RESUMEN
The current study presents a convenient, rapid and effective simultaneous extraction/derivatization (SEDP) strategy for effective pretreatment of catecholamines (CAs). Commercial zirconium oxide (ZrO2) nanoparticles were employed for the selective capturing of cis-diol containing CAs to remove the biological interferences and phenyl isothiocyanate (PITC) was used for derivatization to improve the ionization and to improve the chromatographic separation. The extraction and derivatization procedures were integrated into one step to simplify the sample pretreatment. Excessive derivatization reagents were removed as well, reducing the degree of contaminations in mass spectrometry. The factors affecting the SEDP process were optimized and the results showed that the detection sensitivity and chromatographic separation of CAs greatly improved compared with underivatized CAs, during LC-MS/MS analysis. Combined with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), quantifying the concentration of norepinephrine (NE), epinephrine (E) and dopamine (DA) in biological fluids was validated in ranges of 1-200.0 ng/mL with a satisfactory correlation coefficient (R2 > 0.997). The obtained recoveries were in the range of 91.0-109.5% with RSDs less than 9.4%. Finally, significant changes in CAs levels in urine samples of healthy people and pheochromocytoma patients were detected. The developed method offers comparative advantages in terms of sensitivity, specificity and selectivity.
Asunto(s)
Neoplasias de las Glándulas Suprarrenales , Catecolaminas , Cromatografía Liquida , Feocromocitoma , Espectrometría de Masas en Tándem , Urinálisis , Catecolaminas/análisis , Catecolaminas/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Humanos , Feocromocitoma/orina , Sensibilidad y Especificidad , Extracción en Fase Sólida , Urinálisis/métodosRESUMEN
Tobacco-specific N-nitrosamines are carcinogenic components in mainstream cigarette smoke. To explore tobacco-specific N-nitrosamine release levels in cigarettes, a magnetic solid-phase extraction procedure using magnetic graphene composite as sorbent for fast enrichment of tobacco-specific N-nitrosamine was developed. Under optimal conditions, a tobacco-specific N-nitrosamine determination method was successfully proposed by combining magnetic solid-phase extraction procedure and high-performance liquid chromatography coupled with tandem mass spectrometry. The method's limit of detection for tobacco-specific N-nitrosamines in mainstream cigarette smoke ranged from 0.018 to 0.057 ng/cigarette. Good linearities were obtained with correlation coefficients above 0.9992. The accuracies of tobacco-specific N-nitrosamines in a spiked mainstream cigarette smoke sample were from 89.3 to 109.4%, with a relative standard deviation of less than 11.2%. The proposed method has the merits of rapidity and high sensitivity. Finally, the method was successfully applied to tobacco-specific N-nitrosamine analysis in real samples.
Asunto(s)
Grafito/química , Nicotiana/química , Nitrosaminas/análisis , Extracción en Fase Sólida , Adsorción , Fenómenos Magnéticos , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
In this study, a silica/primary secondary amine (SiO2/PSA) was used as an in-pipette-tip solid phase extraction (SPE) sorbent for the simultaneous determination of polycyclic aromatic hydrocarbons (PAHs) and tobacco-specific N-nitrosamines (TSNAs) in mainstream cigarette smoke (MSS). We investigated several parameters including an extraction procedure of total particulate matter, type and amount of sorbent and on-line gel permeation chromatography parameters to obtain optimum conditions for a new strategy to target analytes. Under the optimized conditions, we developed a method for the simultaneous determination of PAHs and TSNAs in MSS by coupling in-pipette-tip SPE procedures to an on-line gel permeation chromatography-gas chromatography-tandem mass spectrometry (on-line GPC-GC-MS(2)). Our method had limits of detection for target analytes ranging from 0.01 to 0.23ng/cig. Good linearities were obtained with coefficients of determination (R(2)) greater than 0.9984 for all target analytes. Good reproducibility was obtained as intra- and inter-day precisions, and the relative standard deviations were less than 11.4 and 13.3%, respectively. The recoveries were in the range of 77.1-108.6% at different concentrations for real samples. Compared to previous standard methods for the determination of PAHs and TSNAs in MSS, our method was highly effective, fast, and had low consumption of organic solvent and a high degree of automation. Finally, our method successfully analyzed PAHs and TSNAs in real samples, and no significant deviations were observed when compared to similar analysis using standard methods.
Asunto(s)
Técnicas de Química Analítica/métodos , Cromatografía en Gel , Cromatografía de Gases y Espectrometría de Masas , Nitrosaminas/análisis , Hidrocarburos Policíclicos Aromáticos/análisis , Humo/análisis , Extracción en Fase Sólida , Técnicas de Química Analítica/normas , Límite de Detección , Reproducibilidad de los Resultados , Dióxido de Silicio/química , Espectrometría de Masas en Tándem , Nicotiana/químicaRESUMEN
In this study, magnetic graphene was used as modified quick, easy, cheap, effective, rugged and safe (QuEChERS) adsorbent for the determination of organochlorine pesticide (OCPs) residues in tobacco. To achieve the optimum conditions of modified QuEChERS procedure toward target analytes, several parameters affecting the clean-up efficiency including the amount of the adsorbent and clean-up time were investigated. Under the optimized conditions, a method for the determination of 26 OCPs residues in tobacco was established by coupling the modified QuEChERS procedure to on-line gel permeation chromatography-gas chromatography-tandem mass spectrometry (on-line GPC-GC-MS(2)). The limits of detection of proposed method for 26 OCPs residues ranged from 0.01275 to 3.150ng/g. And good linearities of the proposed method were obtained with coefficients of determination (R(2)) greater than 0.9985 for all target analytes. Good reproducibility of method was obtained as intra- and inter-day precisions, the relative standard deviations were less than 11.1 and 15.0%, respectively. The apparent recoveries were in the range of 64-126% at different concentrations for real samples. Compared with the reported methods for the determination of OCPs residues in tobacco, the proposed method has the advantages of simple to operate, low cost and high clean-up ability. Finally, the method was successfully applied to the analysis of OCPs residues in real samples.
Asunto(s)
Técnicas de Química Analítica/métodos , Contaminantes Ambientales/análisis , Grafito/química , Fenómenos Magnéticos , Nicotiana/química , Residuos de Plaguicidas/análisis , Técnicas de Química Analítica/economía , Cromatografía en Gel , Cromatografía de Gases y Espectrometría de Masas , Hidrocarburos Clorados/análisis , Reproducibilidad de los ResultadosRESUMEN
In this study, a cytotoxicity assay was developed for profiling the cytotoxicity of cigarette smoke condensates (CSCs) base on a cellular impedance biosensor (CIB). Compared with the traditional in vitro cytotoxicity assays, this CIB-based method offered distinct advantages in real-time kinetic measurement which provided a comprehensive understanding of cellular responses for the entire duration of the experiment and prediction of the potential mechanism of action of a given treatment. The time-dependent cell response profiles provided valid evidences for optimization of cell number per well, cell quality control, and identification of the optimal time points for compound treatment and endpoint assays. According to the time dependent IC50 values, the CIB could provide dynamic information that can be used to identify maximum toxicity of cigarette smoke and reversibility of the toxic effects which are difficult to achieve by the endpoint assays. The comparative IC50 values indicated that the as-developed biosensor offered analytical results in good consistency with the commonly used NRU method. The features of the CIB-based cytotoxicity assay, such as no cell labeling, automatic detection, and easy operation, give this assay potential to become routine setting for evaluating the cytotoxicity of CSCs.