Cocaína, metabólitos e adulterantes no esgoto : estratégias para estimar o poliuso de drogas e o perfil de usuários

Dissertação (mestrado)—Universidade de Brasília, Instituto de Química, Programa de Pós-Graduação em Química, 2018.A análise do esgoto gerado por uma população tem fornecido informações complementares ao monitoramento do consumo de drogas. Esta ferramenta, conhecida como epidemiologia do esgoto, vem sendo empregada no Distrito Federal desde 2010 para estimar o consumo de cocaína. Dados anteriores mostraram que o consumo de cocaína é maior durante fins de semana, sugerindo que podem existir grupos distintos de usuários. Um dos grupos constituído por dependentes químicos que fazem uso constante da droga e outro formado por pessoas que fazem uso intermitente da droga e podem dobrar o consumo de cocaína no fim de semana. Para tentar diferenciar situações como esta, uma alternativa seria monitorar o cocaetileno (COE), um produto de metabolização da cocaína formado na presença de etanol, que pode indicar o consumo concomitante da droga com álcool. Neste contexto, o presente trabalho buscou quantificar cocaína, bem como seus metabólitos benzoilecgonina (BE) e cocaetileno, além dos adulterantes, fenacetina (FEN) e levamisol (LEV) em amostras de esgoto por SPE-LC/QTOF. Benzoilecgonina foi usada para estimar o consumo de cocaína usando um fator de correção adaptado que considera as taxas de excreção de BE para diferentes vias de administração, bem como a fração de usuários de crack no Brasil. Análises diárias indicam um maior uso de cocaína nos finais de semana, com picos no domingo nas áreas sul (7385±121 mg dia-1 1000 hab-1) e norte (3566±171 mg dia-1 1000 hab-1) de Brasília. Um alto consumo de cocaínicos também foi observado no dia do Carnaval de 2018 na região norte (6229±219 mg dia-1 1000 hab-1). A razão COE/BE foi usada para avaliar o consumo simultâneo de cocaína e álcool. Razões maiores também foram observados durante o final de semana. No entanto, razões mais elevadas aos sábados do que aos domingos podem ser explicadas pela diminuição da excreção de BE durante o poliuso, o que pode levar a estimativas subestimadas de uso de cocaína aos sábados, bem como por diferentes padrões de uso de pó e crack, sendo que os últimos geralmente bebem quantidades menores de álcool. Os dados de epidemiologia comparados com as apreensões sugerem que a Polícia Civil de Brasília apreendeu apenas 3% de cocaína durante o estudo. As quantidades de adulterantes de cocaína, fenacetina e levamisol no esgoto foram relacionadas às amostras de crack e cocaína apreendidas pela Polícia Federal para estimar uma prevalência numérica de consumo entre uma ou outra via de administração. Estima-se que os usuários de crack no DF sejam responsáveis por cerca de 50% dos usuários de cocaínicos. Os dados gerados contribuem para tornar a ferramenta mais robusta e completa, além de continuar fornecendo informações úteis às iniciativas de enfrentamento ao uso e ao tráfico de drogas.The analysis of target molecules in a wastewater generated by a population has provided complementary information to the monitoring of drug use. This tool, commonly known in the literature as wastewater-based epidemiology has been used in the Brazilian Federal District in order to estimate the consumption of cocaine. Previously results showed that cocaine consumption doubles during weekends in Federal District suggesting the existence of different group users. One of the groups is composed of chemical dependents who constantly use drugs and the other one of intermittent users who may double cocaine consumption during the weekends. With the purpose of differentiating this kind of situation, an alternative would be monitoring the product of cocaine and alcohol metabolization called cocaethylene, which might indicate co-consumption of both substances. In this context, this work quantified cocaine, its metabolites benzoylecgonine and cocaethylene, plus its adulterants phenacetin and levamisole wastewater samples by SPE-LC/QTOF. Benzoylecgonine was used to estimate cocaine consumption using a country-tuned correction factor that considers BE excretion rates for different routes of administration as well as the fraction of crack users in Brazil. A day-to-day analysis indicates higher cocaine use in weekends with peaks on Sunday in both southern (7385±121 mg day-1 1000 inhab-1) and northern (3566±171 mg day-1 1000 inhab-1) areas of Brasilia. A significant high cocaine use was also observed in the Carnival Day of 2018 in the northern area (6229±219 mg day-1 1000 inhab-1). COE/BE ratios were used to assess cocaine and alcohol co- consumption since COE is produced during the polyuse of both substances. Higher ratios were also observed during the weekend. However, higher ratios on Saturdays rather than Sundays may be explained by the decrease of BE excretion during the polyuse, which may lead to an underestimation of the cocaine use estimates on Saturdays, as well as by different patterns of polyuse by powder and crack users, where the latter usually drink lower amounts of alcohol. The data compared to seized drugs suggests that the Brazilian Civil Police seized only 3% of cocaine during the study. The amounts of cocaine adulterants, phenacetin, and levamisole in the sewage were related to the crack and cocaine samples seized by the Federal Police to estimate a numerical prevalence of consumption between one or another route of administration. It has been estimated that crack cocaine users in the DF account for about 50% of cocaine users. The findings contribute to making the tool more robust and complete, as well as providing useful information for initiatives to combat drug use and trafficking

Assessing Cocaine Use Patterns in the Brazilian Capital by Wastewater-Based Epidemiology

This is the postprint (accepted manuscript) version of the article published by Taylor & Francis in the International Journal of Environmental Analytical Chemistry on December, 10th 2018. Available on-line: http://www.tandfonline.com//10.1080/03067319.2018.1554743The levels of cocaine (COC) and its metabolites benzoylecgonine (BE) and cocaethylene (COE) in wastewater samples from Brasilia, Brazil, were determined using liquid chromatography coupled to hybrid quadrupole-time-of-flight mass spectrometry. The former metabolite was used to estimate cocaine consumption using a country-tuned correction factor that considers BE excretion rates for different routes of administration as well as the fraction of crack users in Brazil. A day-to-day analysis during a week indicates higher cocaine use in weekends with peaks on Sunday in both southern (7385 ± 121 mg day−1 1000 inhab−1) and northern (3566 ± 171 mg day−1 1000 inhab−1) areas of Brasilia. A significant high cocaine use was also observed in the Carnival Day of 2018 (6229 ± 219 mg day−1 1000 inhab−1). COE/BE ratios were used to assess COC and alcohol co-consumption since COE is produced during the co-consumption of both substances. Higher ratios were also observed during the weekend. However, higher ratios on Saturdays rather than Sundays may be explained by the decrease of BE excretion during the co-consumption with alcohol, which may lead to an underestimation of the cocaine use estimates on Saturdays, as well as by different patterns of co-consumption by powder and crack users, where the latter usually drink lower amounts of alcohol. Our data suggest that only 3% of cocaine was seized by the local Technical Police during the studyThis work was supported by the Federal District Research Foundation under Grant [193.000.916/ 2015]; the Spanish Agencia Estatal de Investigación under Grant [CTM2017-84763-C3-2-R]; the Galician Council of Culture, Education and Universities under Grants [ED431C2017/36]; IGM postdoctoral contract, Plan Galego I2C-Modalidade B, ED481D 2017/003; and FEDER/ERDFS

Profiling cocaine residues and pyrolytic products in wastewater by mixed‐mode liquid chromatography–tandem mass spectrometry

This is the peer reviewed version of the following article: González‐Mariño, I, Estévez‐Danta, A, Rodil, R, et al. Profiling cocaine residues and pyrolytic products in wastewater by mixed‐mode liquid chromatography–tandem mass spectrometry. Drug Test Anal. 2019; 11: 1018– 1027, which has been published in final form at https://doi.org/10.1002/dta.2590. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived VersionsThis work provides a new analytical method for the determination of cocaine, its metabolites benzoylecgonine and cocaethylene, the pyrolytic products anhydroecgonine and anhydroecgonine methyl ester, and the pharmaceutical levamisole in wastewater. Samples were solid‐phase extracted and extracts analyzed by liquid chromatography–tandem mass spectrometry using, for the first time in the illicit drug field, a stationary phase that combines reversed‐phase and weak cation‐exchange functionalities. The overall method performance was satisfactory, with limits of detection below 1 ng/L, relative standard deviations below 21%, and percentages of recovery between 93% and 121%. Analysis of 24‐hour composite raw wastewater samples collected in Santiago de Compostela (Spain) and Brasilia (Brazil) highlighted benzoylecgonine as the compound showing the highest population‐normalized mass loads (300–1000 mg/day/1000 inhabitants). In Brasilia, cocaine and levamisole loads underwent an upsurge on Sunday, indicating a high consumption, and likely a direct disposal, of cocaine powder on this day. Conversely, the pyrolytic product resulting from the smoke of crack, anhydroecgonine methyl ester, and its metabolite anhydroecgonine were relatively stable over the four days, agreeing with a non‐recreational‐associated use of crackThis work was financially supported by Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED431C2017/36 and IGM postdoctoral contract, Plan Galego I2C‐Modalidade B, ED481D 2017/003), Ministerio de Economía y Competitividad (project no. CTM2017‐84763‐C3‐2‐R), the Federal District Research Foundation (project no. 193.000.916/2015), and FEDER/ERDFS

Mass Spectrometry-Based Zebrafish Toxicometabolomics: A Review of Analytical and Data Quality Challenges

Metabolomics has achieved great progress over the last 20 years, and it is currently considered a mature research field. As a result, the number of applications in toxicology, biomarker, and drug discovery has also increased. Toxicometabolomics has emerged as a powerful strategy to provide complementary information to study molecular-level toxic effects, which can be combined with a wide range of toxicological assessments and models. The zebrafish model has gained importance in recent decades as a bridging tool between in vitro assays and mammalian in vivo studies in the field of toxicology. Furthermore, as this vertebrate model is a low-cost system and features highly conserved metabolic pathways found in humans and mammalian models, it is a promising tool for toxicometabolomics. This short review aims to introduce zebrafish researchers interested in understanding the effects of chemical exposure using metabolomics to the challenges and possibilities of the field, with a special focus on toxicometabolomics-based mass spectrometry. The overall goal is to provide insights into analytical strategies to generate and identify high-quality metabolomic experiments focusing on quality management systems (QMS) and the importance of data reporting and sharing

Combined western diet and bisphenol A exposure induces an oxidative stress-based paraoxonase 1 response in larval zebrafish

<p>Paraoxonase 1 (PON1) is an antioxidant enzyme linked to metabolic disorders by genome-wide association studies in humans. Exposure to metabolic disrupting chemicals (MDCs) such as bisphenol A (BPA), together with genetic and dietary factors, can increase the risk of metabolic disorders. The objective of this study was to investigate how PON1 responds to the metabolic changes and oxidative stress caused by a western diet, and whether exposure to BPA alters the metabolic and PON1 responses. Zebrafish larvae at 14 days post fertilization were fed a custom-made western diet with and without aquatic exposure to two concentrations of BPA for 5 days. A combination of western diet and 150 μg/L BPA exposure resulted in a stepwise increase in weight, length and oxidative stress, suggesting that BPA amplifies the western diet-induced metabolic shift. PON1 arylesterase activity was increased in all western diet and BPA exposure groups and PON1 lactonase activity was increased when western diet was combined with exposure to 1800 μg/L BPA. Both PON1 activities were positively correlated to oxidative stress. Based on our observations we hypothesize that a western diet caused a shift towards fatty acid-based metabolism, which was increased by BPA exposure. This shift resulted in increased oxidative stress, which in turn was associated with a PON1 activity increase as an antioxidant response. This is the first exploration of PON1 responses to metabolic challenges in zebrafish, and the first study of PON1 in the context of MDC exposure in vertebrates.</p&gt