This study aimed to investigate a miniaturized extraction methodology with analysis by GC-MS for simultaneous detection and quantification of cocaine (COC), benzoylecgonine (BZE) and cocaethylene (CE) in hair to monitor drug addicts. The process was done following the international guide of the Scientific Task Force on Forensic Toxicology. After sample extraction, derivatizing solution was added, clean-up was done by the Hollow Fiber Liquid Phase Microextraction (HF-LPME) and adapted to use the ultrasonic bath, a simple and easy-to-handle device. Isobutylchloroformate was first used as derivatization reagent to convert benzoylecgonine to isobutylbenzoylecgonine. Analytes quantification occurred within a linear range of 0.1 to 20 ng mg-1 for COC and 0.05 to 5 ng mg-1 for BZE and CE, with a correlation coefficient of r > 0.99. Limits of detection were 0.05; 0.03 and 0.03 ng mg-1, whereas limits of quantification were 0.1; 0.05 and 0.05 ng mg-1 for COC; BZE and CE, respectively. There was no matrix effect interference. Intra and inter-day precision and accuracy were acceptable according to international guidelines. The analytical method HF-LPME-GC-MS was successfully applied to 14 hair samples from patients admitted in drugs detoxification programs. All samples resulted positive for cocaine (0.80-> 20 ng mg-1) and benzoylecgonine (0.20-> 5 ng mg-1), 11 samples were positive for cocaethylene (0.10-> 0.60 ng mg-1).

Abstract The present study describes a method for simultaneous analysis of cocaine, benzoylecgonine and cocaethylene in urine samples. After solid phase extraction, Gas Chromatography-Mass Spectrometry was used for identification and quantification. The calibration curves were linear at 20 - 3000 ng/mL, r2 0.9997 for benzoylecgonine, 15 - 2000 ng/mL, r2 0.9985 and r2 0.9993 for cocaine and cocaethylene, respectively. Accuracy values: cocaine: 93,5 - 102,1%; benzoylecgonine: 97,5 - 104,8%; cocaethylene: 90,6 - 101,5%. Precision values: cocaine: C.V 5,4 - 14,6%; benzoylecgonine: C.V 7,8 - 12,3%; cocaethylene: C.V 5,9 - 12,3%. Detection and quantification limit values: cocaine and cocaethylene:10 ng/mL and 15 ng/mL, respectivalty; benzoylecgonine:15 ng/mL and 20 ng/mL, respectivaley. Recovery values: cocaine: 78,0 - 85,8%; benzoylecgonine: 74,0 - 79,8%; cocaethylene: 83,0 - 91,5%. The method described is advantageous compared to others, as it simultaneously detects the major analytes found in urine samples due to cocaine use and has been successfully validated.

RESUMO: Os óleos essenciais são possíveis alternativas ao uso de praguicidas sintéticos no controle da contaminação por fungos. O óleo essencial de gengibre (Zingiber officinale - OEG) é conhecido por possuir propriedades antifúngicas e antiaflatoxigênicas, mas seu uso como fumigante in situ ainda não foi estudado. O objetivo deste estudo foi avaliar os efeitos do OEG sobre Aspergillus flavus como agente fumigante em grãos de milho armazenados. Os principais compostos encontrados no OEG foram α-zingibereno (23,85%) e geranial (14,16%), caracterizados por cromatografia gasosa acoplada a espectrometria de massas e ressonância magnética nuclear. O OEG foi utilizado como fumigante em grãos de milho irradiados em concentrações variando de 5 a 50 µg/g e os efeitos resultantes foram comparados a um agente antifúngico sintético (carbendazim e tiram), antifúngico tradicionalmente usado para o tratamento de sementes. A eficácia antifúngica do OEG contra A. flavus foi comprovada de maneira dependente da dose através do teste in situ (grãos de milho). O OEG inibiu a produção de aflatoxina nas concentrações 25 e 50 µg/g e controlou o crescimento de fungos. Portanto, o OEG pode ser usado como uma alternativa eficaz e não tóxica aos tratamentos convencionais em grãos de milho armazenados para o controle natural de A. flavus.

ABSTRACT: Essential oils are possible alternatives to the use of synthetic pesticides for control of fungal contamination. Ginger (Zingiber officinale) essential oil (GEO) is known for having antifungal and antiaflatoxigenic properties, but its use as a fumigant in situ has not been studied yet. The aim of this study was to evaluate GEO’s effects upon Aspergillus flavus as a fumigant agent in stored maize grains. The main compounds reported in GEO were α-zingiberene (23.85%) and geranial (14.16%), characterized by gas chromatography-mass spectrometry and nuclear magnetic resonance. The GEO was used as a fumigant in irradiated maize grains in concentrations ranging from 5 to 50 µg/g and the resulting effects were compared to a synthetic antifungal agent (carbendazim and thiram), an antifungal traditionally used for seed treatment. The antifungal efficacy of GEO against A. flavus has been proven in a dose-dependent manner through in situ (maize grains) test. The GEO inhibited aflatoxin production at concentrations 25 and 50 µg/g and controlled fungal growth. Therefore, GEO can be used as an effective and non-toxic alternative to conventional treatments in stored maize grains for the natural control of A. flavus.

Realizou-se um estudo transversal controlado, com hortifruticultores, durante o período de outubro de 2009 a outubro de 2010, a fim de caracterizar o uso de praguicidas, estabelecer o perfil sócio demográfico e analisar a atividade das colinesterases em trabalhadores rurais do sul do Brasil. Os dados foram obtidos de 173 trabalhadores e 179 controles. Um formulário estruturado foi aplicado obtendo informações sócio demográficas, conhecimento e práticas de trabalho relacionadas com o uso de praguicidas. Os valores de referência para colinesterases totais (ChEs) e butirilcolinesterase (BuChE) foram obtidos a partir da média da atividade enzimática do grupo controle. A idade média da população exposta foi de 40,0±11,2 anos. Os sinais/sintomas foram significativamente diferentes (p<0,05) entre a população exposta e não exposta. Quarenta trabalhadores (23,1%) apresentaram sinais/sintomas relacionados com a exposição aos praguicidas. O valor médio do grupo controle foi de 6,3 μmol/mL/min para o gênero masculino e 5,6 μmol/mL/min para o gênero feminino na ChEs; 2,4 μmol/mL/min para o gênero masculino e 2,0 μmol/mL/min para o gênero feminino na BuChE. Do total de trabalhadores (n=173), 08 (4,6%) apresentaram inibição elevada (>30%) das atividades das ChEs, e para BuChE nenhum trabalhador apresentou alta inibição (>50%). Dentre os fatores que poderiam estar envolvidos destacam-se o gênero, escolaridade, orientação para trabalhar com praguicidas, exposição e medidas de higiene.

A controlled cross-sectional study of family growers of fruit and vegetables was conducted between October 2009 and October 2010 to characterize the use of pesticides, establish the socio-demographic profile, and analyze cholinesterase activity in small-scale agricultural workers in Southern Brazil. Data was collected for 173 workers and 179 controls. A structured questionnaire was applied collecting socio-demographic information and determining knowledge and work practices in relation to pesticide use. The benchmarks for total cholinesterase (ChEs) and butyrylcholinesterase (BuChE) were obtained from the average enzymatic activity of the occupationally unexposed group (control). The mean age of the exposed population was 40.0 ± 11.2 years. The symptoms differed significantly (p<0.05) between the exposed and unexposed populations. Forty (23.1%) workers showed symptoms associated with exposure to pesticides. The average value of enzymatic activity in the occupationally unexposed group for ChEs was 6.3 μmol/mL/min among males and 5.6 μmol/mL/min among females while for BuChE was 2.4 μmol/mL/min among males and 2.0 μmol/mL/min among females. Eight (4.6%) workers had high inhibition (>30%) of ChEs activity, whereas no workers showed high inhibition (>50%) of BuChE. Potential factors involved include gender, education, pesticide orientation, exposure, and hygiene measures.

Aflatoxinas são metabólitos carcinogênicos produzidos pelo Aspergillus flavus, A. parasiticus e A. nomius. No presente estudo, amostras de amendoim foram coletadas em diferentes estágios fenológicos da planta durante as safras de 2007/2008 e 2008/2009, bem como no período de armazenamento do amendoim referente à safra de 2007/2008. A micoflora e os níveis de aflatoxinas foram determinados. Os resultados demonstraram a presença de Fusarium spp., Macrophomina spp., Trichoderma spp., Aspergillus spp. e Cladosporium spp. no período de maturação (39,8; 17,9; 8,2; 2,7 e 1,7%, respectivamente) e no armazenamento (49,8; 27,8; 12,5; 8,8 e 1,0%). As aflatoxinas não foram detectadas nas amostras coletadas. Dos 25 isolados de Aspergillus spp., 24 foram produtores de aflatoxina B1 (96%), 10 de B2 (40%), 17 de G1 (68%) e 10 de G2 (40%). O isolamento de Aspergillus spp. durante o armazenamento não foi influenciado pela temperatura, umidade relativa e atividade de água (p > 0,05). A detecção de Aspergillus spp. produtores de aflatoxinas nos diferentes estágios fenológicos: ginóforo aéreo, granação (sementes), frutos maduros (vagem) e frutos secos (colheita) demonstra a importância de boas práticas agrícolas da produção ao armazenamento do amendoim no sul do Brasil.

Aflatoxins are carcinogens produced by Aspergillus flavus, A. parasiticus and A. nomius. In the present study, peanut samples were collected at different phenological stages of the plant during the 2007/2008 and 2008/2009 seasons and from stored peanuts harvested in 2007/2008. The mycoflora and aflatoxins in the peanuts were evaluated. The results showed the presence of Fusarium spp., Macrophomina spp., Trichoderma spp., Aspergillus spp. and Cladosporium spp. during the period of peanut maturation (39.8, 17.9, 8.2, 2.7 and 1.7%, respectively) and storage (49.8, 27.8, 12.5, 8.8 and 1.0%, respectively). However, aflatoxins were not detected in the samples. Of the 25 Aspergillus spp. isolates, 24 (96%) were producers of aflatoxin B1 (96%), 10 (40%) of aflatoxin B2, 17 (68%) of aflatoxin G1, and 10 (40%) of aflatoxin G2. The isolation of Aspergillus spp. during storage was not influenced by the temperature, relative humidity or water activity (p > 0.05). The detection of aflatoxin-producing Aspergillus spp. in the samples analysed at different phenological stages, aerial gynophore, pod filling (seeds), mature fruits (pod), and dry fruits (harvest), indicates the importance of good agricultural practices from the cultivation to storage of peanuts in southern Brazil.