Simultaneous determination of dextromethorphan, dextrorphan and doxylamine in human plasma by HPLC coupled to electrospray ionization tandem mass spectrometry: Application to a pharmacokinetic study

In the present study, a fast, sensitive and robust method to quantify dextromethorphan, dextrorphan and doxylamine in human plasma using deuterated internal standards (IS) is described. The analytes and the IS were extracted from plasma by a liquid-liquid extraction (LLE) using diethyl-ether/hexane (80/20, v/v). Extracted samples were analyzed by high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Chromatographic separation was performed by pumping the mobile phase (acetonitrile/water/formic acid (90/9/1, v/v/v) during 4.0 min at a flow-rate of 1.5 mL min(-1) into a Phenomenex Gemini (R) C18, 5 mu m analytical column (150 x 4.6 mm id.). The calibration curve was linear over the range from 0.2 to 200 ng mL(-1) for dextromethorphan and doxylamine and 0.05 to 10 ng mL(-1) for dextrorphan. The intra-batch precision and accuracy (%CV) of the method ranged from 2.5 to 9.5%, and 88.9 to 105.1%, respectively. Method inter-batch precision (%CV) and accuracy ranged from 6.7 to 10.3%, and 92.2 to 107.1%, respectively. The run-time was for 4 min. The analytical procedure herein described was used to assess the pharmacokinetics of dextromethorphan, dextrorphan and doxylamine in healthy volunteers after a single oral dose of a formulation containing 30 mg of dextromethorphan hydrobromide and 12.5 mg of doxylamine succinate. The method has high sensitivity, specificity and allows high throughput analysis required for a pharmacokinetic study. (C) 2012 Elsevier B.V. All rights reserved.

A Phase I clinical trial of lodenafil carbonate, a new phosphodiesterase Type 5 (PDE5) inhibitor, in healthy male volunteers

Lodenafil carbonate is a new phosphodiesterase Type 5 (PDE5) inhibitor used in treatment of erectile dysfunction. Objective: The present study was conducted to evaluate the safety, tolerability, and pharmacokinetics of lodenafil carbonate after administering ascending (1 - 100 mg) single oral doses to healthy male volunteers (n = 33). Methods: The study was an open-label, dose-escalation, Phase I clinical trial involving the administration of single oral doses of lodenafil carbonate. Lodenafil carbonate was administered sequentially, escalating in single doses of 1 mg - 100 mg with a washout period of at least 1 week between each dose. The progression to the next dose was allowed after clinical and laboratory exams, Ambulatory Monitoring of Arterial Pressure (AMAP) without relevant clinical modifications and adverse events without clinical relevancy. Blood samples were collected at pre-dose, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 10, 12, 14, 16, 20 and 24 h post-dosing. Plasma samples for measurement of lodenafil carbonate and lodenafil were analyzed by liquid chromatography coupled to tandem mass spectrometry. Results: No serious adverse events were observed, and none of the subjects discontinued the study due to intolerance. The AMAP measurements, clinical and laboratory exams and ECG revealed no significant changes even at higher doses. Lodenafil carbonate was not detected in any samples, indicating that it acts as a prodrug. The mean lodenafil pharmacokinetic parameters for t(max) and t(1/2) were 1.6 (+/- 0.4) h and 3.3 (+/- 1.1) h, respectively. This study demonstrated that lodenafil carbonate was well tolerated and showed a good safety profile in healthy male volunteers.

Moderna República velha: um outro ano de 1922

O objetivo deste artigo é analisar diferentes recepções da jovem República, sobretudo em seus primeiros anos. Afinal, a tradição se inscrevia em meio à modernidade, e o novo se confundia com o velho. E é junto a esse caldo de paradoxos e conflitos que se desenha a Semana de Arte Moderna - um sopro de vanguarda, de cosmopolitismo e certo otimismo, nesse contexto - mas também outras experiências sociais de caráter mais político reivindicativo. Nesse sentido, ouso expor um pouco da experiência de Lima Barreto. Não para dela fazer um exemplo que ilumina toda uma época, ou muito menos "para estragar a festa" do ano de 2012. Ao contrário, ela representa um "outro caso", outra face da mesma modernidade. Quem sabe ela sirva como testemunho do ambiente que assolou parte da intelectualidade brasileira de inícios do século, cada vez mais descrente dos destinos dessa nação, e, nesse caso, muito impactados pelos discursos raciais deterministas, os quais, após a abolição da escravidão, criavam um novo tipo de "desigualdade", dessa feita pautada na biologia.

Cem anos do direito internacional público (1913) de José mendes (1861-1918) – olhar reflexivo sobre o direito internacional nas arcadas (1911-1918)

Ensino do Direito internacional nas arcadas – José Mendes foi professor ordinário da disciplina (1911-1918) – sua obra Direito internacional público – preleções (1913) completa cem anos de publicação.

Comentários gráficos sobre os desenhos de Paulo Mendes da Rocha

Partindo-se do pressuposto de que o desenho é instrumento de diálogo entre o arquiteto e ele mesmo ou com terceiros e que este tipo de representação vem perdendo espaço para tecnologias digitais, este trabalho tem como objetivo estudar e analisar desenhos de projetos selecionados do arquiteto Paulo Mendes da Rocha, como contribuição para a discussão sobre o papel do desenho analógico no processo projetivo atual. O estudo acerca de quatro projetos (Ginásio do Clube Paulistano, Residência Butantã, Residência Millan e MuBE) se deu através de marcações realizadas pela pesquisadora sobre desenhos originais do arquiteto, com o objetivo de detectar intenções projetuais, conceitos e características dos projetos. Para tanto, foram feitas marcações gráficas sobre os desenhos originais, de maneira a evidenciar uma leitura particular da pesquisadora, permitindo assim uma melhor compreensão dos projetos escolhidos para o estudo. O desenho sobre o desenho marca os pontos que o julgamento considera importantes, tratando-se de uma leitura particular e permitindo melhor compreensão dos projetos para quem o pratica, uma vez que o ato de desenhar está estreitamente relacionado ao de pensar. Optou-se por apresentar as imagens relacionadas a citações diretas de autores

O desenho no processo projetivo: estudo das representações gráficas de projetos de Paulo Mendes da Rocha

Há anos vem-se questionando o papel da representação gráfica na arquitetura, diante das novas tecnologias gráficas computacionais e novos processos projetivos delas decorrentes. Entretanto, o desenho à mão livre ainda se mostra atuante no processo projetivo, marcado por um olhar atento, uma percepção individual e um tempo de execução que permite imersão, entrega e reflexão. Este artigo apresenta uma análise a partir de um olhar mais atento ao desenho de projetos do arquiteto Paulo Mendes da Rocha, como contribuição para a discussão sobre o papel do desenho analógico no processo projetivo. Foram selecionados alguns projetos do arquiteto: Projeto para o Concurso de remodelação do centro urbano de Santiago (Chile); Estádio Serra Dourada; Edifício Jaraguá, Caetano de Campos; Ginásio do Clube Atlético Paulistano (1958); Residência no Butantã (1964); residência Fernando Millan (1970) e Museu Brasileiro da Escultura (1986). Este artigo apresenta estudos de leitura sobre os desenhos originais do arquiteto com o objetivo de detectar as intenções projetuais, conceitos e características do projeto. Foram feitas marcações gráficas sobre os desenhos originais, evidenciando uma leitura particular do pesquisador que permitiu uma melhor compreensão dos projetos.

Inhibition of human platelet aggregation by eosinophils

AIMS: The relationship between the activity of eosinophils and platelets has been observed in recent decades by many scientists. These observations include increased numbers of eosinophils associated with platelet disorders, including changes in the coagulation cascade and platelet aggregation. Based on these observations, the interaction between eosinophils and platelets in platelet aggregation was analyze. MAIN METHODS: Human platelets were incubated with eosinophil cytosolic fraction, promyelocytic human HL-60 clone 15 cell lineage, and eosinophil cationic protein (ECP). Platelet rich plasma (PRP) aggregation was induced by adenosine diphosphate, platelet activating factor, arachidonic acid, and collagen, and washed platelets (WP) were activated by thrombin. KEY FINDINGS: Aggregation induced by all agonists was dose dependently inhibited by eosinophil cytosolic fraction. This inhibition was only partially reversed by previous incubation of the eosinophils with l-Nitro-Arginine-Methyl-Ester (l-NAME). Previous incubation with indomethacin did not prevent the cytosolic fraction induced inhibition. The separation of eosinophil cytosolic fraction by gel filtration on Sephadex G-75 showed that the inhibitory activity was concentrated in the lower molecular weight fraction. HL-60 clone 15 cells differentiated into eosinophils for 5 and 7 day were able to inhibit platelet aggregation. The ECP protein inhibited the platelet aggregation on PRP and WP. This inhibition was more evident in WP, and the citotoxicity MTT assay proved the viability of tested platelets, showing that the observed inhibition by the ECP protein does not occur simply by cell death. SIGNIFICANCE: Our results indicate that eosinophils play a fundamental role in platelet aggregation inhibition

Hydroxocobalamin quantification in human plasma by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry in a pharmacokinetic study

A rapid, sensitive and specific method for quantifying hydroxocobalamin in human plasma using paracetamol as the internal standard (IS) is described. The analyte and the IS were extracted from plasma by liquid-liquid extraction using an organic solvent (ethanol 100%; -20°C). The extracts were analyzed by high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-MS-MS). Chromatography was performed on Prevail C8 3 μm, analytical column (2.1×100 mm i.d.). The method had a chromatographic run time of 3.4 min and a linear calibration curve over the range 5-400 ng.mL-1 (r>0.9983). The limit of quantification was 5 ng.mL-1. The method was also validated without the use of the internal standard. The precision in the intra-batch validation with IS was 9.6%, 8.9%, 1.0% and 2.8% whereas without IS was 9.2%, 8.2%, 1.8% and 1.5% for 5, 15, 80 and 320 ng/mL, respectively. The accuracy in intra-batch validation with IS was 108.9%, 99.9%, 98.9% and 99.0% whereas without IS was 101.1%, 99.3%, 97.5% and 92.5% for 5, 15, 80 and 320 ng/mL, respectively. The precision in the inter-batch validation with IS was 9.4%, 6.9%, 4.6% and 5.5% whereas without IS was 10.9%, 6.4%, 5.0% and 6.2% for 5, 15, 80 and 320 ng/mL, respectively. The accuracy in inter-batch validation with IS was 101.9%, 104.1%, 103.2% and 99.7% whereas without IS was 94.4%, 101.2%, 101.6% and 96.0% for 5, 15, 80 and 320 ng/mL, respectively. This HPLC-MS-MS procedure was used to assess the pharmacokinetics of Hydroxo cobalamin following intramuscular injection 5000 μg in healthy volunteers of both sexes (10 males and 10 females). The volunteers had the following clinical characteristics (according to gender and expressed as mean ± SD [range]): males: age: 32.40 ± 8.00 y [23.00-46.00], height: 1.73 ± 0.07 m [1.62-1.85], body weight: 72.48 ± 10.22 Kg [60.20- 88.00]; females: age: 28.60 ± 9.54 y [18.00-44.00], height: 1.60 ± 0.05 m [1.54-1.70], body weight: 58.64 ± 6.09 Kg [51.70- 66.70]. The following pharmacokinetic parameters were obtained from the hydroxocobalamin plasma concentration vs. time curves: AUClast, T1/2, Tmax, Vd, Cl, Cmax and Clast. The pharmacokinetic parameters were 120 (± 25) ng/mL for Cmax, 2044 (± 641) ng.h/mL for AUClast, 8 (± 3.2) ng.mL-1 for Clast, 38 (± 15.8) hr for T1/2 and 2.5 (range 1-6) hr for Tmax. Female volunteers presented significant (p=0.0136) lower AUC (1706 ± 704) ng.h/mL) and larger (p=0.0205) clearance (2.91 ± 1.41 L/hr), as compared to male 2383 ± 343 ng.h/mL and 1.76 ± 0.23 L/hr, respectively. These pharmacokinetic differences could explain the higher prevalence of vitamin B12 deficiency in female patients. The method described validated well without the use of the internal standard and this approach should be investigated in other HPLC-MS-MS methods.