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1.
Braz. arch. biol. technol ; 64: e21200163, 2021. tab, graf
Article in English | LILACS | ID: biblio-1153296

ABSTRACT

HIGHLIGHTS Isolate, fractionate and characterize extracts obtained from soursop leaves. Use of emerging green technologies such as microwave-ultrasound hybridization. The extracts contain kaempferol, procyanidins, catechin, and quercetin. The total ethanolic extract demonstrates cytotoxic effect on HeLa cells.


Abstract Cervical cancer is classified as the fourth most common malignancy in women. Natural compounds are a therapeutic alternative in cancer therapy. The aim of the study is to isolate, fractionate, and characterize extracts obtained from soursop leaves (Annona muricata L.) and determine their cytotoxic effect against HeLa cervical cancer cells and non-carcinogenic fibroblast 3T3 cells. The phytochemicals of soursop leaves were extracted through emerging green technologies such as the novel use of microwave-ultrasound hybridization and the use of environmentally friendly solvents (water and ethanol), in addition to the purification of extracts enriched in polyphenols by liquid chromatography with Amberlite XAD-16. Total aqueous and ethanolic extract were purified, as well as the fraction one of each extract. The extracts recovered from soursop leaves contained kaempferol and its isomers, procyanidins, catechin, and quercetin. The viability of the cells was determined with the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. HeLa and 3T3 cells were exposed to concentrations of 25, 50, 75, 100, 150, 200, and 250 ppm of a solution of soursop leaf extract powder. The MTT assay showed that soursop leaf extracts were toxic to both cell lines in general, however, the ethanolic extract at 25 and 50 ppm demonstrated inhibition in cell viability against the HeLa cancer line and low cytotoxicity for 3T3 fibroblast cells. In conclusion, the novel microwave-ultrasound hybridization technology allows the extraction of polyphenols that may have a potential cytotoxic effect on cancer cells.


Subject(s)
Humans , Female , HeLa Cells , Annona/chemistry , Polyphenols/isolation & purification , Phytochemicals/pharmacology , Antineoplastic Agents, Phytogenic/isolation & purification , Plant Extracts/pharmacology , Catechin/chemistry , Chromatography, Liquid/methods , Ethanol , Antineoplastic Agents, Phytogenic/pharmacology
2.
Article in English | WPRIM | ID: wpr-880319

ABSTRACT

BACKGROUND@#Glyphosate and its salt formulations are nonselective herbicides that have been extensively used worldwide, both for residential and agricultural purposes. The possible carcinogenicity and teratogenicity of glyphosate remain to be elucidated. We developed a sensitive and high-throughput analytical method for urinary glyphosate using liquid chromatography-tandem mass spectrometry with the aim of contributing to glyphosate exposure assessment in epidemiological studies.@*METHODS@#After urine dilution (creatinine matching dilution to 0.05 g creatinine/L), glyphosate was extracted using two types of solid phase extraction columns (SCX and NH2) with automated sample preparation instruments. The eluate was dried and dissolved in the mobile phase, followed by liquid chromatography-tandem mass spectrometry analysis. The optimized method was applied to urine samples obtained from 54 Japanese adults and children.@*RESULTS@#The results from the validation study demonstrated good recoveries (91.0-99.6%), within- and between-run precisions (< 15%), low detection limits (0.1 μg/L), and lower limit of quantification (0.3 μg/L). The detection frequency and median concentration of the urinary glyphosate in Japanese subjects were 59% and 0.25 μg/L (0.34 μg/g creatinine).@*CONCLUSIONS@#Our reliable determination method was successful in measuring urinary glyphosate concentration. Moreover, this is the first biomonitoring report of urinary glyphosate levels in the Japanese general population.


Subject(s)
Adult , Aged , Chromatography, Liquid/methods , Female , Glycine/urine , Humans , Male , Middle Aged , Solid Phase Extraction/methods , Tandem Mass Spectrometry/methods
3.
Arch. Clin. Psychiatry (Impr.) ; 46(5): 120-124, Sept.-Oct. 2019. tab, graf
Article in English | LILACS | ID: biblio-1054911

ABSTRACT

Abstract Background Current evidence suggests that upregulation of polyamines system plays a role both in cognitive deficit and synaptic loss observed in Alzheimer's disease (AD). Objective The aim of this study was to determine the plasmatic concentration of polyamines in mild cognitive impairment (MCI) and AD patients in comparison with healthy controls (HC). Methods Plasmatic polyamines were quantified using the AbsoluteIDQ® p180 and liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS). Results The study group comprised 34 AD patients, 20 MCI and 25 HC. All individuals were followed for 4 years. During this period 8 amnestic MCI patients (40% of the MCI sample at baseline) converted to AD. Spermidine level was lower in both patient groups (AD; MCI) compared to HC (p = 0.007). Plasma levels of spermine were higher in the MCI group (p < 0.001), but decreased in the sub-sample of MCI patients who converted to AD (p = 0.043). No statistically significant differences were found in ornithine and putrescine levels (p = 0.056 and p = 0.126, respectively). Discussion Our results suggest dynamic changes in the expression of polyamines in the MCI-AD continuum.


Subject(s)
Humans , Male , Female , Aged , Aged, 80 and over , Polyamines/blood , Spermine/blood , Alzheimer Disease/physiopathology , Cognitive Dysfunction/physiopathology , Ornithine/blood , Polyamines/metabolism , Biomarkers/blood , Putrescine/blood , Spermidine/blood , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Metabolomics/methods , Alzheimer Disease/diagnosis , Cognitive Dysfunction/diagnosis
4.
Rev. ciênc. farm. básica apl ; 4001/01/2019. ilus, tab
Article in English | LILACS | ID: biblio-1100195

ABSTRACT

Fenticonazole is an antifungal drug widely used in a cream formulation including as a generic medicine. Stability studies of fenticonazole in a cream formulation are very scarce. In this research, we intent to contribute to generic medicines quality control and provide reliable data seeking for insertion of fenticonazole monograph in official compendia. Therefore, in this work it was studied the behavior of fenticonazole under several conditions and developed a stability-indicating LC method to separate the degradation products and quantify the drug in presence of them, using the Design of Experiments (DoE) as tool to achieve robust and easy transferable method. Fenticonazole stability was evaluated under aqueous, alkaline (0.1 M NaOH), acidic (0.1 M HCL) and oxidative (3% v/v, H2O2) at ambient temperature and heating at 90°C, over 6 hours. The drug shows to be unstable under all stressed test conditions. It was completely degraded under acid medium with arising of degradation products. The robust and stability indicating LC method was validated. It is able to reveal the fenticonazole instability and to separate its degradation product with accuracy and precision (CV ˂ 2%) and without any placebo interferences.(AU)


Subject(s)
Humans , Chromatography, Liquid/methods , Imidazolines/analysis , Skin Cream/metabolism , Quality Control , Drug Stability
5.
São Paulo; s.n; s.n; 2019. 93 p. tab, ilus, graf.
Thesis in Portuguese, French | LILACS | ID: biblio-1025676

ABSTRACT

Os aldeídos são espécies reativas que podem ser produzidos endogenamente por processos como a lipoperoxidação, podendo reagir com lipídios, proteínas e DNA. Diversas evidências apontam para o envolvimento de aldeídos reativos na progressão de patologias como doenças cardiovasculares, arteriosclerose e doenças neurodegenerativas. Uma meta central do CEPIDRedoxoma é estudar a reatividade química de intermediários redox em ambientes biológicos e consequentes mudanças na estrutura e função de biomoléculas, entender como cada intermediário redox reage com biomoléculas específicas e os efeitos resultantes, essenciais para a concepção de biomarcadores e antioxidantes. O nosso grupo estuda os mecanismos de formação, detoxificação e reação com biomoléculas de aldeídos reativos endógenos e exógenos e seu papel em patologias como a esclerose lateral amiotrófica (ALS). Um dos mecanismos de detoxificação desses aldeídos é através da conjugação com a carnosina. Recentemente, foi observado que a suplementação de animais transgênicos ALS SOD G93A com carnosina via oral resultou em retardo da perda de peso e tendência de aumento da sobrevida dos animais. O presente projeto buscou investigar o possível papel da carnosina em animais modelo para ALS. Para isso as modificações em DNA induzidas por aldeídos reativos e a formação de adutos de carnosina-aldeídos foram analisadas através de metodologia HPLC-MS/MS. Assim observamos que ratos suplementados com carnosina apresentaram níveis significativamente menores de proteína carbonilada em músculo e fígado. Em fígadoforam vistos níveis menores de dois adutos de DNA, 8-oxodGuo e1,N2-HO-propanodGuo, em animais suplementados. Em cérebro foram detectados níveis menores de 1, N2-εdGuo. Com relação aos adutos carnosina-aldeídos, foi observado níveis significativamente maiores do aduto CAR-HHE na medula. Com embasamento nos resultados aqui apresentados, sugere-se a utilização de sequestradores de aldeídos como uma estratégia terapêutica em condições fisiopatológicas nas quais ao acúmulo dessas espécies está comprovado


Aldehydes are reactive species that can be produced endogenously by processes such as lipid peroxidation, which can react with lipids, proteins and DNA. Several evidences point to the involvement of reactive aldehydes in the progression of pathologies such as cardiovascular diseases, atherosclerosis and neurodegenerative diseases. A central goal of CEPID-Redoxoma is to study the chemical reactivity of redox intermediates in biological environments and consequent changes in the structure and function of biomolecules, to understand how each redox intermediate reacts with specific biomolecules and the resulting effects, essential for the design of biomarkers and antioxidants. Our group studies the mechanisms of formation, detoxification and reaction with biomolecules of endogenous and exogenous reactive aldehydes and their role in pathologies such as amyotrophic lateral sclerosis (ALS). One of the detoxification mechanisms of these aldehydes is through carnosine conjugation. Recently, we observed that oral carnosine supplementation in transgenic ALS SODG93A animals resulted in delayed weight loss and a tendency to increase the survival of the animals. The present project investigated the potential role of carnosine in animal models for ALS. Thus, reactive aldehydes induced DNA modifications and carnosine aldehyde adducts were analyzed by HPLC-MS/MS. We observed that rats supplemented with carnosine presented significantly lower levels of protein carbonylation in muscle and liver. Lower levels of two DNA adducts, 8-oxodGuo and 1, N2-HO-propanodGuo, were observed in liver of the supplemented animals. Lower levels of 1, N2-εdGuo were detected in the brain. Regarding the carnosine-aldehydeadducts, significantly higher levels of the CAR-HHE adduct were observed in spinal cord. The results presented here suggest the use of aldehyde scavengers as a therapeutic strategy under pathological conditions in which is proven the accumulation of these species


Subject(s)
Animals , Male , Female , Rats , Biological Phenomena , Carnosine/adverse effects , Aldehydes/analysis , Amyotrophic Lateral Sclerosis/pathology , Mass Spectrometry/methods , Chromatography, Liquid/methods , DNA Adducts
6.
Acta toxicol. argent ; 26(1): 19-31, mayo 2018. ilus, tab
Article in Spanish | LILACS | ID: biblio-973613

ABSTRACT

Los piretroides son insecticidas ampliamente usados no sólo en el ámbito agropecuario y doméstico sino también en salud pública. Una vez absorbidos, son rápidamente metabolizados a compuestos polares eliminados por vía renal. Uno de los metabolitos común a un gran número de piretroides es el ácido 3-fenoxibenzoico (3-PBA) el cual es utilizado como marcador de exposición. Se presenta en este trabajo, la validación de una metodología analítica para la determinación del 3-PBA utilizando QuEChERS acoplado a microextracción líquido-líquido dispersiva con tricloroetileno como disolvente extractivo y cromatografía líquida de alta resolución con detector de foto-arreglo de diodos. La validación se realizó con muestras aisladas de orina de voluntarios adultos de ambos sexos sin exposición conocida y orina sintética. El método resultó lineal en el intervalo 9 μg L-1-79 μg L-1; los límites de detección y cuantificación fueron de 3 μg L-1 y 9 μg L-1, respectivamente. No se observaron señales de interferentes a los tiempos de retención del 3-PBA y del ácido 2-fenoxibenzoico (2-PBA), estándar interno, en las muestras de orina blanco. Las señales cromatográficas en las muestras enriquecidas fueron espectralmente homogéneas. Las precisiones intradiarias (RSDr%) (n= 5) para 9 μg L-1 estuvieron comprendidas entre 9,3%-9,9% y para 27 μg L-1 entre 5,9%-10,6%. Las precisiciones interdiarias (RSDint%) (n=15) para los mismos niveles de concentración fueron de 11,8% y 9,1%, respectivamente. El rango de porcentajes de recuperación para 9 μg L-1 fue de 87%-119% y para 27 μg L-1 de 70%-91%. Se evaluó la estabilidad del analito en la muestra y en el extracto. El analito resultó estable a -20 °C durante 7 días en la muestra y durante 1 día en el extracto. Los valores de incertidumbre relativa e incertidumbre expandida fueron evaluados mediante la ecuación de Horwitz, los resultados obtenidos fueron para el nivel 9 μg L-1 de 33% y 65% y para el nivel 27 μg L-1 de 28% y 55%. La aplicabilidad del método validado fue evaluada con muestras reales de personas sin exposición laboral conocida, quienes declararon haber usado insecticidas piretroides. El método resultó sensible y selectivo.


Pyrethroid insecticides are used not only in the agricultural and domestic environment, but also in public health. Once absorbed, they are rapidly metabolized into polar compounds eliminated by the kidneys. One of the metabolites common to many pyrethroids is 3-phenoxybenzoic acid (3-PBA) which are used to evaluate exposure. We present in this paper the validation of an analytical methodology for the determination of 3-PBA using QuEChERS coupled to dispersive liquid-liquid microextraction with trichloroethylene as an extractive solvent and high-performance liquid chromatography with a photodiode array detector. Validation was carried out with isolated samples of urine from adult volunteers of both sexes without exposure and synthetic urine. The method was linear in the interval 9 μg L-1-79 μg L-1; the limit of detection and quantitation were 3 μg L-1 and 9 μg L-1, respectively. Interfering signals were not observed in the blank urine samples and the chromatographic signals in the enriched samples were spectrally homogeneous. The within-run precision (RSDr%) (n = 5) for 9 μg L-1 were between 9.3%-9.9% and for 27 μg L-1 between 5.9%-10.6%. The between-run precision (RSDint%) (n = 15) for the same concentration levels were 11.8% and 9.1%, respectively. The recovery for 9 μg L-1 ranged from 87%-119% and for 27 μg L-1 from 70%-91 %. The stability of the analyte was evaluated in the sample and in the extract. The analyte in the sample was stable at -20 °C for 7 days and in the extract was stable for 1 day. The values of relative uncertainty and expanded uncertainty obtained by the Horwitz equation were 33% and 65% for 9 μg L-1, and 28% and 55% for 27 μg L-1. The applicability of the validated method was evaluated with real samples of people without known occupational exposure, who declared having used pyrethroid insecticides. The method was sensitive and selective.


Subject(s)
Humans , Pyrethrins/poisoning , Pyrethrins/toxicity , Biomarkers/urine , Biomarkers/analysis , Chromatography, Liquid/methods , Insecticides/poisoning , Insecticides/toxicity
7.
Rev. cuba. obstet. ginecol ; 44(1): 1-11, ene.-mar. 2018.
Article in Spanish | LILACS | ID: biblio-978441

ABSTRACT

La macroprolactinemia se define como la presencia de cantidades elevadas de esta isoforma de la prolactina en suero, en conjunto con concentraciones normales de prolactina monomérica. Se trata de una entidad bastante común, considerada entre las tres primeras causas de hiperprolactinemia. Su origen parece responder a mecanismos autoinmunes y el seguimiento de los pacientes afectos durante periodos de 10 años ha demostrado que es una condición crónica. La prueba de elección para el diagnóstico es la cromatografía líquida en columna de gel, pero este es un método costoso que generalmente es suplido por la prueba de precipitación con polietinglicol. Por mucho tiempo ha prevalecido el concepto de que estos pacientes son esencialmente asintomáticos, pero reportes recientes señalan la presencia de síntomas de hiperprolactinemia como parte significativa del cuadro, aunque la literatura actual muestra criterios divergentes. En estos pacientes la realización de resonancia magnética nuclear hipofisaria parece ser un procedimiento innecesario, basado en la escasa frecuencia de resultados positivos. El tratamiento farmacológico con agonistas dopaminérgicos muestran respuestas contradictorias en cuanto a la desaparición de los síntomas y la normalización de los niveles de prolactina. Por tanto, la inexistencia de un consenso en la literatura científica en lo referente a las manifestaciones clínicas y el manejo, obliga a una conveniente valoración individual de cada caso(AU)


Macroprolactinemia is defined as the presence of high quantities of this prolactin isoform in serum, together with normal concentrations of monomeric prolactin. It is a common entity, considered among the three first causes of hyperprolactinaemia. The origin seems to respond to autoimmune mechanisms and the affected patients follow-up during ten years periods has shown that it is a chronic condition. The standard gold test to the diagnostic is gel-filtration chromatography, but it is an expensive method that is generally supplied by the polyethylene glycol precipitation test. During a long time, the concept that these patients are essentially asymptomatic has prevailed, but recent reports stamps the presence of hyperprolactinemia symptoms as a significant part of the entity, although current literature shows divergent criteria. In these patients, performing pituitary magnetic resonance seems to be an unnecessary procedure, based on the rare frequency of positive results. Pharmacological treatment with dopamine agonists shows contradictory responses with regard to symptoms disappearance and prolactin levels normalization. Therefore, the lack of consensus in the scientific literature with regard to the clinical manifestations and the management, requires a convenient individual assessment of each case(AU)


Subject(s)
Humans , Male , Female , Prolactin/analysis , Chromatography, Liquid/methods
8.
Rev. argent. endocrinol. metab ; 55(1): 43-56, mar. 2018. graf.
Article in Spanish | LILACS | ID: biblio-1248114

ABSTRACT

Esta revisión fue realizada con el fin de evaluar nuestros resultados de laboratorio así como aquellos de la literatura que constituyen, a nuestro entender, aportes significativos en el síndrome de ovarios poliquísticos (SOP). Nuestro especial énfasis será presentar las limitaciones de las metodologías empleadas por nuestro grupo, comparativamente a las reportadas por otros investigadores. La determinación de andrógenos, en particular de Testosterona (TT), es quizá la de mayor complejidad dado que los resultados con los diferentes inmunoensayos empleados en nuestro medio producen resultados muy variables por los diferentes métodos y aún entre laboratorios que usan la misma metodología. La técnica de referencia es la cromatografía líquida en tándem con espectrometría de masa (LC-MSMS), de difícil aplicación en laboratorios de análisis clínicos debido a su alto costo y la imposibilidad de resolver numerosas muestras. En estudios previos demostramos que de los métodos habitualmente usados para evaluar la TT circulante, solo en 2 inmunoensayos los resultados obtenidos fueron satisfactoriamente validados indirectamente según el criterio del Consenso de los Centros para el Control y Prevención de Enfermedades (CDC, USA) contra LC-MSMS, los cuales fueron comparables a dicha metodología con niveles superiores a 0,5 ng/ml. El SOP puede presentar factores de riesgo aumentados para la enfermedad cardiovascular y la diabetes II. Estos factores no están debidamente categorizados en función de los distintos fenotipos del SOP. Se evaluarán los principales analitos empleados con este objetivo y los nuevos que aporten elementos de mayor especificidad en este sentido


This review was performed in order to evaluate our laboratory results as well as those of the literature that constitute, in our opinion, significant contributions in these pathophysiologies. Our special emphasis will be on presenting the limitations of the methodologies used by our group, compared to those reported by other researchers. The determination of androgens, in particular Testosterone (TT), is perhaps the most complex since the results with the different immunoassays used in our environment produce very variable results by the different methods and even between laboratories that use the same methodology. The reference technique is LC-MSMS, difficult to apply in clinical analysis laboratories because of its high cost and the inability to solve numerous samples. In previous studies, we demonstrated that, in comparison to LC-MSMS with the usual methods for evaluating circulating TT, the results obtained in only 2 immunoassays were satisfactorily validated indirectly according to the criteria of CDC against LC-MSMS, which were comparable to that methodology with levels higher than 0.5 ng/ml. PCOS may have increased risk factors for cardiovascular disease and diabetes II. These factors are not properly categorized according to the different phenotypes of PCOS. The main analytes used for this purpose will be evaluated and new ones that contribute elements of greater specificity in this sense


Subject(s)
Humans , Female , Polycystic Ovary Syndrome/etiology , Polycystic Ovary Syndrome/physiopathology , Testosterone/analysis , Phenotype , Mass Spectrometry/methods , Immunoassay/methods , Chromatography, Liquid/methods
9.
São Paulo; s.n; s.n; 2018. 130 p. graf, tab, ilus.
Thesis in Portuguese | LILACS | ID: biblio-998555

ABSTRACT

A microextração por sorbente empacotado (MEPS) é uma técnica de preparo de amostras ainda pouco utilizada no âmbito da toxicologia, em que os mesmos princípios da extração em fase sólida convencional são adaptados para uma escala miniaturizada. As principais vantagens da técnica estão associadas ao pequeno volume de amostra e de solventes utilizados, à possibilidade de realizar múltiplas extrações com um mesmo cartucho e à facilidade de automação. Os benzodiazepínicos possuem grande relevância na toxicologia dada sua ampla utilização e seus efeitos que podem, por exemplo, comprometer a capacidade de dirigir, além do uso abusivo, e como drogas facilitadoras de crimes. Neste trabalho, um método de MEPS foi desenvolvido e otimizado para a determinação de sete benzodiazepínicos e seus produtos de biotransformação (diazepam, clonazepam, flunitrazepam, alprazolam, bromazepam, 7-aminoflunitrazepam e nordiazepam) utilizando 100 µL de amostra de sangue total post mortem. Após a extração, os eluatos foram analisados por cromatografia líquida em fase reversa acoplada a espectrometria de massas. O método foi validado de acordo com as recomendações do Scientific Working Group for Forensic Toxicology, apresentando linearidade adequada de 5 a 500 ng.mL-1 . Os valores de exatidão (90,4 a 109,5%), precisão intra-dia (2,5 a 10,7 %CV) e inter-dia (1,1 a 8,0 %CV) também foram satisfatórios. MEPS foi realizada mais de 60 vezes com a mesma fase extratora sem evidências de contaminação cruzada. Dez amostras reais fornecidas pelo Instituto Médico Legal de São Paulo foram analisadas. Foram quantificados diazepam, nordiazepam, clonazepam e bromazepam. Os resultados encontrados em cada uma das amostras foram comparados com dados da literatura


Microextraction by packed sorbent (MEPS) is a sample preparation technique still little used in toxicology, where the same principles of conventional solid phase extraction are adapted to a miniaturized scale. The main advantages of the technique are associated with the small volume of sample and solvents required, the possibility of performing multiple extractions with the same cartridge and ease process automation. Benzodiazepine drugs are relevant in toxicology because of their widespread use, and effects (which may, for example, compromise the ability to drive vehicles), abuse and records as crime-facilitating drugs. In this work, a MEPS method was developed and optimized for a determination of seven benzodiazepines and their metabolites (diazepam, nordiazepam, clonazepam, flunitrazepam, 7-aminoflunitrazepam, alprazolam, and bromazepam) using 100 µL of post mortem whole blood. After extraction, the eluates were analyzed by reversed-phase liquid chromatography coupled to mass spectrometry. The method was validated according to the recommendations of the Scientific Working Group for Forensic Toxicology, presenting adequate linearity from 5 to 500 ng.mL-1 . The values of accuracy (90.4 to 109.5%), intra-day precision (2.5 to 10.7 %CV) and inter-day (1.1 to 8.0 %CV) also presented satisfactory results. MEPS was performed more than 60 times with the same extractive phase without compromising the results with the evidence of carryover. Institute of Legal Medicine were submitted to analysis by MEPS-LC-MS/MS. In these samples, the following analytes were quantified: diazepam, nordiazepam, clonazepam and bromazepam. The results found in each of the samples were compared with data from the literature


Subject(s)
Benzodiazepines/analysis , Solid Phase Microextraction/instrumentation , Mass Spectrometry/methods , Autopsy , Computer Simulation/statistics & numerical data , Biotransformation , Chromatography, Liquid/methods , Drug Samples , Forensic Toxicology/classification
10.
Braz. J. Pharm. Sci. (Online) ; 54(2): e17491, 2018. tab, graf
Article in English | LILACS | ID: biblio-951933

ABSTRACT

ABSTRACT Multifunctional drug anisomycin was subjected to forced degradation in accordance with International Conference on Harmonisation (ICH) guidelines for the first time. The drug was exposed to the recommended stress conditions of hydrolysis (acidic, alkaline and neutral), oxidation, thermal stress and photolysis, in order to investigate its stability. Optimized LC-MS/MS method was validated as recommended by ICH Q2(R1) guideline with respect to the specificity, accuracy, precision, limits of detection and quantitation, linearity and robustness. Anisomycin exhibited high instability under alkaline and thermal (neutral hydrolysis) conditions. It showed moderate stability under acidic, neutral, oxidative, thermal (acidic hydrolysis) and photolytic conditions, with the lowest degradation level observed in the case of light and oxidation stress. Formation of the same degradation product, identified as deacetylanisomycin, was observed under all applied stress conditions.


Subject(s)
Evaluation Studies as Topic , Anisomycin/analysis , Mass Spectrometry/methods , Chromatography, Liquid/methods , Validation Study
11.
J. appl. oral sci ; 26: e20170561, 2018. tab
Article in English | LILACS | ID: biblio-954508

ABSTRACT

Abstract Saliva contains numerous proteins and peptides, each of them carries a number of biological functions that are very important in maintaining the oral cavity health and also yields information about both local and systemic diseases. Currently, proteomic analysis is the basis for large-scale identification of these proteins and discovery of new biomarkers for distinct diseases. Objective This study compared methodologies to extract salivary proteins for proteomic analysis. Material and Methods Saliva samples were collected from 10 healthy volunteers. In the first test, the necessity for using an albumin and IgG depletion column was evaluated, employing pooled samples from the 10 volunteers. In the second test, the analysis of the pooled samples was compared with individual analysis of one sample. Salivary proteins were extracted and processed for analysis by LC-ESI-MS/MS. Results In the first test, we identified only 35 proteins using the albumin and IgG depletion column, while we identified 248 proteins without using the column. In the second test, the pooled sample identified 212 proteins, such as carbonic anhydrase 6, cystatin isoforms, histatins 1 and 3, lysozyme C, mucin 7, protein S100A8 and S100A9, and statherin, while individual analysis identified 239 proteins, among which are carbonic anhydrase 6, cystatin isoforms, histatin 1 and 3, lactotransferrin, lyzozyme C, mucin 7, protein S100A8 and S100A9, serotransferrin, and statherin. Conclusions The standardization of protocol for salivary proteomic analysis was satisfactory, since the identification detected typical salivary proteins, among others. The results indicate that using the column for depletion of albumin and IgG is not necessary and that performing individual analysis of saliva samples is possible.


Subject(s)
Humans , Saliva/chemistry , Salivary Proteins and Peptides/analysis , Proteomics/methods , Proteomics/standards , Reference Standards , Immunoglobulin G , Reproducibility of Results , Chromatography, Liquid/methods , Albumins/analysis , Tandem Mass Spectrometry/methods
12.
Braz. J. Pharm. Sci. (Online) ; 54(1): e00188, 2018. tab, graf
Article in English | LILACS | ID: biblio-889443

ABSTRACT

ABSTRACT Fluoroquinolones are a known antibacterial class commonly used around the world. These compounds present relative stability and they may show some adverse effects according their distinct chemical structures. The chemical hydrolysis of five fluoroquinolones was studied using alkaline and photolytic degradation aiming to observe the differences in molecular reactivity. DFT/B3LYP-6.31G* was used to assist with understanding the chemical structure degradation. Gemifloxacin underwent degradation in alkaline medium. Gemifloxacin and danofloxacin showed more degradation perceptual indices in comparison with ciprofloxacin, enrofloxacin and norfloxacin in photolytic conditions. Some structural features were observed which may influence degradation, such as the presence of five member rings attached to the quinolone ring and the electrostatic positive charges, showed in maps of potential electrostatic charges. These measurements may be used in the design of effective and more stable fluoroquinolones as well as the investigation of degradation products from stress stability assays.


Subject(s)
Computer Simulation/statistics & numerical data , Fluoroquinolones/analysis , Fluoroquinolones/adverse effects , Ultraviolet Rays/adverse effects , Molecular Structure , Chromatography, Liquid/methods , Quinolones/analysis , Quinolones/chemistry
13.
Braz. J. Pharm. Sci. (Online) ; 54(1): e17381, 2018. tab, graf
Article in English | LILACS | ID: biblio-951900

ABSTRACT

Abstract A simple, sensitive, rapid and highly efficient LC-MS/MS method was developed for the determination of Candesartan and Hydrochlorothiazide simultaneously in human plasma. The method employed Zorbax eclipse C18 (150 X 4.6 mm, 5µ) column using acetate buffer: acetonitrile (25:75%, v/v) as the mobile phase. The mobile phase flow rate is 1 mL/min which was delivered into the mass spectrometer electron spray ionization chamber. The Liquid/liquid extraction procedure was used in the method for the extraction of analytes. The chromatograph was attached to a negative ion mode tandem mass spectrometer and the method was validated for all the parameters as per the guidelines of US-FDA. The ions were detected in multiple reaction monitoring mode and the transitions are m/z 439.00®309.10 and 295.80®268.80 for candesartan and hydrochlorothiazide respectively. Isotopic standards were used as internal standards for effective recovery of the analytes. The drugs were analyzed over a calibration range of 1.027-302.047 ng/mL for candesartan and 1.044-306.945 ng/mL for hydrochlorothiazide respectively with regression coefficient greater than 0.99. The mean extraction recoveries are 96.95±5.61 and 100.55±4.82 for candesartan and hydrochlorothiazide respectively. The precision and accuracy values for all the studies were within the range of ≤15% and 85-115%. The performed stability studies indicate that the developed method is stable in plasma for 15 h at room temperature (bench top); 52 h (in injector); for 112 days at -70 ºC for long term stability; five successive freeze and thaw cycles. The developed method could be successfully employed for the determination of selected drugs in biological samples.


Subject(s)
Plasma , Hydrochlorothiazide/analysis , Mass Spectrometry/methods , Chromatography, Liquid/methods , Validation Study
14.
Braz. J. Pharm. Sci. (Online) ; 54(2): e17163, 2018. tab, graf
Article in English | LILACS | ID: biblio-951946

ABSTRACT

ABSTRACT A simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of cabozantinib (CZ) in human plasma using cabozantinib-d4 (CZD4) as an internal standard (IS). Chromatographic separation was performed on Xbridge C18, 50 x 4.6 mm, 5 mm column with an isocratic mobile phase composed of 10mM Ammonium formate and Methanol in the ratio of (20:80 v/v), at a flow-rate of 0.7 mL/min. CZ and CZD4 were detected with proton adducts at m/z 502.2 ® 391.1 and 506.3 ® 391.2 in multiple reaction monitoring (MRM) positive mode respectively. Liquid-Liquid extraction method was used to extract the drug and IS. The method was validated over a linear concentration range of 5.0-5000.0 pg/mL with correlation coefficient (r2) ≥ 0.9994. This method demonstrated intra and inter-day precision within 1.95 to 2.37 and 2.93 to 9.3 % and Accuracy within 101.4 to 102.4 and 99.5 to 104.8 %. Cabozantinib was found to be stable throughout freeze-thawing cycles, bench top and postoperative stability studies


Subject(s)
Plasma , Pharmacokinetics , Validation Study , Protein-Tyrosine Kinases , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods
15.
Braz. J. Pharm. Sci. (Online) ; 54(4): e17242, 2018. tab, graf
Article in English | LILACS | ID: biblio-1001571

ABSTRACT

The novel alkaloid, oleracimine, presented remarkable anti-inflammatory bioactivity, and therefore, its pharmacokinetics was investigated in rat plasma after intravenous and oral administration by using a rapid ultra-high-performance liquid chromatography (UHPLC) method with UV detection at 270 nm. The analysis was performed on a shim-pack ODS column (75 mm×2 mm, 1.6 µm particle size, Shimadzu, Japan) column using isocratic elution with a mobile phase consisting of methanol-water (62:38, v/v) within 3 min. The results indicated that oleracimine was rapidly distributed with Tmax for 11.7 min after oral administration, which presented the double-peak phenomenon in the pharmacokinetic profile with a higher oral absolute bioavailability of 55.1% ± 7.83%.


Subject(s)
Animals , Male , Rats , Pharmacokinetics , Chromatography, Liquid/methods , Portulaca/adverse effects , Alkaloids/analysis
16.
São Paulo; s.n; s.n; 2018. 73 p. graf, tab.
Thesis in Portuguese | LILACS | ID: biblio-997407

ABSTRACT

As chamadas club drugs compreendem um vasto grupo de substâncias frequentemente utilizadas em bares, festas e raves, com a finalidade de intensificar o contato social e a estimulação sensorial. Englobam desde substâncias sintéticas comumente conhecidas, como a anfetamina, a metanfetamina, o MDMA, até moléculas de surgimento mais recente, denominadas novas substâncias psicoativas. Isoladas ou associadas a outras drogas, é possível que sejam causa de morte per se, ou que predisponham o usuário a envolver-se em situações potencialmente fatais, sendo necessário que os órgãos de Perícia Criminal (Institutos Médico Legais e Institutos de Criminalística) estejam aptos a detectar e quantificar essas substâncias em amostras biológicas. O presente trabalho teve como objetivo desenvolver um método analítico para identificação e quantificação de club drugs em sangue total, utilizando cromatografia líquida acoplada a espectrometria de massas com analisador híbrido quadrupolotempo de voo (LC-QTOF). Após o desenvolvimento do método, este foi validado utilizando as diretrizes do guia de validação do Scientific Working Group for Forensic Toxicology (SWGTOX), sendo analisados de linearidade, limite de detecção, limite de quantificação, efeito matriz, precisão intradia, precisão interdia, exatidão e integridade de diluição, além de recuperação e eficiência do processo. O método desenvolvido compreendeu a determinação de MDA, MDMA, 2C-B, DOB, cetamina, mCPP, cocaína e cocaetileno. Amostras provenientes de casos reais de morte não natural, oriundas do Instituto Médico Legal Aristoclides Teixeira de Goiânia - GO foram analisadas pelo método desenvolvido. 56 casos foram selecionados, em sua maioria com histórico de morte por projétil de arma de fogo e acidente de transito. Das 56 amostras analisadas, 28,5% (n=16) foram positivas para cocaína e/ou cocaetileno. As demais substâncias pesquisadas não foram encontradas nas amostras


Club drugs are a large group of substances consumed in pubs, parties and raves, aiming to intensify social contact and sensorial stimulation. The term comprises largely known substances such as amphetamine, methamphetamine, 3,4-methylenodioxymethamphetamine (MDMA), as well as so-called new psychoactive substances, which are synthetic drugs recently developed or recently introduced in drug market. Club drugs can be taken alone, combined with each other or, most frequently, with alcohol or other commonly abused drugs such as cocaine. In any of these situations, club drugs can possibly be the cause of death or potentialize the involvement of the user with crime and potentially fatal behavior. Thus, official organisms in charge of criminal investigation must be capable of identifying and quantifying these substances in biological samples. The present work aimed the development of an analytical method to identify and quantify club drugs in whole blood, using liquid chromatography - mass spectrometry with hybrid analyzer quadrupole - time of flight (LC-QTOF). After analytical development, the method was validated according to do Scientific Working Group for Forensic Toxicology (SWGTOX) guidelines, evaluating linearity, limit of detection, limit of quantification, matrix effect, precision, intermediate precision, bias and dilution integrity, besides recovery and process efficiency. The developed method comprised MDA, MDMA, 2C-B, DOB, ketamine, mCPP, cocaine and cocaethylene determination. Real samples related to non-natural deaths were collected at Institute of the Legal Medicine Aristoclides Teixeira, Goiânia, Goiás, Brazil, and analyzed by the developed method. 56 cases were selected, most of them related to fire gun injury and traffic events, 28,5% (n=16) of them being positive for cocaine and/or cocaethylene. None of the other drugs comprised in the analysis were detected in these samples


Subject(s)
Mass Spectrometry/methods , Illicit Drugs/analysis , Chromatography, Liquid/methods , Blood Chemical Analysis/statistics & numerical data , Blood Specimen Collection/statistics & numerical data , Substance-Related Disorders , Forensic Toxicology/instrumentation
17.
São Paulo; s.n; s.n; 2018. 87 p. tab, graf, ilus.
Thesis in Portuguese | LILACS | ID: biblio-997806

ABSTRACT

A inibição do quorum sensing (QS) altera a comunicação bacteriana, reduzindo a expressão de fatores de virulência e a formação de biofilmes, o que pode conferir menor pressão seletiva em comparação aos antibióticos tradicionais. As frutas e hortaliças constituem uma fonte rica em compostos com propriedades potenciais de inibição do QS. Entretanto, há pouca referência sobre o potencial de pimentas do gênero Capsicum e de seus compostos isolados como inibidores do QS. Esse trabalho teve como objetivo avaliar o efeito de extratos orgânicos obtidos das variedades de pimenta-malagueta e pimentão vermelho sobre o sistema QS dependente do sinalizador AI-1 (acil homoserina lactona - AHL) em bactérias Gram-negativas. Os extratos foram obtidos por extração em fase sólida e separados em uma fração metanólica e outra amônica; sendo os compostos característicos identificados e quantificados por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana dos extratos foi avaliada pela determinação da concentração inibitória mínima (MIC) e pela curva de crescimento de Chromobacterium violaceum ATCC 12472, Serratia liquefaciens MG1 e Pseudomonas aeruginosa PAO1. O efeito anti-QS dos extratos foi avaliado pelos testes de difusão em ágar e quantificação da produção de violaceína em meio líquido por C. violaceum e sobre a formação de biofilme, avaliado pelo ensaio de cristal violeta e microscopia confocal, em S. liquefaciens e P. aeruginosa nas temperaturas 30 ºC e 37 ºC. Os resultados obtidos pela CLAE indicaram que o extrato metanólico de pimenta-malagueta (EMPM) continha capsaicinoides como a capsaicina e dihidrocapsaicina, luteolina e outros compostos não identificados; já o extrato amônico desta não continha os compostos capsaicinoides. Ambos os extratos de pimentão vermelho continham luteolina e compostos não identificados, mas não apresentaram capsaicinoides. Como o EMPM era representativo dos demais extratos, por conter tanto capsaicinóides quanto luteolina, o foco deste trabalho foi avaliar os efeitos do EMPM sobre fenótipos microbianos nas concentrações 5; 2,5; 1,25 e 0,625 mg/ml, além de utilizar a capsaicina como controle comparativo em concentrações equivalentes às do extrato (25, 50 e 100 µg/ml). Os resultados da atividade antimicrobiana mostraram inibição parcial do crescimento das bactérias nas concentrações sub-MIC (MIC >5 mg/ml) de 5 e 2,5 mg/ml de EMPM. A capsaicina também inibiu parcialmente o crescimento das bactérias a 100 µg/ml, com exceção de S. liquefaciens a 37 ºC, cujo crescimento foi induzido em 50 e 25 µg/ml. A produção de violaceína foi reduzida pelo EMPM a 1,25 e 0,625 mg/ml, sem afetar o crescimento de C. violaceum. Ensaios com C. violaceum CV026, estirpe biosensora capaz de produzir o pigmento na presença de AI-1 exógeno, sugerem que o possível mecanismo de atuação do extrato sobre o sistema QS em C. violaceum 12472 é sobre a síntese do sinalizador, já que não foi observada inibição da produção de violaceína em CV026 pelo extrato. Contrariamente, a capsaicina incrementou a produção do pigmento na estirpe 12472, mas ensaios com a estirpe CV026 indicaram que a capsaicina não atua como sinalizador do QS, uma vez que esta não induziu a produção de violaceína nesta estirpe. Já a formação de biofilme foi incrementada na presença do EMPM, sendo consideravelmente maior em P. aeruginosa a 30 ºC. Igualmente, observou-se indução da formação de biofilme por capsaicina em S. liquefaciens (37 ºC) e P. aeruginosa (30 ºC). Porém, a capsaicina não teve efeito sobre a formação de biofilme de S. liquefaciens quando cultivada a 30 ºC, nem P. aeruginosa a 37 ºC. Os resultados revelam que a produção de violaceína em C. violaceum ATCC 12472 é inibida pelo EMPM, mas não pela capsaicina. Já, o EMPM e a capsaicina, de forma geral, não inibem a formação de biofilme de S. liquefaciens MG1 nem P. aeruginosa PAO1. Outros estudos são necessários para elucidar os mecanismos pelos quais o EMPM e a capsaicina agem sobre os fenótipos avaliados neste trabalho


Quorum sensing inhibition alters bacterial communication by reducing virulence factors expression and biofilm formation, exerting less selective pressure compared to antibiotics. Fruits and vegetables are rich sources of compounds with potential QS-inhibition properties. However, there are few references about the potential of peppers belonging to the genus Capsicum and its isolated compounds as QS inhibitors. This study aimed to assess the effect of organic extracts obtained from Capsicum varieties, pimenta-malagueta (red chili) and pimentão vermelho (red bell pepper), on the AI-1 dependent QS system. The extracts were obtained by solid phase extraction and split into a methanolic and an ammonic fraction. Characteristic compounds were identified and quantified by high performance liquid chromatography (HPLC). The antimicrobial activity of the extracts was assessed by determining the minimal inhibitory concentration (MIC) and the growth curve of Chromobacterium violaceum ATCC 12472, Serratia liquefaciens MG1 and Pseudomonas aeruginosa PAO1. The anti-QS effect of the extracts was evaluated by the agar diffusion assay and the quantification of violacein production was assessed in liquid medium by C. violaceum, as well as in the biofilm formation test determined by the crystal violet assay and confocal microscopy with S. liquefaciens and P. aeruginosa at 30 ºC and 37 ºC. HPLC results showed that the methanolic extract of pimenta-malagueta (EMPM) contained capsaicinoids such as capsaicin and dihidrocapsaicin, luteolin and other unidentified compounds in lower concentrations; while its ammonic extract did not have capsaicinoids. Both pimentão vermelho extracts contained luteolin and other unidentified compounds in low concentrations, but they did not contain capsaicinoids. As EMPM was representative among the extracts because it contained capsaicinoids and luteolin, the focus of this work was to assess the effect of EMPM over microbial phenotypes at concentrations of 5, 2.5, 1.25 and 0.625 mg/ml, using capsaicin as a comparative control at equivalent concentrations to those in EMPM (25, 50 and 100 µg/ml). Antimicrobial activity assays showed a partial inhibition growth of bacteria at sub-MIC concentrations (MIC >5 mg/ml) of EMPM at 5 and 2.5 mg/ml. Similarly, capsaicin partially inhibited bacterial growth at 100 µg/ml, except for S. liquefaciens at 37 ºC in which growth was induced at 50 and 25 µg/ml. Violacein production was reduced by EMPM at 1,25 and 0,625 mg/ml without affecting C. violaceum growth. Assays with C. violaceum CV026, a biosensor strain that produces violacein in the presence of exogenous AI-1, suggest that EMPM reduced violacein production in C. violaceum 12472 by interfering with the AI-1 synthesis. In contrast, capsaicin incremented violacein synthesis in strain 12472, but experiments with strain CV026 revealed that capsaicin does not function as an analog of AI-1. Biofilm formation was increased in EMPM presence, being remarkably superior in P. aeruginosa cultivated at 30 ºC, as opposed to cultivation at 37 ºC. Similarly, capsaicin induced biofilm formation in S. liquefaciens (37 ºC) and P. aeruginosa (30 ºC). However, capsaicin did not affect biofilm formation on S. liquefaciens cultured at 30 ºC, neither on P. aeruginosa at 37 ºC. These results show that violacein production in C. violaceum ATCC 12472 is inhibited by EMPM, but not by capsaicin. In general, EMPM and capsaicin did not inhibit biofilm formation in S. liquefaciens MG1 neither in P. aeruginosa PAO1. More studies are necessary to elucidate the mechanisms by which EMPM and capsaicin affect the studied phenotypes in this work


Subject(s)
Capsicum/adverse effects , Plant Extracts/analysis , Capsicum/adverse effects , Quorum Sensing , Gram-Negative Bacteria , Capsaicin/classification , Chromatography, Liquid/methods , Solid Phase Extraction/instrumentation
18.
Acta bioquím. clín. latinoam ; 51(4): 621-628, dic. 2017. ilus, graf, tab
Article in Spanish | LILACS | ID: biblio-886144

ABSTRACT

El cáncer de mama (CM) es una de las principales causas de muerte en México. Se ha observado un incremento en la incidencia de éste en mujeres de 15-29 años. A fin de comprender las causas en el desarrollo del CM, se pretendió buscar la asociación entre los genes/enfermedad empleando técnicas de Biología Molecular. Se analizaron, por genómica funcional, 50 biopsias frescas de pacientes con CM (BFCM), 50 biopsias embebidas en parafina de CM (BEPCM) y 10 biopsias frescas de pacientes con sospecha de CM (BFSC), obtenidas de mujeres que residen en Coahuila, México. Las muestras proteicas se cuantificaron y se resolvieron en geles de poliacrilamida dodecil sulfato de sodio (SDS-PAGE) y en dos dimensiones (2-DE). El perfil proteico de las BFCM, BEPCM versus BFSC mostró diferencias entre las bandas peptídicas observadas en los geles. Aquellos péptidos que se diferenciaron por su expresión fueron analizados por cromatografía líquida acoplada a masas en tándem (LC/ MS/MS). Las huellas peptídicas obtenidas, a su vez, se analizaron por medio del banco de genes (PubMed). Se encontraron, en las muestras de cáncer, proteínas asociadas a migración celular, supresión de tumores, estrés oxidativo y choque térmico. Por último, estos hallazgos se confirmaron empleando inmuno-electro transferencia o Western blot (WB) con anticuerpos contra vimentina.


Breast cancer (BC) is one of the leading causes of death in Mexico. Moreover, BC is the main cause of death in women between 15-29 years old in northern Mexico. Proteomic techniques have been used in order to achieve a better understanding of the genes involved in the development of BC. The proteins in BC extracted from 50 fresh breast cancer tissues (FBCT), 50 paraffin embedded breast cancer tissues (PEBCT) and 10 biopsies from women suspected of cancer (SC), residing in Coahuila, Mexico were analyzed in this paper. The quantity of protein extracted was similar in both samples FBCT and PEBCT. However, protein quality was lower in PEBCT than FBCT. Subsequently, these proteins were resolved in SDS-PAGE and 2DE. Differences were noticed in protein profile and all those suspect proteins were analyzed by LC/MS/MS. Amino acidic fingerprint allowed for the identification of peptides associated with a) cell migration, b) tumor suppression, c) oxidative stress or heat shock.


O câncer da mama (CM) é uma das principais causas de morte no México. Observou-se um aumento na incidência desse câncer em mulheres entre os 15-29 anos de idade. Para compreender as causas do desenvolvimento de CM, visou-se encontrar a associação entre os genes/doença utilizando técnicas de Biologia molecular. Analisaram-se por genômica funcional, 50 biópsias frescas de pacientes com CM (BFCM), 50 biópsias embebidas em parafina (BEPCM) e 10 biópsias frescas de pacientes com suspeita de CM (BFSC), obtidas de mulheres residentes em Coahuila, México. As amostras de proteínas foram quantificadas e separadas em géis de poliacrilamida dodecil sulfato de sódio (SDS-PAGE) e em duas dimensões (2-DE). O perfil proteico das BFCM, BEPCM comparado com BFSC mostrou diferenças entre as bandas peptídicas observadas nos géis. Esses peptídeos que diferem em sua expressão foram analisados por cromatografia líquida acoplada a massas em tandem (LC/MS/MS). As pegadas peptídicas obtidas, por sua vez, foram analisadas utilizando o banco de genes (PubMed). Verificaram-se nas amostras de câncer, proteínas associadas à migração celular, supressão de tumores, estresse oxidativo e choque térmico. Finalmente, estes achados foram confirmados utilizando a imuno-eletro transferência ou Western Blot (WB) com anticorpos contra vimentina.


Subject(s)
Humans , Female , Biomarkers/chemistry , Breast Neoplasms , Peptides/genetics , Chromatography, Liquid/methods , Mass Spectrometry/methods , Molecular Biology , Proteomics
19.
Acta toxicol. argent ; 24(1): 21-32, jul. 2016. graf, tab
Article in Spanish | LILACS | ID: biblio-837851

ABSTRACT

La Oficina de Naciones Unidas contra la Droga y el Delito (UNODC) en 2011 señala que "El delito facilitado por drogas (DFD) es una expresión general que abarca la violación y otras agresiones sexuales, el robo con violencia o intimidación, la extorsión de dinero y los malos tratos deliberados de ancianos o niños bajo la influencia de sustancias sicotrópicas". En este trabajo se validó un método cualitativo y rápido a partir de muestras de orina por LC/MS/MS para 39 compuestos comprendidos en los listados de sumisión química. El objetivo fue alcanzar un límite de detección un 50 % por debajo de la concentración propuesta como "Límites mínimos de funcionamiento exigidos (MRPL)" por la UNODC, para poder ser aplicado a muestras reales.


The United Nations Office on Drugs and Crime (UNODC) in 2011, states that "The Drug-facilitated crime (DFC) is a general term that includes rape or other sexual assault, robbery, money extortion, as well as the deliberate maltreatment of the elderly or children under the influence of psychotropic substances". In this work we validated a qualitative and fast method from urine samples by LC/MS/MS for 39 compounds included in the Drug-facilitated crime lists. The aim was to reach a detection limit of 50% below the proposed concentration as "minimum required performance limits (MRPL)" by UNODC in order to be applied in real samples.


Subject(s)
Humans , Chromatography, Gas/methods , Chromatography, Liquid/methods , Substance Abuse Detection/methods , Urine Specimen Collection/methods , Urine/chemistry , Sex Offenses , Substance-Related Disorders/urine , Urine Specimen Collection/statistics & numerical data
20.
Braz. j. pharm. sci ; 51(3): 533-540, July-Sept. 2015. tab, graf
Article in English | LILACS | ID: lil-766302

ABSTRACT

Peltodon longipes is used as a stimulant and emmenagogue. The objective of this study was to perform genotoxic and chromatographic analyses of the extracts of two samples of P. longipes, collected from the cities of Santa Maria and Tupanciretã, RS, Brazil. The Allium cepa assay was used to analyze genotoxicity while high-performance liquid chromatography was employed to determine phenolic compounds. The genotoxicity experiment consisted of nine groups each comprising four A. cepa bulbs. Bulb roots were developed in distilled water and then transferred for the treatments, for 24 hours, and the negative control remained in water. The treatments were: aqueous extracts at concentrations of 5 and 15 g L-1 for each sample, plus four groups treated with 1% glyphosate, one of which was used as a positive control and the other three for testing DNA damage recovery using water and the extracts of P. longipes from Santa Maria. All extracts of P. longipes exhibited anti-proliferative potential, although the effect was significantly greater for the extracts from the Tupanciretã sample. This sample also contained the highest amount of rosmarinic acid and kaempferol, which may confer the effects found in these extracts. Only extracts from the Santa Maria sample exhibited genotoxic potential.


Peltodon longipes é utilizada como estimulante e emenagoga. Objetivou-se realizar análises genotóxica e cromatográfica dos extratos de duas amostras de P. longipes, coletadas nos municípios de Santa Maria e Tupanciretã, RS, Brasil. O teste de Allium cepa foi utilizado para análise da genotoxicidade e a cromatografia líquida de alta eficiência, para determinação dos compostos fenólicos. O experimento de genotoxicidade constou de nove grupos de quatro bulbos de A. cepa. Os bulbos foram enraizados em água destilada e após transferidos para os tratamentos, por 24 horas, permanecendo o controle negativo em água. Os tratamentos foram: extratos aquosos nas concentrações de 5 e 15 g L-1 de cada amostra, além de quatro grupos tratados com glifosato 1%, um deles usado como controle positivo e outros três para testar a recuperação de danos ao DNA, utilizando água e os extratos de P. longipes da amostra de Santa Maria. Todos os extratos de P. longipes demonstraram potencial antiproliferativo, porém o efeito foi significativamente maior para os extratos da amostra de Tupanciretã. Essa amostra também apresentou maior quantidade de ácido rosmarínico e canferol, o que pode estar relacionado com os efeitos encontrados nesses extratos. Somente extratos da amostra de Santa Maria demonstraram potencial genotóxico.


Subject(s)
Genotoxicity/analysis , Lamiaceae/metabolism , Chromatography, Liquid/methods , Mentha/metabolism
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