesterolemia, hypertriglyceridemia, LDL-c, VLDL-c and HDL-c showed low correlation, whereas age (OR = 49.16) and WHR (OR = 13.99) showed significant correlation p < 0.05. CONCLUSION: The result evidenced the necessity of health public policies in educational scope in order to prevent obesity and its risks factors in adolescence.

Presynaptic M1, M2, and A1 receptors play roles in tetanic fade induced by pancuronium or cisatracurium.

PURPOSE: We investigated whether presynaptic facilitatory M1 and/or inhibitory M2 muscarinic receptors contributed to pancuronium- and cisatracurium-induced tetanic fade. METHODS: Phrenic nerve-diaphragm muscle preparations of rats were indirectly stimulated with tetanic frequency (75 +/- 3.3 Hz; mean +/- SD). Doses of pancuronium, cisatracurium, hexamethonium, and d-tubocurarine for producing approximately 25% fade were determined. The effects of pirenzepine and methoctramine, blockers of presynaptic M1 and M2 receptors, respectively, on the tetanic fade were investigated. RESULTS: The concentrations required for approximately 25% fade were 413 microM for hexamethonium (26.8 +/- 2.4% 4% fade), 55 nM for d-tubocurarine (28.7 +/- 2.55% fade), 0.32 microM for pancuronium (25.4 +/- 2.2% fade), and 0.32 microM for cisatracurium (24.7 +/- 0.8% fade). Pirenzepine or methoctramine alone did not produce the fade. Methoctramine, 1 microM, attenuated the fade induced by hexamethonium (to 16.0 +/- 2.5% fade), d-tubocurarine (to 6.0 +/- 1.6 fade), pancuronium (to 8.0 +/- 4.0% fade), and cisatracurium (to 11.0 +/- 3.3% fade). 10 nM pirenzepine attenuated only the fades produced by pancuronium (to 5.0 +/- 0.11% fade) and cisatracurium (to 13.3 +/- 5.3% fade). Cisatracurium (0.32 microM) showed antiacetylcholinesterase activity (in plasma, 14.2 +/- 1.6%; 6%; in erythrocyt 17.2 +/- 2.66%) similar to that of pancuronium (0.32 microM). The selective A1 receptor blocker, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX; 2.5 nM), also attenuated the fades induced by pancuronium and cisatracurium. CONCLUSION: The tetanic fades produced by pancuronium and cisatracurium depend on the activation of presynaptic inhibitory M2 receptors; these agents also have anticholinesterase activities. The fades induced by these agents also depend on the activation of presynaptic inhibitory A1 receptors through the activation of stimulatory M1 receptors by acetylcholine.

Intralaboratory optimization and validation of a method for patulin determination in grapes by Thin-Layer Chromatography

The aim of this work was to optimize and validate, by intralaboratorial procedures, a method for the determination of patulin in grapes by thin-layer chromatography. The steps of extraction, cleanup, detection and quantification were optimized. For the validation of the method, recovery assays with standard solutions and artificially contaminated samples were carried out. The mean recovery and the variation coefficient were 65.4 percent and 7.58 percent, respectively. The optimized conditions were: 50 mL of grape juice, three extraction stages (with 100 mL of ethyl acetate in the first stage and 50 mL in second and third stages), and 100 æL of ethanol to solubilize the extract. The solvent-system used was toluene - ethyl acetate - formic acid (6:3:1), and 0.5 percent MBTH in 5 percent formic acid was sprayed on the plates to intensify the fluorescence. The visual detection and quantification limits were 7.44 ng and 15.87 æg.kg-1, respectively. The optimized and validated method demonstrated sufficient efficiency for adoption in the monitoring of patulin in grape.

O objetivo deste trabalho foi otimizar e validar, por procedimentos intralaboratoriais, um método de determinação de patulina em uva por cromatografia em camada delgada. Foram realizados testes de otimização das etapas de extração, limpeza, detecção e quantificação. Para validação do método foram realizados ensaios de recuperação com soluções padrões e amostras artificialmente contaminadas. A recuperação e o coeficiente de variação foram 65,4 por cento e 7,58 por cento, respectivamente. As condições otimizadas foram: 50 mL do suco da uva; três etapas de extração, 100 mL de acetato de etila na primeira etapa e 50 mL na segunda e terceira etapas; 100 æL de etanol para solubilizar o extrato; a fase móvel tolueno-acetato de etila-ácido fórmico (6:3:1) e o revelador o MBTH 0,5 por cento em ácido fórmico 5 por cento. O limite de detecção visual foi de 7,44 ng e o de quantificação de 15,87 æg/kg. O método otimizado e validado apresentou eficiência para ser adotado em atividades de monitoramento de patulina em uva.

Biomarcadores para avaliação da exposição humana às micotoxinas / Biomarkers for assessment of human exposure to mycotoxins

Atualmente, as micotoxinas representam um risco de contaminação ambiental, acarretando sérios prejuízos à saúde humana. Essas toxinas podem estar presentes em diferentes tipos de alimentos, que constituem a principal fonte de exposição para o homem. As exposições podem ser monitoradas através do uso de biomarcadores, que elucidam a relação causa/efeito e dose/efeito na avaliação de risco à saúde para fins de diagnóstico clínico e laboratorial. Realizou-se uma revisão bibliográfica do período de 1981-2005, no MEDLINE, sobre utilização e propostas de biomarcadores para a exposição a aflatoxinas, fumonisinas, desoxinivalenol e ocratoxina A. Os possíveis biomarcadores para avaliar a exposição humana às aflatoxinas foram os metabólitos urinários de aflatoxina B1, como aflatoxina M1, aflatoxina P1, aflatoxina Q1, aflatoxina livre em soro ou plasma, os adutos de AFB-N7-guanina, os adutos de albumina ou mutação no gene supressor de tumor p53, presentes em fluidos biológicos. Para as fumonisinas, os biomarcadores foram os níveis de fumonisina B1 e fumonisina B2 livres, ou de esfinganina e esfingosina em sangue e urina. O desoxinivalenol tem como biomarcadores de exposição os produtos de seu metabolismo e adutos macromoleculares (proteína/DNA) presentes nos fluidos biológicos. Para a exposição à ocratoxina A (OA) os biomarcadores se restringem à quantificação da própria toxina nos fluidos biológicos. A avaliação da exposição às micotoxinas constitui um importante aspecto para a saúde pública, tendo em vista a possibilidade de prevenir ou minimizar a incidência de doenças decorrentes da sua interação com o organismo.

Currently, mycotoxins represent a risk of environmental contamination, causing serious damages to human health. Those toxins can be found in different kinds of foods, and they constitute the main source of human exposure. The evaluation of such exposures can be monitored through the use of biomarkers, which elucidates the cause/effect and dose/effect relation in the evaluation of health risks for clinical and laboratory diagnostic purposes. The MEDLINE review about the use of biomarkers for assessment of aflatoxins, fumonisins, deoxynivalenol and ochratoxin A was carried out from 1981 to 2005. The biomarkers for assessment of human exposure to aflatoxins were the urinary metabolites of aflatoxin B1: aflatoxin M1, aflatoxin P1, aflatoxin Q1, the free aflatoxin in serum or plasma, the AFB-N7-guanine adducts and the albumin adducts or mutation in the tumour suppressor gene p53 present in human biological fluids. As far as fumonisins are concerned, levels of free fumonisin B1 and fumonisin B2, or levels of sphinganine and sphingosin, were quantified in blood and urine. As exposure biomarkers, deoxynivalenol has its own metabolism products and adducts (protein/DNA) present in human fluids. As to ochratoxin A exposure, we measure it in biological fluids, once it enables us to prevent or minimize the incidence of deaths or illnesses provoked by chemical exposure.

Ocorrência de cloranfenicol em leite pasteurizado comercializado no Estado do Paraná, Brasil / Occurence of chloramphenicol in pasteurized milk commercialized in the state of Paraná

A presença de resíduos de cloranfenicol no leite pode causar efeitos tóxicos na população exposta. O objetivo deste trabalho foi investigar a ocorrência de cloranfenicol em amostras de leite comercializadas no Estado do Paraná, Brasil. Foram analisadas 151 amostras de leite pasteurizado B e C de 61 marcas diferentes, comercializadas em 35 municípios. Os resíduos de cloranfenicol foram analisados por ensaio imunoenzimático (ELISA). Foram detectados resíduos de cloranfenicol em quatro (2,6%) das amostras analisadas, sendo duas amostras de leite tipo B e duas do tipo C. Esses dados demonstram que esse antimicrobiano continua sendo utilizado em animais destinados ao consumo humano ou há presença de contaminantes ambientais. Portanto, há necessidade de programas de monitoramento de resíduos de medicamentos veterinários em alimentos para garantir sua qualidade e, conseqüentemente, proteção à saúde humana.A presença de resíduos de cloranfenicol no leite pode causar efeitos tóxicos na população exposta. O objetivo deste trabalho foi investigar a ocorrência de cloranfenicol em amostras de leite comercializadas no Estado do Paraná, Brasil. Foram analisadas 151 amostras de leite pasteurizado B e C de 61 marcas diferentes, comercializadas em 35 municípios. Os resíduos de cloranfenicol foram analisados por ensaio imunoenzimático (ELISA). Foram detectados resíduos de cloranfenicol em quatro (2,6%) das amostras analisadas, sendo duas amostras de leite tipo B e duas do tipo C. Esses dados demonstram que esse antimicrobiano continua sendo utilizado em animais destinados ao consumo humano ou há presença de contaminantes ambientais. Portanto, há necessidade de programas de monitoramento de resíduos de medicamentos veterinários em alimentos para garantir sua qualidade e, conseqüentemente, proteção à saúde humana.

The presence of chloramphenicol residues in milk can cause toxic effects on the exposed population. The aim of this work was to investigate the occurrence of chloramphenicol in milk samples commercialized in the state of Paraná, Brazil. 151 pasteurized milk samples, type B and C of 61 different marks, commercialized in 35 cities were analyzed. Chloramphenicol residues were examined by enzyme immunoassay (ELISA). Chloramphenicol residues were detected in four (2.6%) out of the milk samples analyzed, being two samples of type B and two of type C. These data show that either this antibiotic is still being used in animals destined to human consumption or there is environmental contamination. Thus, monitoring programs are needed to verify veterinary drug residues in food to certify its quality and, consequently, protection of human health.

Monitorização terapêutica de ciclosporina em um grupo de pacientes transplantados reanis / Blood monitoring of cyclosporine in renal transplant patiens

A ciclosporina é um dos imunossupressores mais utilizados em transplantes. neste trabalho foram avaliados resultados das dosagens sangíneas de ciclosporina de pacientes transplantados renais atendidos pelo Laboratório de Toxicologia da Universidade Estadual de Maringá (UEM) em 2001. A monitorização compreendru dosagens em C0 (concentração anterior à ingestão da próxima dose) e C3 (concentração três horas após a ingestão do medicamento), por imunofluorescência polarizada. Infromações sobre sexo, idade, data de transplante e medicamentos utilizados, foram obtidas. 82% dos pacientes tinham entre 25-59 anos, com predominância do sexo masculino (68%). Cicliosporina-Neoral®, azatioprina e prednisona constituiu o esquema terapêutico mais utilizado (72%). em C0, 83,6% dos resultados apresentaram-se dentro, enquanto que em C3,62,4% mostraram-se acima da faixa terapêutica recomendada (100 a 400 ng/ml). Não houve relação entre C0 ou C3 e tempo de transplante ou tratamento imunossupressor. A monitorização terapêutica é fundamental para minimizar efeitos tóxicos ou mesmo a rejeição do rrgão transplantado...

Antibacterial activity of Ocimum gratissimum L. essential oil

The essential oil (EO) of Ocimum gratissimum inhibited Staphylococcus aureus at a concentration of 0.75 mg/ml. The minimal inhibitory concentrations (MICs) for Shigella flexineri, Salmonella enteritidis, Escherichia coli, Klebsiella sp., and Proteus mirabilis were at concentrations ranging from 3 to 12mg/ml. The endpoint was not reached for Pseudomonas aeruginosa (>=24 mg/ml). The MICs of the reference drugs used in this study were similar to those presented in other reports. The minimum bactericidal concentration of EO was within a twofold dilution of the MIC for this organism. The compound that showed antibacterial activity in the EO of O. gratissimum was identified as eugenol and structural findings were further supported by gas chromatography/mass spectra retention time data. The structure was supported by spectroscopic methods.