RESUMO
Endoplasmic reticulum (ER) stress is a cellular state that results from the overload of unfolded/misfolded protein in the ER that, if not resolved properly, can lead to cell death. Both acute lung infections and chronic lung diseases have been found related to ER stress. Yet no study has been presented integrating metabolomic and transcriptomic data from total lung in interpreting the pathogenic state of ER stress. Total mouse lungs were used to perform LC-MS and RNA sequencing in relevance to ER stress. Untargeted metabolomics revealed 16 metabolites of aberrant levels with statistical significance while transcriptomics revealed 1593 genes abnormally expressed. Enrichment results demonstrated the injury ER stress inflicted upon lung through the alteration of multiple critical pathways involving energy expenditure, signal transduction, and redox homeostasis. Ultimately, we have presented p-cresol sulfate (PCS) and trimethylamine N-oxide (TMAO) as two potential ER stress biomarkers. Glutathione metabolism stood out in both omics as a notably altered pathway that believed to take important roles in maintaining the redox homeostasis in the cells critical for the development and relief of ER stress, in consistence with the existing reports.
Assuntos
Cresóis/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Glutationa/metabolismo , Lesão Pulmonar/diagnóstico , Metilaminas/metabolismo , Ésteres do Ácido Sulfúrico/metabolismo , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Cresóis/análise , Estresse do Retículo Endoplasmático/genética , Perfilação da Expressão Gênica/métodos , Pulmão/química , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/genética , Lesão Pulmonar/metabolismo , Lesão Pulmonar/patologia , Masculino , Metabolômica/métodos , Metilaminas/análise , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/genética , Estresse Oxidativo/fisiologia , Ésteres do Ácido Sulfúrico/análise , Resposta a Proteínas não Dobradas/genética , Resposta a Proteínas não Dobradas/fisiologiaRESUMO
Alcohol and tobacco are the most frequently consumed substances in the world. Both are significantly associated with the increasing number of different diseases. Thus, monitoring nicotine and alcohol use is vital for public health planning and intervention strategies. This study aimed to calculate estimates of alcohol and nicotine use in 11 cities of Turkey using wastewater-based epidemiology. In 2019, daily composite wastewater samples from 18 wastewater treatment plants were collected for a week per season. The 24-h composite samples were collected via auto-samplers. Sample preparation for wastewater samples collected was done using liquid-liquid extraction and solid-phase extraction. Nicotine and ethyl sulfate (EtS) were analyzed using validated liquid chromatography-tandem mass spectrometry method. The estimated average nicotine consumption was 2.84 mg/p/day, and the average alcohol consumption was 3.46 ± 1.83 ml/p/day. The highest nicotine consumption was observed in Kayseri city; the highest alcohol consumption was calculated for Mersin city. In this study, the cigarette and alcohol consumption estimate obtained by wastewater-based epidemiology (WBE) was found to be higher than the Turkey Tobacco and Alcohol Market Regulatory Authority report. To our knowledge, this study is the most comprehensive one so far applied using WBE for 11 cities in Turkey and evaluates alcohol and nicotine use together.
Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia , Uso de Tabaco/epidemiologia , Vigilância Epidemiológica Baseada em Águas Residuárias , Águas Residuárias/análise , Cromatografia Líquida/métodos , Cidades , Humanos , Extração Líquido-Líquido , Nicotina/análise , Estações do Ano , Extração em Fase Sólida , Ésteres do Ácido Sulfúrico/análise , Espectrometria de Massas em Tandem/métodos , Turquia/epidemiologiaRESUMO
Wastewater-based epidemiology (WBE), an emerging tool for monitoring public health in near real-time, is used extensively in Europe but applications to U.S. populations are still scarce. In this longitudinal study, raw wastewater was collected monthly from three U.S. cities as 24-h weekday composites and analyzed for evidence of alcohol and tobacco consumption. Over the 11-month sampling period, biomarkers of stimulant use were detected in wastewater by isotope dilution liquid chromatography tandem mass spectrometry in units of µg/L (ethyl sulfate, 1.6-25.1; nicotine, 0.6-26.7; cotinine, 0.2-3.8; and 3hydroxycotinine, 0.3-3.8). Average consumption rates in the three communities were calculated using detected biomarker levels in conjunction with wastewater flow rates, metabolic excretion factors, and population size data. Computed average per-capita consumption rates estimated for the sub-population aged 15 and above for alcohol (13.4⯱â¯5.6â¯L/y/person) and daily consumption of nicotine by smokers (14.2⯱â¯3.6 cigarettes/d/person) were in good agreement with U.S. survey data (9.0â¯L/y/person; 14.2 cigarettes/d/smoker). The WBE approach also captured impacts of temporal population influx on substance consumption patterns. This first U.S. WBE study to track recreational use of stimulants longitudinally and concurrently in multiple American cities highlights opportunities for collecting robust public health information from wastewater anonymously, economically and in near real-time.
Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia , Águas Residuárias/análise , Consumo de Bebidas Alcoólicas/tendências , Cromatografia Líquida , Cotinina/análogos & derivados , Cotinina/análise , Humanos , Meio-Oeste dos Estados Unidos/epidemiologia , New England/epidemiologia , Nicotina/análise , Estações do Ano , Fumar/epidemiologia , Fumar/tendências , Ésteres do Ácido Sulfúrico/análise , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Diethyl sulfate (DES) is a substance classified to the group of carcinogens. The value of maximum admissible concentration for this substance in workplace air is not specified in Poland. Due to the use of DES in domestic companies there is a need to develop a sensitive method for the determination of diethyl sulfate in the work environment. MATERIAL AND METHODS: Studies were performed using gas chromatography (GC) technique. An Agilent Technologies chromatograph, series 7890A, with a mass selective detector (5975C, Agilent Technologies, USA) was used in the experiment. Separation was performed on a capillary column with Rtx-5MS (30 m × 0.25 mm × 0.25 µm) (Restek, USA). The possibility of using sorbent tubes filled with activated carbon (100 mg/50 mg), silica gel (100 mg/50 mg) and Porapak Q (150 mg/75 mg) for absorption of diethyl sulphate was investigated. RESULTS: The method of sampling air containing diethyl sulfate was developed. Among the sorbents to absorb DES Porapak Q was chosen. Determination of the adsorbed vapor includes desorption of DES, using dichloromethane/methanol mixture (95:5, v/v) and chromatographic analysis of so obtained solution. Method is linear (r = 0.999) within the investigated working range of 0.27- -5.42 µg/ml, which is an equivalent to air concentrations 0.0075-0.15 mg/m3 for a 36 l air sample. CONCLUSIONS: The analytical method described in this paper allows for selective determination of diethyl sulfate in the workplace air in the presence of dimethyl sulfate, ethanol, dichloromethane, triethylamine, 2-(diethylamino)ethanol, and triethylenetetramine. The method meets the criteria for performing procedures aimed at measuring chemical agents, listed in EN 482. Med Pr 2018;69(3):291-300.
Assuntos
Poluentes Ocupacionais do Ar/análise , Monitoramento Ambiental/métodos , Exposição por Inalação/análise , Exposição Ocupacional/análise , Ésteres do Ácido Sulfúrico/análise , Cromatografia Gasosa/métodos , Humanos , Material Particulado/análise , Polônia , Ésteres do Ácido Sulfúrico/administração & dosagem , Ésteres do Ácido Sulfúrico/efeitos adversos , Local de TrabalhoRESUMO
Since alcohol and tobacco consumption are among the leading causes of population health harm, it is very important to understand the consumption behaviour to develop effective harm reduction strategies. Wastewater-based epidemiology (WBE) is a potential tool for estimating their consumption, but there are several uncertainties that need to be determined, including the stability of biomarkers in the sewer. Utilizing a real rising main sewer, this study investigated the stability of alcohol and tobacco consumption biomarkers. Rhodamine and acesulfame were used as flow tracer and benchmarker to understand the transportation of wastewater in the sewer with a hydraulic retention time between 2.7 and 5.0â¯h. Ethyl sulphate (EtS) and ethyl glucuronide (EtG), two biomarkers of alcohol consumption, were found to have different in-sewer stability, with EtS much more stable than EtG. The degradation rate of EtS is approximately 8% per hour, while EtG has a half-life of 1.9â¯h. Formation of nicotine, cotinine and trans-3'-hydroxycotinine, three biomarkers for tobacco consumption, was observed during the experiment, probably due to deconjugation of their glucuronide chemicals. The deconjugation process has prevented the determination of actual stability of the three chemicals. However, it is suggested that cotinine is relatively stable, while nicotine and trans-3'-hydroxycotinine degrade to a certain degree in the sewer system. According to our findings, the in-sewer degradation is more important during the interpretation of alcohol consumption estimation than for tobacco consumption estimation.
Assuntos
Consumo de Bebidas Alcoólicas , Uso de Tabaco , Águas Residuárias/análise , Poluentes Químicos da Água/análise , Biomarcadores/análise , Cotinina/análogos & derivados , Cotinina/análise , Etanol/metabolismo , Glucuronatos/análise , Humanos , Nicotina/análise , Ésteres do Ácido Sulfúrico/análiseRESUMO
Understanding the actual consumption of alcohol and tobacco in the population is important for forming public health policy. For this purpose, wastewater-based epidemiology has been applied as a complementary method to estimate the overall alcohol and tobacco consumption in different communities. However, the stability of their consumption biomarkers - ethyl sulfate, ethyl glucuronide, cotinine, and trans-3'-hydroxycotinine - in the sewer system has not yet been assessed. This study aimed to conduct such assessment using sewer reactors mimicking conditions of rising main, gravity sewer, and wastewater alone, over a 12-hour period. The results show that cotinine and trans-3'-hydroxycotinine are relatively stable under all sewer conditions while ethyl sulfate was only stable in wastewater alone and gradually degraded in rising main and gravity sewer conditions. Ethyl glucuronide quickly degraded in all reactors. These findings suggest that cotinine and trans-3'-hydroxycotinine are good biomarkers to estimate tobacco consumption; ethyl sulfate may be used as a biomarker to estimate alcohol consumption, but its in-sewer loss should be accounted for in the calculation of consumption estimates. Ethyl glucuronide, and probably most of glucuronide compounds, are not suitable biomarkers to be used in wastewater-based epidemiology due to their in-sewer instability.
Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia , Cotinina/análogos & derivados , Glucuronatos/análise , Esgotos/análise , Ésteres do Ácido Sulfúrico/análise , Uso de Tabaco/epidemiologia , Poluentes Químicos da Água/análise , Biomarcadores/análise , Cotinina/análise , Etanol/análise , Humanos , Nicotiana/química , Águas Residuárias/análiseRESUMO
Wastewater analysis has been demonstrated to be a complementary approach for assessing the overall patterns of drug use by a population while the full potential of wastewater-based epidemiology has yet to be explored. F2-isoprostanes are a prototype wastewater biomarker to study the cumulative oxidative stress at a community level. In this work, 8-iso-prostaglandin F2α (8-iso-PGF2α) was analysed in raw 24 h-composite wastewater samples collected from 4 Norwegian and 7 other European cities in 2014 and 2015. Using the same samples, biomarkers of alcohol (ethyl sulfate) and tobacco (trans-3'-hydroxycotinine) use were also analysed to investigate any possible correlation between 8-iso-PGF2α and the consumption of the two drugs. The estimated per capita daily loads of 8-iso-PGF2α in the 11 cities ranged between 2.5 and 9.9 mg/day/1000 inhabitants with a population-weighted mean of 4.8 mg/day/1000 inhabitants. There were no temporal trends observed in the levels of 8-iso-PGF2α, however, spatial differences were found at the inter-city level correlating to the degree of urbanisation. The 8-iso-PGF2α mass load was found to be strongly associated with that of trans-3'-hydroxycotinine while it showed no correlation with ethyl sulfate. The present study shows the potential for 8-iso-PGF2α as a wastewater biomarker for the assessment of community public health.
Assuntos
Cotinina/análogos & derivados , Dinoprosta/análogos & derivados , Uso de Tabaco/epidemiologia , Águas Residuárias/análise , Biomarcadores/análise , Cidades , Cotinina/análise , Dinoprosta/análise , Europa (Continente) , Humanos , Noruega , Estresse Oxidativo , Análise de Regressão , Ésteres do Ácido Sulfúrico/análise , UrbanizaçãoRESUMO
Alcohol consumption in young women is a widespread habit that may continue during pregnancy and induce alterations in the fetus. We aimed to characterize prevalence of alcohol consumption in parturient women and to assess fetal ethanol exposure in their newborns by analyzing two direct metabolites of ethanol in meconium. This is a cross-sectional study performed in September 2011 and March 2012 in a series of women admitted to an obstetric unit following childbirth. During admission, socio-demographic and substance use (alcohol, tobacco, cannabis, cocaine, and opiates) during pregnancy were assessed using a structured questionnaire and clinical charts. We also recorded the characteristics of pregnancy, childbirth, and neonates. The meconium analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect the presence of ethyl glucuronide (EtG) and ethyl sulfate (EtS). Fifty-one parturient and 52 neonates were included and 48 meconium samples were suitable for EtG and EtS detection. The median age of women was 30 years (interquartile range (IQR): 26-34 years); EtG was present in all meconium samples and median concentration of EtG was 67.9 ng/g (IQR: 36.0-110.6 ng/g). With respect to EtS, it was undetectable (<0.01 ng/g) in the majority of samples (79.1%). Only three (6%) women reported alcohol consumption during pregnancy in face-to-face interviews. However, prevalence of fetal exposure to alcohol through the detection of EtG and EtS was 4.2% and 16.7%, respectively. Prevention of alcohol consumption during pregnancy and the detection of substance use with markers of fetal exposure are essential components of maternal and child health.
Assuntos
Consumo de Bebidas Alcoólicas/metabolismo , Etanol/metabolismo , Troca Materno-Fetal , Mecônio/química , Adulto , Consumo de Bebidas Alcoólicas/efeitos adversos , Feminino , Glucuronatos/análise , Humanos , Gravidez , Ésteres do Ácido Sulfúrico/análiseRESUMO
BACKGROUND/AIMS: Alcohol and nicotine are the two most used substances world-wide and associated with increased burden of disease. Since surveys on substance use may be difficult due to response biases, wastewater-based epidemiology (WBE) was developed as a more objective measure of nicotine and alcohol use. This study compares estimates of nicotine and alcohol use from a wastewater sampling campaign in a medium-sized Belgian city with a concurrently executed population survey. METHODS: 29,083 letters about participation in an online survey study on weekly alcohol and tobacco use were sent to the inhabitants of Lier, Belgium. Wastewater samples were collected from the associated treatment plant in four bi-weekly periods. Samples were analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Ethylsulfate was used as alcohol biomarker and cotinine and trans-3'-hydroxycotinine as nicotine biomarker. RESULTS: 263 (1%) surveys were filled out on average per week. According to survey data, alcohol and nicotine were used less than in the rest of Belgium and this was matched by the wastewater data. Nicotine use, but not alcohol use, showed a significant variation over the sampling periods. Both nicotine and alcohol showed increase use during the weekend while only alcohol showed a different use pattern throughout the week. CONCLUSION: No correlation between WBE and survey data could be demonstrated, possibly due to small sample sizes. However, this study shows that weekly trends in alcohol and nicotine use can be quickly detected from wastewater analysis and the occurrence of major events such as festivals can be identified.
Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia , Etanol/análise , Nicotina/análise , Fumar/epidemiologia , Águas Residuárias/análise , Adolescente , Adulto , Idoso , Bélgica/epidemiologia , Biomarcadores/análise , Cromatografia Líquida/métodos , Cidades/epidemiologia , Cotinina/análogos & derivados , Cotinina/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Características de Residência , Ésteres do Ácido Sulfúrico/análise , Inquéritos e Questionários , Espectrometria de Massas em Tandem , Adulto JovemRESUMO
During a routine determination of dimethyl sulfate in technical materials using gas chromatography-mass spectrometry (GC-MS), we found that residual monomethyl sulfate originating from a prior methylation reaction with dimethyl sulfate decomposed in the hot GC injection system to yield dimethyl sulfate and sulfuric acid. This thermal reaction leads to false positive or overestimated residue levels of dimethyl sulfate, accompanied by bad chromatographic peak shapes and poor precision and accuracy values. This short communication describes proper measures to counteract this problem and presents a fast, reliable and validated GC-MS method that is capable of determining dimethyl sulfate residues in the presence of monomethyl sulfate in technical materials using a simple dissolve-and-inject approach. Applying deuterated dimethyl sulfate as internal standard and with a sample weight of 25 mg, a limit of detection of 0.24 mg kg(-1) and a limit of quantification of 0.48 mg kg(-1) was achieved along with a linear range of 0.48-208.6 mg kg(-1). The method offers a precision of 9.1% and an accuracy of 96.5% at the limit of quantification and a precision of 3.6% and an accuracy of 93.8% at a dimethyl sulfate level of 1 mg kg(-1).
Assuntos
Carcinógenos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ésteres do Ácido Sulfúrico/análise , Limite de DetecçãoRESUMO
(-)-Epicatechin, an abundant dietary polyphenol found mainly in cocoa and tea, is known to extensively undergo metabolism after ingestion giving rise to a complex series of conjugated metabolites including numerous isomers. In the present study, the combination of fractionation, chemical derivatization and various mass spectrometric approaches is described to determine the exact position of sulphate group in methylated epicatechin metabolites. Four O-methyl-(-)-epicatechin-O-sulphate metabolites isolated from human urine samples were derivatized under mild condition using trimethylsilyldiazomethane (TMSD) in the presence of methanol. The resulting methylated reaction products were then analyzed by high resolution and multistage mass spectrometry for the subsequent identification of the sulphate positional isomers. Results show that O-methylation affects the charge delocalization in negatively charged ions and hereby the fragmentation pattern of the sulphate isomers allowing the identification of diagnostic ions. In addition, this study demonstrates that methoxy derivatives of polyphenol metabolites can be prepared using TMSD. Subsequently, the localization of the sulphate group in the polyphenol metabolites can be achieved by analyzing the methoxy derivatives by multistage mass spectrometry. Using an enzymatic reaction for identification of the O-methyl position, and a chemical O-methylation with TMSD follow by high resolution and multistage tandem MS for the identification of the sulphate group position, we were able to identify the previously unknown O-methyl-(-)-epicatechin-O-sulphate. Accordingly, we identified 3'-O-methyl-(-)-epicatechin-5-O-sulphate and 3'-O-methyl-(-)-epicatechin-7-O-sulphate as the main O-methyl-(-)-epicatechin-sulfates(-)-epicatechin metabolites in humans.
Assuntos
Catequina/análogos & derivados , Espectrometria de Massas/métodos , Ésteres do Ácido Sulfúrico/análise , Catequina/análise , Catequina/metabolismo , Catequina/urina , Diazometano/análogos & derivados , Diazometano/química , Humanos , Ésteres do Ácido Sulfúrico/metabolismo , Ésteres do Ácido Sulfúrico/urina , Compostos de Trimetilsilil/químicaRESUMO
A simple and rapid capillary zone electrophoretic method was developed for determining dimethyl sulfate a possible human carcinogen and mutagen and chloroacetyl chloride a potential genotoxic agent at trace levels in pharmaceutical drug substances by indirect photometric detection. A systematic screening of various anionic probes was performed to obtain the best separation conditions and sensitivity. High sensitivities with low quantification and detection levels were achieved for dimethylsulfate and chloroacetyl chloride using a background electrolyte (BGE) containing 5 mM pyridine dicarboxylic acid as the probe ion. The method is specific, precise and accurate for the two genotoxins. The optimized method was validated for specificity, precision, linearity, accuracy and stability in solution. Calibration curves were linear (R>0.999) for both dimethylsulfate and chloroacetyl chloride in the range LOQ - 300% of nominal concentrations. The CE method was effectively implemented for estimating dimethylsulfate and chloroacetyl chloride in two different active pharmaceutical ingredients (APIs).
Assuntos
Acetatos/análise , Carcinógenos/análise , Eletroforese Capilar/métodos , Mutagênicos/análise , Preparações Farmacêuticas/análise , Ésteres do Ácido Sulfúrico/análise , Acetatos/química , Calibragem , Carcinógenos/química , Eletrólitos/química , Mutagênicos/química , Preparações Farmacêuticas/química , Sensibilidade e Especificidade , Ésteres do Ácido Sulfúrico/químicaRESUMO
Toad venom, namely, "Chansu" in China, has been widely used for the treatment of heart failure, sores, pains, and various cancers. Upon LC-MS analysis of the venom from Bufo melanosticus collected in Indonesia, new bufadienolide sulfates were identified. For a complete characterization, the MeOH extract of the toad venom from B. melanosticus was fractionated by preparative HPLC, and the structures of five new buadienolide sulfates (1-5) along with one new bufogenin (6) were rapidly elucidated on the basis of LC-DAD-MS(n) and LC-SPE-NMR data. The in vitro growth inhibitory activity of these six compounds along with hellebrin (positive control) has been assayed by means of the MTT colorimetric assay in four human and two mouse cancer cell lines. Compound 3 and hellebrin displayed similar and marked in vitro cytotoxicity.
Assuntos
Venenos de Anfíbios/química , Antineoplásicos , Bufanolídeos , Bufonidae , Ésteres do Ácido Sulfúrico , Animais , Antineoplásicos/análise , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Bufanolídeos/análise , Bufanolídeos/química , Bufanolídeos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Indonésia , Camundongos , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Ésteres do Ácido Sulfúrico/análise , Ésteres do Ácido Sulfúrico/química , Ésteres do Ácido Sulfúrico/farmacologiaRESUMO
Deferasirox (ICL670) is a novel once-daily, orally administered iron chelator to treat chronic iron overload in patients with transfusion-dependent anemias. Absorption, distribution, metabolism, and excretion of [14C]deferasirox at pharmacokinetic steady state was investigated in five adult beta-thalassemic patients. Deferasirox (1000 mg) was given orally once daily for 6 days to achieve steady state. On day 7, patients received a single oral 1000-mg dose (approximately 20 mg/kg) of [14C]deferasirox (2.5 MBq). Blood, plasma, feces, and urine samples collected over 7 days were analyzed for radioactivity, deferasirox, its iron complex Fe-[deferasirox]2, and metabolites. Deferasirox was well absorbed. Deferasirox and its iron complex accounted for 87 and 10%, respectively, of the radioactivity in plasma (area under the curve at steady state). Excretion occurred largely in the feces (84% of dose), and 60% of the radioactivity in the feces was identified as deferasirox. Apparently unchanged deferasirox in feces was partly attributable to incomplete intestinal absorption and partly to hepatobiliary elimination of deferasirox (including first-pass elimination) and of its glucuronide. Renal excretion was only 8% of the dose and included mainly the glucuronide M6. Oxidative metabolism by cytochrome 450 enzymes to M1 [5-hydroxy (OH) deferasirox, presumably by CYP1A] and M4 (5'-OH deferasirox, by CYP2D6) was minor (6 and 2% of the dose, respectively). Direct and indirect evidence indicates that the main pathway of deferasirox metabolism is via glucuronidation to metabolites M3 (acyl glucuronide) and M6 (2-O-glucuronide).
Assuntos
Benzoatos/metabolismo , Benzoatos/farmacocinética , Sobrecarga de Ferro/tratamento farmacológico , Reação Transfusional , Triazóis/metabolismo , Triazóis/farmacocinética , Talassemia beta/terapia , Adulto , Animais , Área Sob a Curva , Arilsulfatases/metabolismo , Benzoatos/efeitos adversos , Benzoatos/uso terapêutico , Células Cultivadas , Deferasirox , Fezes/química , Feminino , Glucuronidase/metabolismo , Glucuronídeos/análise , Glucuronídeos/metabolismo , Hepatócitos/enzimologia , Humanos , Hidroxilação , Quelantes de Ferro/efeitos adversos , Quelantes de Ferro/metabolismo , Quelantes de Ferro/farmacocinética , Quelantes de Ferro/uso terapêutico , Masculino , Estrutura Molecular , Ratos , Espectrometria de Massas por Ionização por Electrospray , Ésteres do Ácido Sulfúrico/análise , Ésteres do Ácido Sulfúrico/metabolismo , Triazóis/efeitos adversos , Triazóis/uso terapêutico , Adulto JovemRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The aerial parts of Sisymbrium officinale Scop. are commonly used to treat airway ailments, moreover in antiquity the herbal drug was reputed to possess anticancer properties. The results obtained in present work support the traditional use and the properties ascribed to Sisymbrium officinale. AIM OF THE STUDY: In order to give a scientific basis to the traditional uses of Sisymbrium officinale, this study was aimed to evaluate in vitro the myorelaxant activity, the antimicrobial properties and the antimutagenic effect of an aqueous dry extract of the aerial parts of the plant. A phytochemical characterization of the extract was also performed. MATERIALS AND METHODS: The myorelaxant activity was studied against the contractions induced by carbachol, histamine and leukotriene C(4), in isolated guinea-pig trachea. The antimicrobial activity was tested against six bacteria and one yeast. The Ames test, performed by the preincubation method, was used to study the antimutagenic activity of the extract by its capability to inhibit the mutagenic effect of 2-nitrofluorene, sodium azide, methyl methanesulfonate and 2-aminoanthracene, in Salmonella typhimurium TA98, Salmonella typhimurium TA100 and Escherichia coli WP2uvrA strains. The chemical composition of the extract was analyzed by TLC and HPLC. RESULTS: Sisymbrium officinale showed to reduce the chemically-induced contractions of isolated guinea-pig trachea with major potency against leukotriene C(4) and histamine. The extract did not show any antibacterial activity. The Ames test showed a strong antimutagenic activity against 2-aminoanthracene, in Escherichia coli WP2uvrA and in Salmonella typhimurium TA98 strains. The phytochemical study highlighted the presence of putranjivine, the glucosinolate marker of Sisymbrium officinale, and of proline. CONCLUSIONS: The myorelaxant activity of Sisymbrium officinale offers a scientific basis to its use in traditional medicine. The strong antimutagenic effect suggests further studies to evaluate its possible chemopreventive activity.
Assuntos
Antimutagênicos/farmacologia , Brassicaceae/química , Glucosinolatos/análise , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Parassimpatolíticos/farmacologia , Extratos Vegetais/farmacologia , Ésteres do Ácido Sulfúrico/análise , Animais , Antracenos , Antimutagênicos/análise , Bactérias/efeitos dos fármacos , Glucosinolatos/farmacologia , Cobaias , Histamina , Leucotrieno C4 , Parassimpatolíticos/análise , Componentes Aéreos da Planta , Extratos Vegetais/química , Prolina/análise , Prolina/farmacologia , Ésteres do Ácido Sulfúrico/farmacologia , Traqueia , Leveduras/efeitos dos fármacosRESUMO
Dimethyl sulfate (DMS) is an alkylating reagent commonly used in organic syntheses and pharmaceutical manufacturing processes. Due to its potential carcinogenicity, the level of DMS in the API process needs to be carefully monitored. However, in-process testing for DMS is challenging because of its reactivity and polarity as well as complex matrix effects. In this short communication, we report a GC-MS method for determination of DMS in an API intermediate that is a methyl sulfate salt. To overcome the complex matrix interference, DMS and an internal standard, d6-DMS, were extracted from the matrix with methyl tert-butyl ether. GC separation was conducted on a DB-624 column (30 m long, 0.32 mm ID, 1.8 microm film thickness). MS detection was performed on a single-quad Agilent MSD equipped with an electron impact source while the MSD signal was acquired in selected ion monitoring mode. This GC/MS method showed a linear response for DMS equivalent from 1.0 to 60 ppm. The practical quantitation limit for DMS was 1.0 ppm and the practical detection limit was 0.3 ppm. The relative standard derivation for analyte response was found as 0.1% for six injections of a working standard equivalent to 18.6 ppm of DMS. The spike recovery was ranged from 102.1 to 108.5% for a sample of API intermediate spiked with 8.0 ppm of DMS. In summary, the GC/MS method showed adequate specificity, linearity, sensitivity, repeatability and accuracy for determination of DMS in the API intermediate. This method has been successfully applied to study the efficiency of removing DMS from the process.
Assuntos
Química Farmacêutica/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ésteres do Ácido Sulfúrico/análise , Ésteres do Ácido Sulfúrico/química , Química/métodos , Estabilidade de Medicamentos , Hidrólise , Íons , Modelos Químicos , Padrões de Referência , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização por Electrospray/métodos , Água/químicaRESUMO
OBJECTIVE: To develop a headspace gas chromatography method for determining dimethyl sulphate residual in granisetron hydrochloride. METHODS: An Angilent INNOWAX capillary column with nitrogen gas as carrier and FID as detector was applied in this study. Dimethyl sulphate was tested under a constant column temperature. RESULT: Dimethyl sulphate had different retention time from other organic solvents such as alcohol,acetoacetate, isopropanol, dichlormethane and chloroform, which might exist in granisetron hydrochloride. The detection limit of dimethyl sulphate;s was 0.0016%. CONCLUSION: The method can be used for the determination of dimethyl sulphate residual in granisetron hydrochloride.
Assuntos
Cromatografia Gasosa , Contaminação de Medicamentos , Granisetron/análise , Ésteres do Ácido Sulfúrico/análise , Cromatografia Gasosa/métodos , Contaminação de Medicamentos/prevenção & controle , Preparações Farmacêuticas/análiseRESUMO
Alkyl sulfates (AS), alkyl ethoxysulfates (AES) and linear alkyl benzene sulfonates (LAS) are all High Production Volume (HPV) and 'down-the-drain' chemicals used globally in detergent and personal care products, resulting in low levels ultimately released to the environment via wastewater effluent. Due to their surfactant properties, they preferentially sorb to sediments. Hence, assessment of their levels and potential perturbations on benthos are of interest. The relative levels of AS/AES decreased with distance from the wastewater treatment plant outfall. However, this was not evident for LAS. Short chained AES and especially AS dominated the homologue distribution for AES. There were no evident patterns in LAS homologue distribution. The overall mean margin of exposure (MoE) for AS/AES and LAS is approximately 40 (range: 3 to 100) suggesting no noteworthy perturbation on biota. The findings in this study are in concordance with previous preliminary hazard screening. Comparative sediment contamination analyses principally based on Chapman and Anderson [Chapman PM, Anderson, J. A decision-making framework for sediment contamination. Integr Environ Assess Mana. 2005; 1: 163-173.] and the U.S. Environmental Protection Agency RAPID assessment methods [USEPA. Rapid bioassessment protocols for use in wadeable streams and rivers: Periphyton, benthic, macroinvertebrates, and fish. 1999. Second Edition. U.S. Environmental Protection Agency Office of Water, Washington, D.C. EPA 841-B-99-002.] did not reveal significant correlations between the surfactant concentrations and ecological status of the sampling locations. Several Lines of Evidence (LoE) of the Weight-of-Evidence (WoE) lead to the conclusion of low aquatic risk associated to the monitored compounds.
Assuntos
Ácidos Alcanossulfônicos/análise , Ésteres do Ácido Sulfúrico/análise , Tensoativos/análise , Eliminação de Resíduos Líquidos , Poluentes Químicos da Água/análise , Ácidos Alcanossulfônicos/toxicidade , Animais , Biodiversidade , Monitoramento Ambiental , Sedimentos Geológicos/análise , Indiana , Invertebrados , Nível de Efeito Adverso não Observado , Ohio , Densidade Demográfica , Relação Quantitativa Estrutura-Atividade , Medição de Risco , Rios , Ésteres do Ácido Sulfúrico/toxicidade , Tensoativos/toxicidade , Poluentes Químicos da Água/toxicidadeRESUMO
This study reports the discovery of a novel sulfonated oxysterol found at high levels in the mitochondria and nuclei of primary rat hepatocytes after overexpression of the gene encoding steroidogenic acute regulatory protein (StarD1). Forty-eight hours after infection of primary rat hepatocytes with recombinant adenovirus encoding StarD1, rates of bile acid synthesis increased by 4-fold. Concurrently, [(14)C]cholesterol metabolites (oxysterols) were increased dramatically in both the mitochondria and nuclei of StarD1-overexpressing cells, but not in culture medium. A water-soluble [(14)C]oxysterol product was isolated and purified by chemical extraction and reverse-phase HPLC. Enzymatic digestion, HPLC, and tandem mass spectrometry analysis identified the water-soluble oxysterol as 5-cholesten-3beta,25-diol 3-sulfonate. Further experiments detected this cholesterol metabolite in the nuclei of normal human liver tissues. Based upon these observations, we hypothesized a new pathway by which cholesterol is metabolized in the mitochondrion.
Assuntos
Hepatócitos/química , Hidroxicolesteróis/análise , Ésteres do Ácido Sulfúrico/análise , Animais , Núcleo Celular/química , Células Cultivadas , Colesterol/metabolismo , Colesterol 7-alfa-Hidroxilase/biossíntese , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida/métodos , Humanos , Espectrometria de Massas/métodos , Mitocôndrias Hepáticas/química , Fosfoproteínas/biossíntese , RatosRESUMO
Alkyl sulfates (AS) and alkyl ethoxysulfates (AES) are High Production Volume (HPV) 'down-the-drain' chemicals widely used globally in detergent and personal care products, resulting in low levels (ng to microg L(-1) range) ultimately released to the environment via wastewater. These surfactants have a strong affinity for sorption to sediments. However, data regarding the fate and effects following release into the environment has not been reported. Sediment samples from both normal exposed and presumably low exposed locations (background) were analyzed to determine the levels of AS/AES. The method used in this study shows broad applicability across various sediment types and the most common congeners of AS/AES. The combined levels of AS/AES detected in the two presumed lower exposed sites ranged from 0.025 and 0.034 microg g(-1) on a dry weight (dw) basis while the presumed higher exposed site had combined levels of AS/AES of 0.117 microg g(-1) (dw) based on triplicate analyses. Results indicate that detectable levels of AS/AES can be found in sediments in the environment at these three sites that are below the concentrations expected to produce significant adverse ecological effects for individual homologues and the whole mixture, the hazard screening for these three sites had PEC(porewater)/PNEC(total mixture) ratios of 0.007-0.024. However, further investigation of potential effects and risk assessment is warranted.