Phenolic Acids and Flavonoids from Pithecellobium dulce (Robx.) Benth Leaves Exhibit Ovicidal Activity against Haemonchus contortus.

Pithecellobium dulce (Robx.) Benth is an arboreal legume used in traditional medicine for the treatment of several ailments, including a number of intestinal disorders, and as a natural deworming. The objective of this study was to evaluate the ovicidal activity of a hydroalcoholic extract (HA-E) and its fractions (aqueous, Aq-F and organic, EtOAc-F) from P. dulce leaves, as well as subfractions (C1F1-C17) obtained from EtOAc-F against Haemonchus contortus eggs. The HA-E, Aq-F, and EtOAc-F were assessed at 0.03-5.00 mg/mL and subfractions (C1F1-C17) were assessed at 0.06-1.00 mg/mL. The HA-E and Aq-F showed an ovicidal activity close to 100% at 2.5 mg/mL, and EtOAc-F displayed the highest anthelmintic effect (100% at 0.25 mg/mL). Meanwhile, the sub-fractions with the highest ovicidal effect were C1F6, C1F9, C1F10, C1F11, and CIF13. The main compounds identified in the most active fractions (C1F9, C1F11, and C1F13) were kaempferol (1), quercetin (2), coumaric acid (3), ferulic acid (4), luteolin 7-O-rhamnoside (5), quercetin 3-O-rhamnoside (6), and a caffeoyl derivate (NI). The results indicate that P. dulce leaves exhibit a potent anthelmintic activity and contain bioactive compounds able to inhibit egg hatching in H. contortus. Therefore, this plant could be used for the control of gastrointestinal nematodes in small ruminants.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark / Caracterização fitoquímica, atividade antioxidante e antibacteriana do extrato hidroetanólico de cascas do caule de Anadenanthera peregrina

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-¹. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.(AU)

O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência(HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.(AU)

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark / Caracterização fitoquímica, atividade antioxidante e antibacteriana do extrato hidroetanólico de cascas do caule de Anadenanthera peregrina

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-¹. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência(HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark

Abstract The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

Resumo O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico-química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark / Caracterização fitoquímica, atividade antioxidante e antibacteriana do extrato hidroetanólico de cascas do caule de Anadenanthera peregrina

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físicoquímica, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Validation of an analytical method for the quantification of human fibrinogen in pharmaceutical products by size-exclusion liquid chromatography (SEC-HPLC).

Fibrinogen plays a vital role in normal homeostasis by promoting platelet aggregation, clot formation and fibrinolysis. It is quantified in finished pharmaceutical products using different methods described in pharmacopoeia, but these are inaccurate, difficult to validate and do not allow for identification of aggregates or protein products of the same formulation. The aim of this study was to develop and validate a method for quantification of the content of fibrinogen and other proteins present in pharmaceutical formulations by comparing it with current pharmacopeial methods. Fibrinogen was quantified in two commercial products and compared to a pharmacopeial method using a validated method for size-exclusion high-pressure liquid chromatography (SEC-HPLC). The fibrinogen level was in accordance with both products' specifications. The SEC-HPLC method showed that the percentage of fibrinogen was 94.88 for one product and 50.68 for the other, and detected high molecular weight aggregates in the second product. The SEC-HPLC method that we developed is an improvement to the current pharmacopeial method, because it allows for quantification of fibrinogen and determination of product purity. This is important because greater purity can reduce potential adverse effects of pharmaceutical products in patients.

Analysis of Malassezia Lipidome Disclosed Differences Among the Species and Reveals Presence of Unusual Yeast Lipids.

Malassezia yeasts are lipid dependent and part of the human and animal skin microbiome. However, they are also associated with a variety of dermatological conditions and even cause systemic infections. How these yeasts can live as commensals on the skin and switch to a pathogenic stage has long been a matter of debate. Lipids are important cellular molecules, and understanding the lipid metabolism and composition of Malassezia species is crucial to comprehending their biology and host-microbe interaction. Here, we investigated the lipid composition of Malassezia strains grown to the stationary phase in a complex Dixon medium broth. In this study, we perform a lipidomic analysis of a subset of species; in addition, we conducted a gene prediction analysis for the detection of lipid metabolic proteins. We identified 18 lipid classes and 428 lipidic compounds. The most commonly found lipids were triglycerides (TAG), sterol (CH), diglycerides (DG), fatty acids (FAs), phosphatidylcholine (PC), phosphatidylethanolamine (PE), ceramides, cholesteryl ester (CE), sphingomyelin (SM), acylcarnitine, and lysophospholipids. Particularly, we found a low content of CEs in Malassezia furfur, atypical M. furfur, and Malassezia pachydermatis and undetectable traces of these components in Malassezia globosa, Malassezia restricta, and Malassezia sympodialis. Remarkably, uncommon lipids in yeast, like diacylglyceryltrimethylhomoserine and FA esters of hydroxyl FAs, were found in a variable concentration in these Malassezia species. The latter are bioactive lipids recently reported to have antidiabetic and anti-inflammatory properties. The results obtained can be used to discriminate different Malassezia species and offer a new overview of the lipid composition of these yeasts. We could confirm the presence and the absence of certain lipid-biosynthesis genes in specific species. Further analyses are necessary to continue disclosing the complex lipidome of Malassezia species and the impact of the lipid metabolism in connection with the host interaction.

Simultaneous determination of anti-diabetic drugs

A novel reverse phase, isocratic HPLC method is described to separate five anti-diabetic drugs i.e., glimepiride, metformin, sitagliptin, rosiglitazone and pioglitazone. Nucleosil C18 analytical column was used as stationary phase, while mobile phase consisted of acetonitrile:phosphate buffer: methanol (40/40/20, v/v) pH 2.0. Effluent was monitored at a flow rate 1 mL/min and detected at wavelength of 240 nm. This research produced excellent chromatography over a wide concentration range of 25-10000 ng/mL. Sepprated and well resolved quantifiable peaks were obtained and test results were linear in this range. Correlation coefficient of more than 0.9990 was witnessed as well as Low %RSD values i.e., maximum 2.0% documented excellent precision of the method. Good recoveries from pharmaecutical (99-101%), urine and plasma samples (>96%) in a range of concentrtion granted very good linearity, accuracy and precision. The projected method has satisfactory applications in quality control of these molecules as well as quantification of these molecules in urine and plasma samples.

Análise de limite de galactosamina em hexosaminas totais em amostras injetáveis de heparina sódica suína / Galactosamine limit analysis in total hexosamines in injectable porcine sodium heparin samples

Introdução: A pesquisa, uma das vertentes da vigilância sanitária, tem sua importância justificada pela busca de respostas aos diversos prejuízos relativos à saúde. A heparina, produto biológico com propriedades anticoagulantes e antitrombóticas, esteve relacionada com eventos adversos entre 2007 e 2008. Diante do ocorrido, os compêndios oficiais atualizaram a monografia para matéria-prima. No entanto, há uma deficiência de monografias para avaliação do produto final. Objetivo: Propor método físico-químico de análise de limite de galactosamina em hexosaminas totais a partir do produto acabado de heparina sódica suína. Método: Foi desenvolvido método analítico a partir da cromatografia líquida de alta eficiência por troca iônica e detecção amperométrica, com as devidas avaliações de adequação do sistema para posterior análise de três amostras de heparina sódica suína, as quais foram previamente submetidas ao protocolo de preparo de amostra através da coluna Micro Bio-Spin™. Resultados: As amostras foram comparadas com a solução de adequação do sistema e matéria-prima de heparina sódica suína, sendo possível detectar presença de galactosamina em uma das três amostras analisadas em quantidade inferior ao limite estipulado pela Farmacopeia Americana. Conclusões: Conclui-se que o método é eficiente para análise do produto acabado e, por isso, será sugerido à Farmacopeia Brasileira.

Introduction: Research, one of the core areas of health surveillance, has its importance justified by its search for answers to various health problems. Heparin, a biological product with anticoagulant and antithrombotic properties, has been related to adverse events between the years 2007­2008. Because of that, the official compendiums updated the monograph for raw material. However, there is a lack of monographs that evaluate the final product. Objective: The goal of this study is to propose a physicochemical method of analysis of the limits of galactosamine in total hexosamine from the finished product of porcine sodium heparin. Method: We developed an analytical method from the ionexchange high performance liquid chromatography with amperometric detection, with the appropriate evaluations of the system suitability for posterior analysis of three samples of porcine sodium heparin previously submitted to the sample preparation protocol through the Micro Bio-Spin column. Results: The samples were compared to the solution of the system suitability and raw material of porcine sodium heparin. We could detect the presence of galactosamine in one of the three analyzed samples in lower amounts than the limit stipulated by the American pharmacopeia. Conclusions: We concluded that the aforementioned method is efficient for the analysis of the finished product and that is the reason why it will be suggested to the Brazilian pharmacopeia.

A comparative study of two separate analytical techniques for the simultaneous determination of diclofenac sodium and diacerein from combined dosage form

ABSTRACT Diclofenac sodium (DS) and diacerein (DC) have emerged as a potential combination therapy for the treatment of knee osteoarthritis. Therefore a validated analytical method is essential for the simultaneous estimation of both from combined dosage form. A ratio derivative spectrophotometric and a chromatographic technique have been developed for the simultaneous determination of DS and DC. The quantification was done at 263.00 nm for DC and 304.50 nm for DS in the first method, whereas 257 nm for DC and at 274 nm for DS for LC-DAD analysis in chromatographic method using acetate buffer and methanol as the mobile phase at a flow-rate 0.50 mL/min. Both of these methods are found to be linear in the concentration range under study with r2 value 0.999 and 0.996 for DS and DC respectively in ratio derivative spectroscopy and 0.998 and 0.999 for DS and DC respectively in LC-DAD study. Both of these methods are found to be accurate and precise, though greater robustness and precision is observed with chromatographic analysis over the ratio derivative spectroscopy. Statistically there was no significant difference between proposed ratio derivative spectrophotometric and LC-DAD methods.

Physical-chemical and biological characterization of different preparations of equine chorionic gonadotropin.

Ovarian stimulation with commercial preparations of equine chorionic gonadotropin (eCG) produces extremely variable responses in domestic animals, ranging from excessive stimulation to practically no stimulation, when applied on the basis of their declared unitage. This study was conducted to analyze four commercial preparations from different manufacturers via reversed-phase HPLC (RP-HPLC) in comparison with a reference preparation and an official International Standard from the World Health Organization. The peaks obtained by this qualitative and quantitative physical-chemical analysis were compared using an in vivo bioassay based on the ovarian weight gain of prepubertal female rats. The RP-HPLC data showed one or two peaks close to a main peak (tR= 27.9 min), which were related to the in vivo bioactivity. Commercial preparations that have this altered peak showed very little or no in vivo activity, as demonstrated by rat ovarian weight and in peripubertal gilts induced to ovulate. Overall, these findings indicate that RP-HPLC can be a rapid and reliable tool to reveal changes in the physicochemical profile of commercial eCG that is apparently related to decreased biological activity of this hormone.

Rapid and sensitive ultra-high-pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study.

Benznidazole (BNZ) and nifurtimox are the only drugs available for treating Chagas disease. In this work, we validated a bioanalytical method for the quantification of BNZ in plasma aimed at improving sensitivity and time of analysis compared with the assays already published. Furthermore, we demonstrated the application of the method in a preclinical pharmacokinetic study after administration of a single oral dose of BNZ in Wistar rats. A Waters® Acquity UHPLC system equipped with a UV-vis detector was employed. The method was established using an Acquity® UHPLC HSS SB C18 protected by an Acquity® UHPLC HSS SB C18 VanGuard guard column and detection at 324 nm. The mobile phase consisted of ultrapure water-acetonitrile (65:35), and elution was isocratic. The mobile phase flow rate was 0.55 mL/min, the volume of injection was 1 µL, and the run time was just 2 min. The samples were kept at 25°C until injection and the column at 45°C for the chromatographic separation. The sample preparation was performed by a rapid protein precipitation with acetonitrile. The linear concentration range was 0.15-20 µg/mL. The pharmacokinetic parameters of BNZ in rats were determined and the method was considered sensitive, fast and suitable for application in pharmacokinetic studies.

The antidepressant-like effect of ethynyl estradiol is mediated by both serotonergic and noradrenergic systems in the forced swimming test.

17α-Ethynyl-estradiol (EE2, a synthetic steroidal estrogen) induces antidepressant-like effects in the forced swimming test (FST) similar to those induced by 5-HT and noradrenaline reuptake inhibitors (dual antidepressants). However, the precise mechanism of action of EE2 has not been studied. In the present study, the participation of estrogen receptors (ERs) and the serotonergic and the noradrenergic presynaptic sites in the antidepressant-like action of EE2 was evaluated in the FST. The effects of the ER antagonist ICI 182,780 (10 µg/rat; i.c.v.), the serotonergic and noradrenergic terminal destruction with 5,7-dihydroxytryptamine (5,7-DHT; 200 µg/rat, i.c.v.), and N-(2-chloro-ethyl)-N-ethyl-2-bromobenzylamine (DSP4; 10mg/kg, i.p.) were studied in ovariectomized rats treated with EE2 and subjected to the FST. In addition, the participation of α2-adrenergic receptors in the antidepressant-like action of EE2 was explored using the selective α2-receptor antagonist idazoxan (0.25, 0.5 and 1.0mg/kg, i.p.). EE2 induced an antidepressant-like action characterized by a decrease in immobility behavior with a concomitant increase in swimming and climbing behaviors. The ER antagonist, 5,7-DHT, DSP4, and idazoxan blocked the effects of EE2 on the immobility behavior, whereas ICI 182,780 and 5,7-DHT affected swimming behavior. The noradrenergic compound DSP4 altered climbing behavior, while Idazoxan inhibited the increase of swimming and climbing behaviors induced by EE2. Our results suggest that the antidepressant-like action of EE2 implies a complex mechanism of action on monoaminergic systems and estrogen receptors.

Poly (Methyl Methacrylate) absorption and releasing of nystatin and fluconazole / Absorção e liberação de nistatina e fluconazol por polimetilmetacrilato

Objectives: The aim of this study was to verify if there are poly (methyl methacrylate) (PMMA) absorptionand releasing of nystatin (NYS) and fluconazole (FLZ) after simulated treatment of oral candidosis. Materialsand methods: Specimens (30 × 25 × 5 mm) prepared with PMMA polymerized by hot water bath or microwaveenergy were immersed into NYS (3.12 μg/mL), FLZ (2.56 μg/mL) or deionized water (control) during14 days at 35 ± 2 °C. After treatment simulation, specimens were immersed into distilled water during 3, 7, 10and 14 days. The immersion liquid was changed after each analysis. Higher performance liquid chromatographywas used in order to detect antifungal compounds. In order to determine if there was surface depositionof drugs on PMMA resin, specimens were analyzed with electronic microscopy (SEM). Results: None of theantifungal agents was released from the PMMA resins. Conclusion: Within the limitations of this study, itcould be concluded that PMMA resins had no drug absorption with posterior release.

Objetivos: O objetivo deste estudo foi verificar se o poli (metil metacrilato) (PMMA) é capaz de absorver e liberar nistatina (NYS) e fluconazol (FLZ) após simular um tratamento para candidose oral. Materiais e métodos:Espécimes (30 × 25 × 5 mm) foram preparados em resina de PMMA por banho de água quente ou energia demicro-ondas e, em seguida, imersos em solução contendo NYS (3.12 μg/mL), FLZ (2.56 μg/mL) ou água deionizada(controle) durante 14 dias a 35 ± 2 °C. Após a simulação de tratamento, os espécimes foram imersos em água destilada durante 3, 7, 10 e 14 dias. O líquido de imersão foi trocado após cada análise. Cromatografia líquida de alta performance foi utilizada para detectar a presença dos agentes antifúngicos. Para determinar se houve deposição dos agentes antifúngicos na superfície de PMMA, os espécimes foram analisados por microscopia eletrônica de varredura(MEV). Resultados: Não houve liberação de agentes antifúngicos dos espécimes. Conclusão: Considerando as limitações deste estudo, pode-se concluir que a resina de PMMA não absorve ou libera agentes antifúngicos.

Development studies of captopril certified reference material / Estudos de desenvolvimento de captopril material de referência certificado

This paper describes the studies performed with the candidate Certified Reference Material (CRM) of captopril, the first CRM of an active pharmaceutical ingredient (API) in Brazil, including determination of impurities (organic, inorganic and volatiles), homogeneity testing, short- and long-term stability studies, calculation of captopril content using the mass balance approach, and estimation of the associated measurement uncertainty.

Este artigo descreve os estudos realizados com o candidato a Material de Referência Certificado (MRC) de captopril, primeiro MRC de fármacos no Brasil, incluindo a determinação de impurezas (orgânicas, inorgânicas e voláteis), testes de homogeneidade, testes de estabilidade de curta e longa duração, cálculo do teor de captopril por balanço de massa e estimativa da incerteza de medição associada ao valor certificado.