RESUMO
A sensitive, selective, and rapid ultra-performance LC (UPLC)/MSIMS method was validated for the confirmation and quantification of clonazepam in human plasma. The analyte was extracted from human plasma with diethyl ether, reaching an average recovery of 64.02 and 66.48% for clonazepam and the internal standard, respectively. The separation was performed on a Waters ACQUITY UPLC BEH C18 column (50 x 2.1 mm id, 1.7 microm particle size) with gradient elution at a flow rate of 0.25 mL/min using a 0.5% formic acid solution (mobile phase A) and acetonitrile-methanol-formic acid (75+25 + 0.5, v/v/v; mobile phase B). Detection was performed on a triple-quadruple tandem mass spectrometer in the multiple reaction monitoring mode via electrospray ionization. Linear calibration curves were obtained in the concentration range of 0.3-50.0 ng/mL, with an LOQ of 0.3 ng/mL. The intraday and interday precision (CV) values were below 10%, and accuracy (relative error) ranged from -2.6 to 6.6% at all QC levels. The suggested method was successfully applied for the determination of clonazepam in human plasma in a bioequivalence study.
Assuntos
Cromatografia Líquida/métodos , Clonazepam/sangue , Psicotrópicos/sangue , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Calibragem , HumanosRESUMO
Introduction: Recent studies have suggested that cannabidiol (CBD) could interconvert into Delta-8- and Delta-9- tetrahydrocannabinol. Materials and Methods: Thus, we tested the plasma samples of 120 healthy human subjects (60 male and 60 female), 60 in fasting and the other 60 under normal feeding conditions after acute administration of an oral solution containing CBD 300 mg. To do this, we developed a bioanalytical method to determine CBD and the presence of THC in plasma samples by Ultra-High Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry. Results: The results showed that THC was not detected in plasma after the administration of CBD, and those study participants did not present psychotomimetic effects. Conclusions: The findings presented here are consistent with previous evidence suggesting that the oral administration of CBD in a corn oil formulation is a safe route for the administration of the active substance without bioconversion to THC in humans.
RESUMO
A sensitive and specific LC/MS/MS method was developed and validated for the determination of scopolamine butylbromide in human plasma. Scopolamine butylbromide and propanolol (internal standard) were extracted from the plasma by liquid-liquid extraction with dichloromethane as the extraction solvent and separated on a C18 analytical column (50 x 4.6 mm id) maintained at 40 degrees C. The analytes were eluted at a constant flow rate of 0.45 mL/min; the mobile phase consisted of acetonitrile and a buffer of 5 mM ammonium acetate and 0.1% formic acid (60 + 40, v/v). The mass spectrometer, equipped with an electrospray source in the positive ionization mode, was set up in the multiple-reaction monitoring mode to monitor the transitions m/z 360.6 > 102.5 (scopolamine butylbromide) and m/z 259.7 > 115.6 (propanolol). The chromatographic separation was obtained within 2.0 min, and the responses were linear over the concentration range of 0.10-40.00 ng/mL. The mean extraction recoveries of scopolamine butylbromide and propanolol from plasma were 69.00 and 80.76%, respectively. Method validation parameters, such as specificity, linearity, precision, accuracy, and stability, were within the acceptable range. Moreover, when the proposed method was successfully applied to a pharmacokinetic study of healthy human volunteers, the results showed that the two scopolamine butylbromide formulations tested are not bioequivalent in rate and extent of absorption.
Assuntos
Química Farmacêutica/métodos , Cromatografia Líquida/métodos , Hidrocarbonetos Bromados/análise , Hidrocarbonetos Bromados/sangue , Escopolamina/análise , Escopolamina/sangue , Espectrometria de Massas em Tandem/métodos , Acetatos/análise , Adolescente , Adulto , Técnicas de Química Analítica , Feminino , Formiatos/análise , Humanos , Masculino , Plasma/efeitos dos fármacos , Reprodutibilidade dos Testes , Solventes/química , Espectrometria de Massas por Ionização por Electrospray/métodos , TemperaturaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Vitex megapotamica (Spreng.) Moldenke is a deciduous tree, native of South America. Its leaves are traditionally used to treat cardiovascular diseases. This activity is related to the presence of flavonoids, the major compounds of the crude extract. AIM OF THE STUDY: This study investigated the effects of the oral administration of crude extract and standardized fractions from V. megapotamica leaves on lipid profile and on the formation of atherosclerotic plaque in C57BL/6 LDLr-KO mice treated with high-fat diet (HFD). MATERIALS AND METHODS: Male C57BL/6 LDLr-KO mice were fed with HFD (cholesterol, 1.25%) for 30 days. They were treated with hydroethanolic extract (500 or 1000mg/kg/day) or fractions (125 or 250mg/kg/day). After 30 days of treatment, it was evaluated the serum lipid profile, atherogenic index, and atherosclerotic plaque. RESULTS: All doses of the hydroethanolic extract reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index. The n-butanolic fraction also reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index, at all doses, with exception for the triglycerides, which only the lower dose was effective. The residual fraction reduced significantly the levels of total cholesterol, LDL-c and the atherogenic index, at all doses, with exception for the atherogenic index, which only the higher dose was effective. The atherosclerotic plaque formation was impaired only by the lower dose of the hydroethanolic extract. CONCLUSIONS: Overall, our data suggest that V. megapotamica has potential for the treatment of dyslipidemias.
Assuntos
Hiperlipidemias/induzido quimicamente , Hiperlipidemias/tratamento farmacológico , Hipolipemiantes/farmacologia , Extratos Vegetais/farmacologia , Vitex/química , Animais , Dieta Hiperlipídica/efeitos adversos , Hiperlipidemias/sangue , Hipolipemiantes/administração & dosagem , Lipídeos/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fitoterapia , Extratos Vegetais/química , Lectinas de Plantas/química , Receptores de LDL/genética , Receptores de LDL/metabolismoRESUMO
In the Myrtaceae family, the species Eugenia involucrata DC., popularly known as "cerejeira-do-mato", is traditionally used for the antidiarrheal and digestive action of its leaves. However, no studies were found in the literature regarding its antimicrobial and antioxidant potential. In this context, the objective of the present study was to determine the chemical composition by gas chromatography coupled to mass spectrometry (GC-MS) to evaluate the antimicrobial activity by the broth microdilution technique and the antioxidant activity by the 2,2-diphenyl-1-picryl-hydrazila (DPPH) method of the essential oil of E. involucrataleaves. GC-MS identified 28 compounds, all sesquiterpenes, corresponding to 89.41% of the essential oil. The antimicrobial activity of the essential oil was observed for all Gram-positive bacteria tested (Staplylococcus epidermidis, Enterococcus faecalis,Bacillus subtilis and Staplylococcus aureus) and for yeast Candida albicans. The essential oil presented a reduction capacity of DPPH up to 66.81%, evidencing its antioxidant potential. It is suggested that the antimicrobial and antioxidant action of E. involucrata essential oil is related to the presence of the major compounds, elixene (26.53%), ß-caryophyllene (13.16%), α-copaene (8.41%) and germacrene D (7.17%).
Na família Myrtaceae, a espécieEugenia involucrata DC. popularmente denominada "cerejeira-do-mato" é conhecida tradicionalmente pela ação antidiarreica e digestiva de suas folhas. Contudo, na literatura não foram encontrados trabalhos referentes ao seu potencial antimicrobiano e antioxidante. Neste contexto, o objetivo do presente estudo foi determinar a composição química por cromatografia gasosa acoplada a espectrometria de massas (CG-EM) e avaliar a atividade antimicrobiana pela técnica de microdiluição em caldo e a atividade antioxidante pelo método do 2,2-difenil-1-picril-hidrazila (DPPH) do óleo essencial das folhas de E. involucrata. A CG-EM identificou 28 compostos, todos sesquiterpenos, correspondendo a 89,41% do óleo essencial. A atividade antimicrobiana do óleo essencial foi observada para todas as bactérias Gram-positivas testadas (Staplylococcus epidermidis, Enterococcus faecalis,Bacillus subtilise Staplylococcus aureus) e para a levedura Candida albicans. O óleo essencial apresentou capacidade redutora de radicais DPPH de até 66,81%, evidenciando sua potencialidade antioxidante. Sugere-se que a ação antimicrobiana e antioxidante do óleo essencial de E. involucrata esteja relacionada à presença dos compostos majoritários, elixeno (26,53%), ß-cariofileno (13,16%), ï¡-copaeno (8,41%) e germacreno D (7,17%).
Assuntos
Myrtaceae , Eugenia , Anti-Infecciosos , Antioxidantes , Espectrometria de Massas , Bacillus , Bactérias , Produtos Biológicos , Candida albicans , Óleos , Destilação , Cromatografia Gasosa , Enterococcus faecalis , Compostos Orgânicos VoláteisRESUMO
A sensitive and fast ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) method for measurements of N-butylscopolamine in plasma was developed and validated. A single protein precipitation was proposed for the clean up of the plasma and N-methylhomatropine was added as internal standard (IS). The analyses were carried out using a C(18) column and mobile phase of acetonitrile: 5 mM ammonium acetate + 0.1% formic acid (90:10, v/v). The triple quadrupole mass spectrometer equipped with an electrospray source in positive mode, was set up in selective reaction monitoring, to detect precursor â product ion 360.0 â 194.0 m/z and 290.3 â 138.0 m/z transitions, for N-butylscopolamine and IS, respectively. The method was linear in 0.03 (lower limit of quantitation; LLOQ) - 10.00 ng/ml range for N-butylscopolamine. Satisfactory selectivity, linearity, precision, accuracy, and robustness were obtained for the UPLC-ESI-MS/MS method. The proposed method was successfully applied to a pharmacokinetic study of healthy human volunteers; the results showed that the two scopolamine butylbromide formulations tested are not bioequivalent in rate and extent of absorption.
Assuntos
Brometo de Butilescopolamônio/sangue , Antagonistas Muscarínicos/sangue , Espectrometria de Massas por Ionização por Electrospray/métodos , Adulto , Cromatografia Líquida de Alta Pressão/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Espectrometria de Massas em Tandem/métodos , Adulto JovemRESUMO
The purpose of this study is to compare the bioavailability of two itraconazole (CAS 84625-61-6) capsule formulations. An open, randomized, two-period crossover study with a 7-day washout interval was conduced in 32 healthy volunteers. The plasma samples were obtained up to 96 h after drug administration. A sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of itraconazole in human plasma. Itraconazole and ketoconazole (internal standard) were extracted from the plasma by liquid-liquid extraction using diethylether : dichloromethane (70 : 30) as extraction solvent and separated on a C8 analytical column (150 mm x 4.6 mm I.D.) maintained at 40 degrees C. The elution was performed by a constant flow rate of 1.2 mL/min and the mobile phase consisted of acetonitrile and acetic acid 0.1% (85 :15 v/v). The mass spectrometer equipped with an electrospray source in positive mode, was set up in multiple reaction monitoring, to detect parent --> production 705.0 392.0 (itraconazole) and 531.0 --> 81.70 (ketoconazole). The chromatographic separation was obtained within 3.5 min and was linear in the concentration range of 5 to 600 ng/mL. Bioequivalence between the products was determined by calculating 90% confidence intervals for the ratio of C(max) (95.02%-109.48%), AUC(0-t) (81.41%-107.77%) and AUC(0-inf) (80.85%-106.86%). These values for the test and reference products are within the 80-125% interval, proposed by FDA and EMEA. It was concluded that the proposed method was successfully applied to a pharmacokinetic study in healthy human volunteers, and results showed that the two itraconazole formulations are bioequivalent in their rate and extent of absorption.