RESUMO
Brazilian green propolis is used in folk medicine because of its various biological properties. The hydroalcoholic extract of Brazilian green propolis is characteristic for possessing several pharmacological properties. Phytochemical investigations have attributed some of these properties to the presence of compounds, which were chosen as analytical markers. This paper reports the development and analytical validation using UPLC-MS/MS in MRM mode. Veratraldehyde was used as an internal standard in qualitative and quantitative analyses of the extracts. Relative standard deviation values obtained for intra-day and inter-day precision were lower than 4%. Of the five parameters for robustness, wavelength detection and flow rate were the critical ones. Limits of detection and quantification ranged from 0.300 to 39.500 ng.mL-1 and from 1.400 to 85.00 ng.mL-1, respectively. The recoveries were between 94.00 and 119.00%, with relative standard deviation values around 5.0%. The developed method is precise, sensitive, and reliable for analysing Brazilian green propolis.
RESUMO
Exposure to mycotoxins through food is a major health concern, especially for youngsters. This study performed a preliminary investigation on children's exposure to dietary mycotoxins in Ribeirão Preto, Brazil. Sampling procedures were conducted between August and December 2022, to collect foods (N = 213) available for consumption in the households of children (N = 67), including preschoolers (aged 3-6 years, n = 21), schoolers (aged 7-10 years, n = 15), and adolescents (aged 11-17 years, n = 31) cared in the Vila Lobato Community Social Medical Center of Ribeirão Preto. Ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) was used to determine concentrations of the mycotoxins in foods. Mycotoxins measured in all foods comprised aflatoxins (AFs), fumonisins (FBs), zearalenone (ZEN), T-2 toxin, deoxynivalenol (DON) and ochratoxin A (OTA). Higher incidence and levels were found for FBs, ZEN, and DON in several commonly consumed foods. Furthermore, 32.86 % foods had two to four quantifiable mycotoxins in various combinations. The mean estimated daily intake (EDI) values were lower than the tolerable daily intake (TDI) for AFs, FBs, and ZEN, but higher than the TDI (1.0 µg/kg bw/day) for DON, hence indicating a health risk for all children age groups. Preschoolers and adolescents were exposed to DON through wheat products (EDIs: 2.696 ± 7.372 and 1.484 ± 2.395 µg/kg body weight (bw)/day, respectively), while schoolers were exposed through wheat products (EDI: 1.595 ± 1.748 µg/kg bw/day) and rice (EDI: 1.391 ± 1.876 µg/kg bw/day). The results indicate that wheat-based foods and rice may be risky to children, implying the need for stringent measures to avoid DON contamination in these products.
Assuntos
Aflatoxinas , Micotoxinas , Zearalenona , Criança , Adolescente , Humanos , Micotoxinas/análise , Projetos Piloto , Cromatografia Líquida/métodos , Brasil , Contaminação de Alimentos/análise , Espectrometria de Massas em Tandem/métodos , Zearalenona/análise , Aflatoxinas/análise , TriticumRESUMO
Despite potential health implications, data on the presence of Glyphosate (GLY) and other non-GLY herbicides in human matrices remain scarce. This study aimed to develop a simple and cost-effective methodology for detecting and quantifying GLY, its primary biodegradation product; aminomethylphosphonic acid (AMPA); and glufosinate (GLU) in plasma and urine of environmentally and occupationally exposed populations from the province of Córdoba (Argentina). Different alternatives of pre-treatment, derivatization with FMOC-Cl, solid phase extraction, and final sample conditioning steps were evaluated to improve the quantification of the herbicides by a high-performance liquid chromatography system coupled to a triple-quadrupole mass spectrometer. Recoveries ranged from 39 to 84% in both matrices, while limits of quantification were 3, 1, and 0.3 ng/mL and 3.6, 5.1, and 0.3 ng/mL for AMPA, GLY, and GLU in plasma and urine, respectively. In plasma samples, GLY was the most frequently detected analyte (32%), followed by GLU (10%). In urine samples, GLU was the most frequently detected herbicide (13%), followed by GLY (6%). No differences between group or matrix correlations were found. This study is the first report of GLU in human biological matrices and should be used to establish baseline values for future surveillance systems.
RESUMO
Tuberculosis (TB) is a high-burden infectious disease with high prevalence and mortality rates. The first-line anti-TB drugs include isoniazid (INH), rifampicin (RMP), pyrazinamide (PZA), and ethambutol (EMB). At present, the standard method of blood sampling for therapeutic drug monitoring (TDM) analysis is venipuncture. Dried blood spots (DBS) are a minimally invasive method for collecting small quantities of whole blood from fingertips. The aim of the current study was to develop an ultrahigh-performance liquid chromatography technique coupled to tandem mass spectrometry (UPLC-MS/MS) for simultaneous quantification of the first-line anti-TB drugs in human plasma and DBS as a sampling alternative. The separation and detection conditions were optimized to quantify INH, RMP, PZA, and EMB in both matrices in an ACQUITY UPLC H Class system coupled to a XEVO TQD detector. Chromatographic separation was performed through an Acquity HSS T3 column (2.1 × 100 mm, 1.8 µm) with 0.1% formic acid in water and acetonitrile as the mobile phase. The total run time was 7 min for both methods, with retention time in plasma of 0.85, 1.22, 3.16, and 4.04 min and 0.74, 0.87, 0.97, and 4.16 min for EMB, INH, PZA, and RMP in DBS, respectively. The bioanalytical methods developed were proved selective, linear, precise, and accurate (inter- and intra-assay); the matrix effect was demonstrated to be within the established limits. Short- and long-term stability, freeze-thaw cycles for plasma, and short-term stability for DBS were established. A total of 15 patients with 46 ± 17 (mean ± SD) years old were included, and anti-TB drug concentrations were quantified on plasma and DBS as proof of concept. Based on RMP and INH plasma concentrations (Cp), and Bayesian estimation of individual pharmacokinetic parameters, a dose adjustment was necessary for 93% of patients. The slopes of the correlation lines between plasma and DBS concentrations of RMP, EMB, INH, and PZA were 0.5321, 0.8125, 0.5680, and 0.6791, respectively. Finally, significant correlations (p < 0.05) were observed between DBS and plasma concentrations for RMP (r2 = 0.6961), EMB (r2 = 0.4369), INH (r2 = 0.8675) and PZA (r2 = 0.7363). A simple, fast, and reliable UPLC-MS/MS method was developed to quantify first-line anti-TB drugs in plasma and DBS, which provides an easy sampling and storage to be applied as a new strategy for TDM in patients with TB.
Assuntos
Antituberculosos , Tuberculose , Humanos , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida/métodos , Cromatografia Líquida de Alta Pressão/métodos , Teorema de Bayes , Tuberculose/tratamento farmacológico , Isoniazida , Rifampina , Etambutol , Padrões de ReferênciaRESUMO
Background: Workplace drug testing primarily relies on urine analysis, targeting multiple compounds with varying physicochemical characteristics. Biocompatible solid-phase microextraction (BioSPME) is a miniaturized solid-phase extraction technique that enables the simultaneous extraction and preconcentration of analytes directly from the biological matrix. Methods: The BioSPME procedure consisted of the sequential extraction of 50-µl urine samples using LC Tips C18 in basic and acidic pH, followed by desorption with methanol and n-hexane, respectively. The extracts were analyzed by ultra-performance LC-MS/MS. Results: Intra-day precision was 1.2-8.6% and inter-day precision was 1.8-14.2%. Accuracy was 96.8-107.4%. The extraction yields were 62.8-109.4%. The matrix effects were -3.98% to 1%. Conclusion: BioSPME shows promise as an alternative method for preparing urine samples prior to drug measurement by ultra-performance LC-MS/MS.
Assuntos
Cocaína , Dronabinol , Cromatografia Líquida/métodos , Cocaína/análise , Analgésicos Opioides , Espectrometria de Massas em Tandem/métodos , Extração em Fase Sólida , Anfetaminas/análise , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Este estudio se realizó con el objetivo de desarrollar y validar un método para la determinación de 30 medicamentos veterinarios en muestras de trucha y langostino. El método utiliza extracción en fase sólida dispersiva (dSPE) con C18 y detección por cromatografía líquida acoplada a espectrometría de masas. Se determinó linealidad, veracidad (porcentaje de recuperación), repetitividad y reproducibilidad intralaboratorio (porcentaje de desviación estándar relativa (% RSD)), límites de detección (LoD), límites de cuantificación (LoQ), selectividad e incertidumbre. La recuperación varió de 70 a 120% y la repetibilidad y la reproducibilidad fueron menores de 20% de la desviación estándar relativa. La selectividad fue adecuada, sin picos interferentes. Las relaciones iónicas cumplieron con los criterios de confirmación. Los coeficientes de determinación (R2) fueron mayores de 0,99, con excepción de la sulfaquinoxalina en langostino (R2 = 0,97). Los LoD y los LoQ variaron entre 0,6 µg/kg y 12,8 µg /kg y los valores de incertidumbre entre 6 µg/kg y 49 µg/ kg. Se analizaron adicionalmente 6 muestras de diferentes mercados de Lima y se detectaron trazas de algunos medicamentos incluidos en el ensayo. El método es adecuado para el análisis de residuos de medicamentos veterinarios y se recomienda su aplicación en los programas nacionales de monitoreo de la inocuidad de truchas y langostinos provenientes de acuicultura.
The study was aimed at developing and validate an analysis method to determine residues of 30 veterinary drugs in rainbow trout and shrimp specimens. The method involves extraction in dispersive solid phase with C18 and the subsequent detection through liquid chromatography coupled to mass spectrometry. Validation was done through determination of linearity, trueness (% of recovery), repeatability and intralaboratory reproducibility, limits of detection (LoD), limits of quantification (LoQ) selectivity and uncertainty. Recovery ranged from 70 to 120% and repeatability and intralaboratory reproducibility were lower than 20%. Selectivity was adequate, without interference peaks. Likewise, the ionic relationships met the confirmation criteria. The linearity was adequate, with determination coefficients (R2) above 0.99, except for sulfaquinolaxin in shrimp specimens (R2 = 0,97). LoD and LoQ varied from 0,6 µg /kg to 12,8 µg / kg. Limits of uncertainty ranged from 6 µg /kg to 49 µg /kg. The method was used to analyze 6 samples from different markets in Lima (Peru), identifying traces of some drugs included in the study. Our results show that the method is adequate for the analysis of veterinary drug residues and allow us to recommend its application in national monitoring programs, to assess the safety of rainbow trout and shrimp specimens from aquaculture.
O estudo foi realizado com o objetivo de desenvolver e validar um método para a determinação de 30 medicamentos veterinários, em amostras de truta e camarão. O método utiliza extração dispersiva em fase sólida com C18 e detecção por cromatografia líquida acoplada à espectrometria de massas. Foram determinados a linearidade, a veracidade (recuperação percentual), a repetibilidade, a reprodutibilidade intra-laboratorial, os limites de detecção (LoD) e de quantificação (LoQ), a linearidade, a selectividade e a incerteza. A recuperação variou de 70 a 120%, a repetibilidade e reprodutibilidade estiveram abaixo do 20% do desvio padrão relativo. A selectividade fio adequada, sem picos de interferentes. As proporções de íons atenderam aos critérios de confirmação. Os coeficientes de determinação (R2) foram superiores a 0,99, com excepção da sulfanoxalina em camarão (R2 = 0,97). LoD e LoQ variavam entre 0,6 µg /kg e 12,8 µg /kg e valores de incerteza entre 6 µg /kg e 49 µg / kg. Seis amostras de mercados do Lima foram adicionalmente analisadas e foram detectados vestígios de alguns medicamentos incluídos no estudo. O método é adequado para o análise de resíduos de medicamentos veterinários e sua aplicação é recomendada em programas nacionais de controlo da segurança da truta e do camarão provenientes da aquicultura.
RESUMO
This work explores the degradation of xenobiotic compounds in aerobic and anaerobic batch reactors. Different inoculums were spiked with nine emerging contaminants at nominal concentrations ranging between 1 to 2 mg/L (ibuprofen, diclofenac, naproxen, acesulfame, sucralose, aspartame, cyclamate, linear alkylbenzene sulfonates, and secondary alkyl sulfonates). Ethanol was used as co-substrate in the anaerobic reactors. We found that the kinetic decay was faster in the aerobic reactors inoculated with a Spanish (Spn) inoculum compared to a Brazilian (Brz) inoculum, resulting in rection rates for LAS and SAS of 2.67 ± 3.6 h-1 and 5.09 ± 6 h-1 for the Brz reactors, and 1.3 ± 0.1 h-1 and 1.5 ± 0.2 h-1 for the Spn reactors, respectively. There was no evidence of LAS and SAS degradation under anaerobic conditions within 72 days; nonetheless, under aerobic conditions, these surfactants were removed by both the Brz and Spn inoculums (up to 86.2 ± 9.4% and 74.3 ± 0.7%, respectively) within 10 days. The artificial sweeteners were not removed under aerobic conditions, whereas we could observe a steady decrease in the anaerobic reactors containing the Spn inoculum. Ethanol aided in the degradation of surfactants in anaerobic environments. Proteiniphilum, Paraclostridium, Arcobacter, Proteiniclasticum, Acinetobacter, Roseomonas, Aquamicrobium, Moheibacter, Leucobacter, Synergistes, Cyanobacteria, Serratia, and Desulfobulbus were the main microorganisms identified in this study.
Assuntos
Esgotos , Tensoativos , Esgotos/química , Anaerobiose , Biodegradação Ambiental , Tensoativos/metabolismo , Etanol , Reatores BiológicosRESUMO
Taperebá (Spondias mombin L.) is a native species of the Brazilian Cerrado that has shown important characteristics such as a significant phenolic compound content and biological activities. The present study aimed to characterize the phenolic compound profile and antioxidant activity in taperebá peel extract, as well as microencapsulating the extract with chitosan and evaluating the stability of the microparticles. The evaluation of the profile of phenolic compounds was carried out by UPLC-MS/MS. The in vitro antioxidant activity was evaluated by DPPH and ABTS methods. The microparticles were obtained by spray drying and were submitted to a stability study under different temperatures. In general, the results showed a significant content of polyphenols and antioxidant activity. The results of UPLC-MS/MS demonstrated a significant content of polyphenols in taperebá peel, highlighting the high content of ellagic acid and quercetin compounds. There was significant retention of phenolic compounds when microencapsulated, demonstrating high retention at all evaluated temperatures. This study is the first to microencapsulate the extract of taperebá peel, in addition to identifying and quantifying some compounds in this fruit.
Assuntos
Anacardiaceae , Quitosana , Anacardiaceae/química , Antioxidantes/química , Brasil , Quitosana/análise , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Frutas/química , Fenóis/química , Extratos Vegetais/química , Polifenóis/análise , Espectrometria de Massas em TandemRESUMO
An analytical method of ultra-high performance liquid chromatography coupled to tandem mass spectrometry detection was developed and validated for the simultaneous quantification in plasma of four selective serotonin reuptake inhibitor antidepressants: sertraline, escitalopram, paroxetine, fluoxetine, and its metabolite norfluoxetine. A simple protein precipitation was performed with acetonitrile containing 100 ng/mL of indomethacin, which was used as internal standard. Chromatographic separation was carried out on an Acquity BEH C18 column with isocratic elution of the mobile phase consisting of 5 mmol/L ammonium acetate with 0.1% formic acid (A) and acetonitrile (B) at a 60:40 proportion, respectively. The flow rate was 0.4 mL/min with a run time of 5 min. A positive electrospray ionization source was used for detection. The method was linear in a range of 5-800 ng/mL, with determination coefficients greater than 0.991. The accuracy ranged from 91% to 112% for intra-assay and from 89% to 112% for inter-assay. The variation coefficients ranged from 3.1% to 14.88% for intra-assay and from 3.60% to 14.74% for inter-assay precision. The method was successfully applied for the analysis of 73 samples from patients under treatment with these antidepressants; 36.9% of the samples had concentrations outside therapeutic ranges. This method can be applied for routine analysis in clinical practice, simplifying sample processing, reducing analysis time and consequently the costs associated with it.
RESUMO
O número de pessoas utilizando substâncias ilícitas de forma recreativa aumenta a cada ano, chamando a atenção de estudiosos de diversas áreas do conhecimento. Com isso, a demanda de exames toxicológicos exigida para trabalhadores, vítimas de crimes e esportistas também tem crescido. A amostra biológica mais utilizada para análises toxicológicas continua sendo a urina, visto que sua obtenção é menos invasiva, possibilita coletar grande volume de amostra e pode-se detectar substâncias até dias após ter ocorrido a exposição ou consumo. Entretanto, estas amostras necessitam de um grande volume físico para serem armazenadas e transportadas aos laboratórios, devendo ser mantidas em temperatura baixa e controlada para conservação. Outro ponto a se considerar é a quantidade de amostra insuficientemente coletada, ou extravasamento do conteúdo, contaminando outras amostras e muitas vezes, inviabilizando a análise. Uma alternativa recente para tais problemas é utilizar a técnica chamada de dried urine spots (DUS), onde poucos microlitros de urina são colocados em um papel absorvente e secos sob temperatura ambiente, preservando de agentes degradantes os componentes presentes na urina. Assim, o objetivo deste trabalho é avaliar a estabilidade das substâncias do presente estudo em alta temperatura, temperatura ambiente e em temperaturas de 4°C e -20°C. Para este fim, foi necessário desenvolver, validar e aplicar métodos de extração e determinação de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol carboxílico (THCCOOH) em amostras dried urine spot, utilizando cromatografia líquida acoplada à espectrometria de massas. Os picos foram identificados por UPLC-ESI-MS/MS, com tempo total de 5 mins utilizando fase A- água, formiato de amônio e 0,1% ácido fórmico, e B- metanol: acetonitrila (6:4) + 0,1% de ácido fórmico. A extração foi feita utilizando acetonitrila: metanol: acetona (1:1:1) +ácido fórmico 0,1%. Não foi possível iniciar a validação de THCCOOH, visto uma possível complexação do analito com o papel. Para as outras substâncias, o método cromatográfico desenvolvido se mostrou eficiente e seletivo, com LOD e LOQ de 10 ng/mL para todos os analitos, sendo linear até 1000 ng/mL, atendeu as especificações de precisão e exatidão e carryover. As amostras permaneceram estáveis ao longo de 32 dias nas temperaturas estudadas, demonstrando a segurança em se utilizar a técnica de DUS para armazenamento e transporte de amostras biológicas dentro da faixa de temperatura do estudo até 32 dias
The number of people using illegal substances in a recreational way increases each year, drawing the attention of scholars from different areas of knowledge. As a result, the demand for workplaces drug tests, toxicological tests for victims of crimes and dopping has also grown. The biological sample most used for toxicological tests remains urine, since obtaining it is less invasive, it is possible to collect a large volume of sample and it is possible to detect substances up to days after exposure or consumption has occurred. However, these samples require a large physical volume to be stored and transported to the laboratories, and must be kept at a low temperature for conservation. Another point to consider is the amount of sample insufficiently collected, or leakage of the content, causing contamination of other samples and often making the analysis unfeasible. A recent alternative to such problems is to use "dried urine spots" (DUS), where few microliters of urine are placed on absorbent paper and dried at room temperature, preserving the components present in the urine from degrading agents. Thus, the objective of this work is to evaluate the stability of the substances in this study at high temperature, room temperature and at temperatures of 4°C and -20°C. For this purpose, it was necessary to develop, validate and apply methods of extraction and determination of amphetamines and biotransformation products of cocaine and carboxylic tetrahydrocannabinol (THCCOOH) in dried urine spot samples, using liquid chromatography coupled to mass spectrometry (LC-MS). The peaks were identified liquid chromatography coupled to a mass spectrometer (UPLC-ESI-MS/MS), with a total time of 5 mins using phase A- water, ammonium formate and 0.1% formic acid, and B- methanol: acetonitrile (6:4) + 0.1% formic acid. Extraction was done using acetonitrile: methanol: acetone (1:1:1) + 0.1% formic acid. It was not possible to perform the validation of THCCOOH, given a possible complexation of the analyte with the paper. To the others substances, the chromatographic method developed proved to be efficient and selective, with LOD and LOQ of 10 ng/mL for all analytes, being linear up to 1000 ng/mL, meeting the specifications of precision and accuracy and carryover. The samples remained stable for 32 days at the temperatures studied, demonstrating the safety of using the DUS technique for storage and transport of biological samples until 32 days on temperature range studied
Assuntos
Dronabinol/efeitos adversos , Biotransformação , Cocaína/efeitos adversos , Anfetaminas/efeitos adversos , Espectrometria de Massas/métodos , Urina , Preparações Farmacêuticas/administração & dosagem , Cromatografia Líquida/métodos , Categorias de Trabalhadores/classificaçãoRESUMO
O número de pessoas utilizando substâncias ilícitas de forma recreativa aumenta a cada ano, chamando a atenção de estudiosos de diversas áreas do conhecimento. Com isso, a demanda de exames toxicológicos exigida para trabalhadores, vítimas de crimes e esportistas também tem crescido. A amostra biológica mais utilizada para análises toxicológicas continua sendo a urina, visto que sua obtenção é menos invasiva, possibilita coletar grande volume de amostra e pode-se detectar substâncias até dias após ter ocorrido a exposição ou consumo. Entretanto, estas amostras necessitam de um grande volume físico para serem armazenadas e transportadas aos laboratórios, devendo ser mantidas em temperatura baixa e controlada para conservação. Outro ponto a se considerar é a quantidade de amostra insuficientemente coletada, ou extravasamento do conteúdo, contaminando outras amostras e muitas vezes, inviabilizando a análise. Uma alternativa recente para tais problemas é utilizar a técnica chamada de dried urine spots (DUS), onde poucos microlitros de urina são colocados em um papel absorvente e secos sob temperatura ambiente, preservando de agentes degradantes os componentes presentes na urina. Assim, o objetivo deste trabalho é avaliar a estabilidade das substâncias do presente estudo em alta temperatura, temperatura ambiente e em temperaturas de 4°C e -20°C. Para este fim, foi necessário desenvolver, validar e aplicar métodos de extração e determinação de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol carboxílico (THCCOOH) em amostras dried urine spot, utilizando cromatografia líquida acoplada à espectrometria de massas. Os picos foram identificados por UPLC-ESI-MS/MS, com tempo total de 5 mins utilizando fase A- água, formiato de amônio e 0,1% ácido fórmico, e B- metanol: acetonitrila (6:4) + 0,1% de ácido fórmico. A extração foi feita utilizando acetonitrila: metanol: acetona (1:1:1) +ácido fórmico 0,1%. Não foi possível iniciar a validação de THCCOOH, visto uma possível complexação do analito com o papel. Para as outras substâncias, o método cromatográfico desenvolvido se mostrou eficiente e seletivo, com LOD e LOQ de 10 ng/mL para todos os analitos, sendo linear até 1000 ng/mL, atendeu as especificações de precisão e exatidão e carryover. As amostras permaneceram estáveis ao longo de 32 dias nas temperaturas estudadas, demonstrando a segurança em se utilizar a técnica de DUS para armazenamento e transporte de amostras biológicas dentro da faixa de temperatura do estudo até 32 dias
The number of people using illegal substances in a recreational way increases each year, drawing the attention of scholars from different areas of knowledge. As a result, the demand for workplaces drug tests, toxicological tests for victims of crimes and dopping has also grown. The biological sample most used for toxicological tests remains urine, since obtaining it is less invasive, it is possible to collect a large volume of sample and it is possible to detect substances up to days after exposure or consumption has occurred. However, these samples require a large physical volume to be stored and transported to the laboratories, and must be kept at a low temperature for conservation. Another point to consider is the amount of sample insufficiently collected, or leakage of the content, causing contamination of other samples and often making the analysis unfeasible. A recent alternative to such problems is to use "dried urine spots" (DUS), where few microliters of urine are placed on absorbent paper and dried at room temperature, preserving the components present in the urine from degrading agents. Thus, the objective of this work is to evaluate the stability of the substances in this study at high temperature, room temperature and at temperatures of 4°C and -20°C. For this purpose, it was necessary to develop, validate and apply methods of extraction and determination of amphetamines and biotransformation products of cocaine and carboxylic tetrahydrocannabinol (THCCOOH) in dried urine spot samples, using liquid chromatography coupled to mass spectrometry (LC-MS). The peaks were identified liquid chromatography coupled to a mass spectrometer (UPLC-ESI-MS/MS), with a total time of 5 mins using phase A- water, ammonium formate and 0.1% formic acid, and B- methanol: acetonitrile (6:4) + 0.1% formic acid. Extraction was done using acetonitrile: methanol: acetone (1:1:1) + 0.1% formic acid. It was not possible to perform the validation of THCCOOH, given a possible complexation of the analyte with the paper. To the others substances, the chromatographic method developed proved to be efficient and selective, with LOD and LOQ of 10 ng/mL for all analytes, being linear up to 1000 ng/mL, meeting the specifications of precision and accuracy and carryover. The samples remained stable for 32 days at the temperatures studied, demonstrating the safety of using the DUS technique for storage and transport of biological samples until 32 days on temperature range studied
Assuntos
Dronabinol/efeitos adversos , Biotransformação , Cocaína/agonistas , Anfetaminas/análise , Espectrometria de Massas/métodos , Urina/fisiologia , Cromatografia Líquida/métodosRESUMO
Antimicrobials premixes are the presentation of choice to administer drugs simultaneously to groups of animals in intensive husbandry systems that require treatment for pathologies of bacterial origin. Among the premixes available for use in poultry, florfenicol and oxytetracycline are commonly administered via food or water. However, their actual concentration in premixes must meet on-label statements to ensure plasma concentrations reach effective therapeutic levels. Hence, this work was designed for the purpose of verifying whether the concentration of antimicrobial present in five premixes matched their on-label statement. Three oxytetracycline premixes, and two of florfenicol, were analysed using a Xevo TQ-S micro UPLC-MS/MS, and an ABSciex API4000 HPLC-MS/MS, respectively. Analytical methodologies were implemented and validated, showing an R2 ≥ 0.99 for the calibration curves. Oxytetracycline was detected in these premixes at concentrations exceeding on-label statements by 13.28%, 21.54%, and 29.68%, whereas florfenicol concentrations detected in premixes were 13.06% and 14.75% lower than expected. Consequently, this work shows that the concentration of active ingredients that are present in commercial formulations effectively differ from those stated on premix labels, and it also highlights how unpredictable their range of variability might be. This must be addressed through solid and updated laws that guarantee an effective pharmaceutical product.
RESUMO
The goal of this survey was to evaluate the presence and concentration as well as the co-occurrence of legislated and non-legislated mycotoxins in wheat flour samples from Brazil. A total of 200 wheat flour samples were analysed by a validated multi-mycotoxins method. DON was the mycotoxin with the highest occurrence, being present in 100% of the analysed samples and showing contamination in both years and regions (53-2905 µgâ¯kg-1). ZEN was detected in 51% (Assuntos
Micotoxinas
, Tricotecenos
, Brasil
, Farinha/análise
, Contaminação de Alimentos/análise
, Micotoxinas/análise
, Tricotecenos/análise
, Triticum
RESUMO
Misoprostol is a prostaglandin E1 synthetic analogous used for elective interruptions of early pregnancy, treatment of incomplete abortion, postpartum hemorrhage and induction of full-term labor. Its a lipophilic drug, passing by extensive and rapid pre-systemic metabolism into the active metabolite, misoprostol acid (MA). The objective of this study was to develop and validate a highly sensitive method for MA determination in plasma using UPLC-MSMS, with application in a study of maternal-fetal pharmacokinetics in healthy parturients women (n = 10) after administration of 25 µg misoprostol vaginally. The method presented linearity of 2-10 pg/mL and acceptable precision, accuracy, plasma and solution stability. The parturients women presented median (interquartile range) values of AUC0-6 of 68.0 (40.8-84.7) pg.h/mL, Cmax of 21.9 (11.9-30.1) pg/mL and Tmax of 2.25 (0.69-5.00) h. The placental transfer of MA was assessed from the umbilical vein/maternal blood ratios of 1.40 (0.91-2.13) and intervillous space/maternal blood ratios of 0.49 (0.15-3.41). In conclusion, this method presented high sensitivity, being able to quantify MA in plasma samples following a low 25 µg misoprostol administered vaginally aimed to induce labor in parturients women. Additionally, this is the first description of the placental transfer of MA after a vaginal administration of misoprostol.
Assuntos
Misoprostol , Administração Intravaginal , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Feminino , Humanos , Trabalho de Parto Induzido , Misoprostol/análogos & derivados , Placenta , Gravidez , Espectrometria de Massas em TandemRESUMO
The determination of psychotropic drugs and metabolites in blood is relevant in the context of both therapeutic drug monitoring and clinical and forensic toxicology. LC-MS/MS is the preferred method for these assays. However, LC-MS/MS is particularly susceptible to matrix ionization effects and appropriate sample preparation is required to minimize these effects. In this study, a simple, single-step, mini-QuEchERS extraction procedure, coupled to UPLC-MS/MS, was developed and validated for the determination of 15 toxicologically relevant compounds in whole blood, including psychoactive drugs and some metabolites. The assay was linear in the range of 25-1,000 ng ml-1 , fulfilling criteria for accuracy and precision. Extraction yields (71.9-87.7%) and matrix effects (-3.3 to +4.4%, with the exception of codeine, which had matrix effects of -35.36 to -28.14%) were acceptable for the majority of the evaluated compounds, using a single internal standard. The assay was applied to 238 clinical specimens from patients admitted to an emergency service, with 22 samples presenting quantifiable concentrations of 11 different compounds. The developed assay is a simple and efficient strategy for determination of target psychotropic drugs and metabolites in forensic and clinical toxicology.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Psicotrópicos , Espectrometria de Massas em Tandem/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Psicotrópicos/sangue , Psicotrópicos/isolamento & purificação , Psicotrópicos/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Adulto JovemRESUMO
Methotrexate is the gold standard treatment in rheumatoid arthritis. Once absorbed, it is internalized in cells, where glutamate residues are added to produce polyglutamated forms, which are responsible for the effect of methotrexate. The aim of the current study is to determine the relationship between methotrexate triglutamate concentrations and the clinical evolution in rheumatoid arthritis patients, as well as to characterize the variability in both features to propose strategies for low-dose methotrexate optimization. The quantification of methotrexate triglutamate concentration in red blood cells was performed through ultra-performance liquid chromatography coupled with mass spectrometry. Polymorphisms of genes involved in the formation of polyglutamates were determined by real-time polymerase chain reaction. A multivariate regression was performed to determine the covariates involved in the variability of methotrexate triglutamate concentrations and a population pharmacokinetics model was developed through nonlinear mixed-effects modeling. Disease activity score changed according to methotrexate triglutamate concentrations; patients with good response to treatment had higher concentrations than moderate or nonresponding patients. The methotrexate triglutamate concentrations were related to time under treatment, dose, red blood cells, and body mass index. A 1-compartment open model was selected to estimate the pharmacokinetic parameters; the typical total clearance (L/day) was determined as 1.45 * (body mass index/28 kg/m2 ) * (red blood cells/4.6 × 106 cells/µL) and the volume of distribution was 52.4 L, with an absorption rate of 0.0346/day and a fraction metabolized of 1.03%. Through the application of the model, the initial dose of methotrexate is proposed on the basis of stochastic simulations and considering methotrexate triglutamate concentrations found in responders patients.
Assuntos
Antirreumáticos/farmacocinética , Artrite Reumatoide/tratamento farmacológico , Metotrexato/análogos & derivados , Ácido Poliglutâmico/análogos & derivados , Fatores Etários , Antirreumáticos/sangue , Antirreumáticos/uso terapêutico , Índice de Massa Corporal , Peso Corporal , Relação Dose-Resposta a Droga , Eritrócitos , Genótipo , Humanos , Estudos Longitudinais , Taxa de Depuração Metabólica , Metotrexato/sangue , Metotrexato/farmacocinética , Metotrexato/uso terapêutico , México , Modelos Biológicos , Ácido Poliglutâmico/sangue , Ácido Poliglutâmico/farmacocinética , Ácido Poliglutâmico/uso terapêutico , Polimorfismo de Nucleotídeo Único , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Zilpaterol and Clenbuterol are ß-adrenergic agonists that have been widely used to feed cattle. Although the use of Zilpaterol has been approved, Clenbuterol is still used illegally at unknown doses. However, the research of both substances has been based mainly on the evaluation of residues. To our knowledge, this is the first time that a cellular model using Hep G2 cells treated with Zilpaterol and Clenbuterol is presented as an alternative approach to quantify both drugs at the cellular level. Thus, a complete analytical methodology has been developed for the accurate quantitation of these ß-adrenergic agonists in both cellular compartments. We propose the use of ultra-performance liquid chromatography with photodiode array detector (UPLC-PDA) for extracellular determinations while UPLC coupled to a tandem mass spectrometer (UPLC-MS/MS) for intracellular analysis. The methods were fully validated in terms of selectivity, linearity, accuracy, and precision, limits of detection and quantitation (LOD and LOQ, respectively), stability, carryover, and matrix effect. The method for intracellular content was linear ranging from 0.25 to 8 ng/mL while for extracellular content, the concentration of Zilpaterol and Clenbuterol ranged from 0.125 to 4 µg/mL, with correlation coefficients of R > 0.98 and >0.99, respectively. The combination of the two methodologies in the cellular model showed intracellular concentrations of 0.344 ± 0.06 µg/mL and 2.483 ± 0.36 µg/mL for Zilpaterol and Clenbuterol, respectively. Extracellular concentration was 0.728 ± 0.14 µg/mL and 0.822 ± 0.11 µg/mL for Zilpaterol and Clenbuterol, respectively. This work shows the potential applications of cellular modelling in the study of toxicity for the mentioned drugs.
Assuntos
Clembuterol , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Células Hep G2 , Fígado , Espectrometria de Massas em Tandem , Compostos de TrimetilsililRESUMO
Perillyl alcohol (POH) is a monocyclic terpene that has strong antitumor activity. Brain tumors are particularly difficult to treat with therapeutic agents, and clinical trials have shown their low tolerance through oral administration. We proposed the entrapment of POH into an oil-in-water chitosan nanoemulsion aiming its intranasal administration for brain targeting. An ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantitation of total metabolite perillic acid (PA) in plasma and brain of rats. The rat samples containing the metabolite were treated by liquid-liquid extraction with acetonitrile. The mobile phase was 0.1% formic acid in water (solvent A) and 0.1% formic acid in methanol (solvent B), at a flow rate of 0.3 mL min-1 in gradient elution. The chromatography was run for 10 min, and analytical curves were built in acetonitrile, plasma, and brain. The PA was detected in positive ion mode with multiple reaction monitoring. The method has shown high selectivity, sensitivity, and throughput. The low quantification limits of 162, 178, and 121 ng mL-1 for acetonitrile, brain, and plasma, respectively, indicate a good detectability of the method. The repeatability and precision observed were within the limits recommended in the literature. The accuracy of the method was verified through high recovery rates (106-118%). The validated method was successfully applied to the pharmacokinetic study of the metabolite PA after the intranasal administration of free or POH-loaded nanoemulsion in rats. The results showed that chitosan nanoemulsion improved the plasma and brain bioavailability of POH, representing a promising alternative to free POH treatment.
Assuntos
Química Encefálica/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão/métodos , Cicloexenos , Emulsões , Monoterpenos , Administração Intranasal , Animais , Cicloexenos/análise , Cicloexenos/sangue , Cicloexenos/farmacocinética , Emulsões/administração & dosagem , Emulsões/química , Emulsões/farmacocinética , Limite de Detecção , Modelos Lineares , Monoterpenos/administração & dosagem , Monoterpenos/análise , Monoterpenos/sangue , Monoterpenos/química , Monoterpenos/farmacocinética , Nanoestruturas/administração & dosagem , Ratos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodosRESUMO
A quick and efficient method was optimized and validated to determine chlorpyrifos in biobeds using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). Chlorpyrifos was extracted from the matrix with 30 mL of a mixture of acetone, phosphoric acid and water 98:1:1 (v/v/v). After homogenization, centrifugation and filtration, 125 µL of the extract was evaporated and reconstituted in 5 mL of methanol acidified with 0.1% acetic acid. Validation was performed by studying analytical curve linearity (r2), estimated instrument and method limits of detection and limits of quantification (LODi, LODm, LOQi and LOQm, respectively), accuracy, precision (expressed as relative standard deviation, RSD), and matrix effect. Accuracy and precision were determined from the amount of pesticide recovered from biobed blank samples (i.e. without pesticide residue) spiked with chlorpyrifos at three different concentrations (2, 10 and 50 mg kg-1), with seven replicates at each concentration. For all three concentrations studied, the average recovery values obtained were between 96 and 115% with RSD values lower than 20%. The validated LOQ obtained was 2 mg kg-1 (from recovery studies) and the matrix effect observed was lower than ±20%, which demonstrated that there was neither considerable suppression nor enhancement of the analyte signal. The biobed system efficiently degraded chlorpyrifos in both 1) simulation of accidental spillage and 2) application of diluted pesticide solution. In the latter case, all the values obtained at the final sampling time (14 months) were below the validated LOQm.
Assuntos
Clorpirifos/análise , Cromatografia Líquida de Alta Pressão/métodos , Resíduos de Praguicidas/análise , Espectrometria de Massas em Tandem/métodos , Biodegradação Ambiental , Reatores Biológicos , Brasil , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos TestesRESUMO
Seafood represents a significant part of the human staple diet. In the recent years, the identification of emerging lipophilic marine toxins has increased, leading to the potential for consumers to be intoxicated by these toxins. In the present work, we investigate the presence of lipophilic marine toxins (both regulated and emerging) in commercial seafood products from non-European locations, including mussels Mytilus chilensis from Chile, clams Tawerea gayi and Metetrix lyrate from the Southeast Pacific and Vietnam, and food supplements based on mussels formulations of Perna canaliculus from New Zealand. All these products were purchased from European Union markets and they were analyzed by UPLC-MS/MS. Results showed the presence of the emerging pinnatoxin-G in mussels Mytilus chilensis at levels up to 5.2 µg/kg and azaspiracid-2 and pectenotoxin-2 in clams Tawera gayi up to 4.33 µg/kg and 10.88 µg/kg, respectively. This study confirms the presence of pinnatoxins in Chile, one of the major mussel producers worldwide. Chromatograms showed the presence of 13-desmethyl spirolide C in dietary supplements in the range of 33.2-97.9 µg/kg after an extraction with water and methanol from 0.39 g of the green lipped mussels powder. As far as we know, this constitutes the first time that an emerging cyclic imine toxin in dietary supplements is reported. Identifying new matrix, locations, and understanding emerging toxin distribution area are important for preventing the risks of spreading and contamination linked to these compounds.