Caracterização e quantificação de marcadores químicos do extrato hidroetanólico das folhas de Kalanchoe brasiliensis Cambess

Kalanchoe brasiliensis Cambess (Crassulaceae), commonly known as saião , coirama branca , folha grossa , is originally from Brazil and commonly found in São Paulo to Bahia, mainly in the coastal zone. Regarding of biological activities, most preclinical studies were found in the literature, mainly about the anti-inflammatory activity of extracts obtained from leaves and / or aerial parts of K. brasiliensis. As regards the chemical constitution, it has been reported mainly the presence of flavonoids in the leaves of the species, but until this moment did not knows which are the active compounds. Although it is a species widely used in traditional medicine in Brazil, there is no monograph about the quality parameters of the plant drug. In this context, this study aims to characterize and quantify the chemical markers of hydroethanolic extract (HE) from the leaves of K. brasiliensis, which can be used in quality control of plant drug and derivatives obtained from this species. The methodology was divided into two parts: i. Phytochemical study: to fractionate, isolate and characterizate of the chemical (s) marker (s) of the HE from the leaves of K. brasiliensis; ii. To Developed validate of analytical method by High Performance Liquid Chromatography (HPLC)-diode array detector (DAD) to quantify the chemical (s) marker (s) of the EH. i. The EH 50% was prepared by turbo extraction method. It was then submitted to liquid-liquid partition, obtaining dichloromethane, n-butanol and ethyl acetate (AcOEt) fractions. The AcOEt fraction was selected to continue the fractionation process, because it has a chemical profile rich in flavonoids. The acOEt fraction was submitted to column chromatography using different systems for obtaining the compound Kb1. To identify this compound, it was submitted to UV analysis ii. For quantitative analysis, the EH was analyzed by HPLC, using different methods. After selecting the most appropriate method, which showed satisfactory resolution and symmetrical peaks, it was validated according to parameters in the RE 899/2003. As result, it was obtained from the AcOEt fraction the compound Kb1 (2.7 mg). Until this moment, the basic nucleus was characterized by UV analysis using shift reagents. The partial chemical structure of the compound Kb1 was identified as a flavonol, containing hydroxyls in 3 , 4 position (ring A), 5 and 7 free (ring B) and a replacement of the C3 hydroxyl by a sugar. As the analysis were performed in the HPLC coupled to a DAD, we observed that the UV spectrum of the major peaks of EH from K. brasiliensis shown similar UV spectrum. According to the literature, it has been reported the presence of patuletin glycosydes derivatives in the leaves of this species. Therefore, it is suggested that the compound Kb1 is glycosylated patuletin derivative. Probably the sugar (s) unit(s) are linked in the C3 in the C ring. . Regarding the development of HPLC analytical method, the system used consists of phase A: water: formic acid (99,7:0,3, v / v) and phase B: methanol: formic acid (99,7:0,3, v / v), elution gradient of 40% B - 58% B in 50 minutes, ccolumn (Hichrom ®) C18 (250x4, 0 mm, 5 μm), flow rate 0.8 mL / min, UV detection at 370 nm, temperature 25 ° C. In the analysis performed with the co-injection of thecompound Kb1 + HE of K. brasiliensis was observed that it is one of the major compounds with a retention time of 12.47 minutes and had a content of 15.3% in EH of leaves from K. brasiliensis. The method proved to be linear, precise, accurate and reproducible. According to these results, it was observed that compound Kb1 can be used as a chemical marker of EH from leaves of K. brasiliensis, to assist in quality control of drug plant and its derivatives

Avaliação in vitro da eficácia de produtos homeopáticos contendo momordica charantia através de bioensaios

Homeopathic medicines have been used for over two hundred years without the examination of their effects on in vivo and in vitro assays, due to the peculiarity of homeopathic preparations, the high dilution, which creates a challenge for the use of usual analytical techniques of quality control of medicine.Although there is scarcity of literature and variety of experiments, recently there have been some studies with few in vitro assays which have shown positive responses when evaluating the mechanism of action of homeopathic medicines which are able to act on a specific system.The present study aims to evaluate the efficacy of homeopathic products containing Momordica charantia through bioassays.Homeopathic products were tested by the MTT to assess cytotoxicity in RAW 264.7 (macrophage-like cells) and in tumor cells HeLa (human cervical adenocarcinoma cells), CHO K1 (Chinese hamster ovary cells), PANC-1 (human pancreas cancer cells) and PC-3 (human prostate cancer cells), dosage of inflammatory mediators NO, TNF-α and IL-6 released by RAW 264.7 cells, analysis of the death process and cell cycle changes of PC-3 by flow cytometry. The data demonstrate that homeopathic products of Momordica charantia did not show cytotoxicity to RAW 264.7, increased the production of inflammatory mediators by RAW 264.7 synergistically with LPS, showed cytotoxicity to PC-3 with change in its cell cycle inhibiting its proliferation, being the 30CH the most potent sample. Correlation studies were conducted in order to evaluate the possible in vitro applicable models to the quality control of homeopathic products with Momordica charantia. The data showed that the best applicable models in assessing the quality are the MTT to assess cytotoxicity in RAW 264.7 and PC-3 in 24 hours for Momordica charantia fruit products and dosage of NO production by RAW 264.7 with and without LPS

Desenvolvimento e validação de método analítico para determinação de WE010, WE014 e WE017

The synthetic guanylhydrazones WE010 (3,5-di-tert-butil-4-hidroxibenzaldehyde-guanylhydrazone), WE014 (4-bifenilcarboxialdehydeguanylhydrazone) and WE017 (3,4-diclorobenzaldehydeguanylhydrazone) showed high cytotoxic activity in terms of percentage inhibition of cancer cells growth. However, further progress in the development of these drug candidates requires precise and convenient methods for their qualitative and quantitative analyses. The aim of this study was to develop and validate High Performance Liquid Chromatography with diode-array detection (HPLC-DAD) and Ultra Fast Liquid Chromatography with diode-array detection (UFLC-DAD) methods suitable for as simultaneous as isolated determination of studied guanylhydrazones, based on the optimization of chromatographic parameters and obtaining reduced detection times. The chromatographic analyses of analytes by HPLC were performed on C18 ACE analytical column (150 mm x 4.6 mm), with a particle size of 5.0 μm. Among all the conditions assayed, the best results of separation were obtained with a mixture of methanol:water (60:40, v/v) as the mobile phase at a flow rate 1.5mL/min and pH of 3.5 adjusted at acetic acid. The UFLC method was developed by experimetal desing techniques in order to find optimal chromatographic analytical conditions, which were achieved on XR-ODS analytical column (50 mm x 3.0 mm), with a particle size of 2,2 μm, maintained at 25 ºC. The mobile phase was consisted of methanol:water (65:35, v/v) with 0.1% triethylamine (TEA) and pH of 3.5 adjusted at acetic acid, at a flow rate 0.5 mL/min. The procedure were validated following evaluating parameters such as specificity, linearity, limits of detection (LD) and quantification (LQ), precision, accuracy and robustness, giving results within the acceptable range. Although the UFLC method shows better sensitivity (lower values of LD and LQ), robustness (lower rates of relative standard deviation) and minimize spending time and solvent, both developed methods were adequately applied to the analysis of guanylhydrazones molecules, may be used in routine of quality control laboratories. Keywords: guanylhydrazones, HPLC/DAD, UFLC/DAD, validation of analitical method

Testes bioquímicos para avaliação do vigor em sementes de Mimosa caesalpiniaefolia Benth

Mimosa caesalpiniaefolia Benth. is a forest species of the Mimosaceae family, recommended for recovery of degraded areas. The evaluation of vigor by biochemical tests have been an important tool in the control of seed quality programs, and the electrical conductivity and potassium leaching the most efficient in the verifying the physiological potential. The objective, therefore, to adjust the methodology of the electrical conductivity test for seeds of M. caesalpiniaefolia, for then compare the efficiency of this test with the potassium in the evaluation of seed vigor of different lots of seeds M. caesalpiniaefolia. To test the adequacy of the electrical conductivity were used different combinations of temperatures , 25 °C and 30 ºC, number of seeds , 25 and 50, periods of imbibition , 4 , 8 , 12 , 16 and 24 hours , and volumes deionized water, 50 mL and 75mL. For potassium leaching test, which was conducted from the results achieved by the methodology of the adequacy of the electrical conductivity test, to compare the efficiency of both tests , in the classification of seeds at different levels of vigor, and the period 4 hours also evaluated because the potassium leaching test can be more efficient in the shortest time . The best combination obtained in experiment of electrical conductivity is 25 seeds soaked in 50 mL deionized or distilled water for 8 hours at a temperature of 30 ° C. Data were subjected to analysis of variance, the means were compared with each other by F tests and Tukey at 5 % probability, and when necessary polynomial regression analysis was performed. The electrical conductivity test performed at period eight hour proved to be more efficient in the separation of seed lots M. caesalpiniaefolia at different levels of vigor compared to the potassium test

Obtenção de insumo farmacêutico a partir das folhas de Cissampelos sympodialis Eichl e seu efeito sobre mediadores inflamatórios relevantes para a asma

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Testes de vigor para avaliação de sementes de pepino

Dentro de um programa de controle de qualidade, a avaliação do vigor de sementes é fundamental e necessária para o sucesso da produção. O trabalho teve por objetivo comparar diferentes métodos para avaliação do potencial fisiológico de sementes de pepino. Para tanto, cinco lotes de sementes do híbrido Safira foram submetidos aos seguintes testes: germinação, primeira contagem de germinação, condutividade elétrica (4x50 sementes; 25ºC; 50 e 75mL; 12, 18 e 24h), germinação a baixa temperatura (4x50 sementes; 18ºC; contagens aos quatro e oito dias após a semeadura); envelhecimento acelerado tradicional (41ºC; 48, 72 e 96h) e com solução saturada de NaCl (41ºC; 72 e 96h). Pelos resultados obtidos, pode-se concluir que os testes de condutividade elétrica (50 e 75mL; 12, 18 e 24h) e de envelhecimento acelerado com o uso de solução salina (41ºC, por 72h), possibilitam a melhor identificação de lotes com diferentes níveis de qualidade fisiológica.

Temperatura e período de exposição de sementes de milho no teste de envelhecimento acelerado

O teste de envelhecimento acelerado (EA) apresenta possibilidades de uso nos programas de controle de qualidade de sementes de milho. Porém, para uso em rotina, há necessidade de padronização das condições de temperatura e período de exposição das sementes na realização do teste. Esta pesquisa teve por objetivo avaliar o efeito da combinação de duas temperaturas (42 e 45ºC) e dois períodos de envelhecimento (72 e 96h) nos resultados do teste de envelhecimento acelerado em sementes de milho. Para isso, sementes de seis genótipos de milho, tratadas e sem tratamento fungicida, com altos valores de germinação inicial, semelhantes entre si, dentro de cada genótipo, foram submetidas ao EA, de acordo com condições de temperatura e período de exposição pré-estabelecidos. Utilizou-se o delineamento experimental inteiramente casualizado com os tratamentos (lotes x germinação, antes e após o EA) dispostos em esquema fatorial, empregando-se o teste de Tukey a 5% de probabilidade na comparação das médias. Verificou-se que sementes de milho apresentam diferenças de sensibilidade ao envelhecimento acelerado, condicionadas pelo genótipo e tratamento fungicida. Contudo, a combinação 45ºC por 72h possibilita separar lotes de qualidade fisiológica superior e inferior, independentemente do genótipo. O período de exposição das sementes ao envelhecimento acelerado determina o teor de água atingido pelas mesmas.

Metodologia alternativa para condução do teste de envelhecimento acelerado em sementes de milho

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Envelhecimento acelerado como teste de vigor para sementes de milho e soja

A avaliação do vigor de sementes tem sido ferramenta fundamental dentro do programa de controle de qualidade, sendo o teste de envelhecimento acelerado (EA) parte importante do processo. O trabalho foi desenvolvido com o objetivo de confirmar metodologia específica para avaliação do vigor de sementes, em laboratório, tendo como base o EA. Utilizou sementes de milho e soja, seis lotes para cada espécie, com padrões de qualidade (germinação) comercialmente aceitas. As condições de envelhecimento préestabelecidas foram 42ºC/96h e 45ºC/72h para milho e 41ºC/48 e 72h e 42ºC/48 e 72h para soja. Para cada combinação de temperatura e período de envelhecimento, as sementes foram colocadas em caixas plásticas, sobre tela, com 40mL de água destilada, distribuídas em camada única e com base no peso das mesmas. O envelhecimento foi feito usando-se dois tipos de câmara, BOD e jaquetada de água (CJ). Foi determinado o teor de água (TA) e a germinação das sementes antes e após o EA. A absorção de água pelas sementes considerando-se todas as combinações de período de exposição e temperatura, as duas câmaras e as duas formas de determinação do TA, foi maior quando as sementes foram acondicionadas em camada única em relação aquelas pesadas (massa constante), usando-se a câmara jaquetada. Para o milho, a combinação de 45ºC/72h com o acondicionamento das sementes em camada única foi o método mais eficiente para a separação dos lotes quanto ao vigor. Para soja, a combinação 42ºC/48h foi mais eficiente na separação dos lotes, não havendo interferência do método de acondicionamento das sementes, na tela. Conclui-se que a camada única e a câmara tipo BOD devem ser preferencialmente utilizadas no teste EA face aos menores custos de aquisição e manutenção da BOD, visando à separação de lotes quanto aos níveis de vigor, para milho e soja.

Avaliação do potencial fisiológico de lotes de sementes de soja

Vários procedimentos são utilizados para avaliar o potencial fisiológico de sementes, sendo fundamental a utilização daqueles que reflitam melhor o desempenho em campo. A pesquisa teve como objetivo avaliar o potencial fisiológico de sementes de soja por meio de testes de avaliação rápida, de vigor, de emergência de plântulas em campo e análise computadorizada de imagens (SVIS®), procurando identificar o procedimento mais eficiente na separação de lotes. Cinco lotes das cultivares BRS Valiosa RR e CD 208 foram avaliados em duas épocas (antes e após armazenamento) pelos testes de germinação, de avaliação rápida (embebição em água e pH do exsudato), de vigor (envelhecimento acelerado e condutividade elétrica), de emergência de plântulas em campo e análise computadorizada de imagens (SVIS®). Os testes de germinação em areia, de condutividade elétrica e de embebição em água foram eficientes na separação dos lotes, mas apenas este último possibilitou identificar diferenças consistentes entre os lotes nos dois períodos avaliados. Assim, o teste de embebição em água pode ser considerado promissor na composição de programas de controle de qualidade.

Alternative procedure for the cold test for soybean seeds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Automated system of seedling image analysis (SVIS) and electrical conductivity to assess sun hemp seed vigor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Adaptação de um sistema automatizado para medição de microdureza

The hardness test is thoroughly used in research and evaluation of materials for quality control. However, this test results are subject to uncertainties caused by the process operator in the moment of the mensuration impression diagonals make by the indenter in the sample. With this mind, an automated equipment of hardness mensuration was developed. The hardness value was obtained starting from the mensuration of plastic deformation suffered by the material to a well-known load. The material deformation was calculated through the mensuration of the difference between the progress and retreat of a diamond indenter on the used sample. It was not necessary, therefore, the manual mensuration of the diagonals, decreasing the mistake source caused by the operator. Tension graphs of versus deformation could be analyzed from data obtained by the accomplished analysis, as well as you became possible a complete observation of the whole process. Following, the hardness results calculated by the experimental apparatus were compared with the results calculated by a commercial microhardness machine with the intention of testing its efficiency. All things considered, it became possible the materials hardness mensuration through an automated method, which minimized the mistakes caused by the operator and increased the analysis reliability

Procedimento para condução do teste de envelhecimento acelerado em sementes de algodão

The evaluation of the effect of seed has been fundamental tool in the program of quality control, and the accelerated aging test, an important part of that process. Some authors report that the germination and vigor are associated with seed size and facing the lack of consensus for the use of this test methodology and the relationship of seed size on the physiological quality, this work aimed to adapt the existing methodology as the temperature and period of conditioning on the performance of two sizes of seed cotton. The seeds were classified into two meshes of sieves, and then subjected to tests of water content, germination and electrical conductivity. The experimental design was completely randomized, analyzed statistically in a factorial threefold. Seeds of different sizes, showed no difference in percentage of germination. Seeds subjected to 40 degrees C for 42h of conditioning showed better percentage of germination.

A engenharia simultânea e o perfil da indústria de confecção da grande Natal/RN utilizando as ferramentas CAD/PDM e ERP

This work presents the research carried through in the industrial segment of confection of clothes of the Great Natal whose objective is to show the profile, enterprise and technological management as also the use of simultaneous engineering in the development of products. The research approaches two studies. The first one presents the current picture of the companies, synthesized through twelve variable. As, through fifteen variable it shows to the level of use of Simultaneous Engineering in the Development of Products and its amplitude in relation to the Integrated Management using tools CAD, PDM and ERP (Computer Aided Design, Product Management Date, Enterprise Resource Planning). The integration of these systems acts aiming the reduction of the cost and the development time of products. The reached results indicate that simultaneous engineering is a competitive advantage and becomes possible: to reduce the life cycle of the product, to rationalize the resources, to incorporate one high standard of the quality to the process and product as well as to personalize the product to take care of the global market. It is important to note that this work also is considered to contribute for the better understanding of the real companies situation of confection located at the Great Natal and its role in the economy of the State of the Rio Grande do Norte