Estudo fitoquímico, morfoanatômico e avaliação de atividades biológicas e propriedades antioxidantes de Ocotea diospyrifolia (Meisn.) Mez

Orientador : Profª Drª Josiane de F.G. DiasCoorientadores : Prof. Dr. Obdulio G. Miguel, Profª Drª Sandra M.W. ZaninDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Síde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa : Curitiba, 26/02/2018Inclui referências : p. 100-118Área de concentração : Insumos, medicamentos e correlatosResumo : O Brasil abriga inúmeras espécies de plantas, as quais representam uma promissora fonte de descoberta de moléculas bioativas. A espécie Ocotea diospyrifolia (Meisn.) Mez. pertence ao gênero Ocotea Aubl. e a família Lauraceae, grupos que tem demonstrado grande potencial farmacológico. Assim, neste trabalho objetivou-se estudar a composição fitoquímica da espécie citada, bem como avaliar suas características morfoanatômicas e atividades antioxidante e biológicas. Para este fim, folhas e galhos foram coletados no Capão do Cifloma (Curitiba-PR), a partir dos quais foram feitas as análises morfoanatômicas e de teor de cinzas e umidade, além dos extratos e fracionamentos em aparelho de Soxhlet. Ainda, a partir das folhas, foi realizada a extração do óleo essencial por hidrodestilação, e sua posterior caracterização por CG-EM. A partir dos extratos brutos e frações hexano foi realizada a quantificação de clorofilas totais e carotenoides. Essas amostras, juntamente com as demais frações e óleo essencial, foram analisadas quanto ao seu potencial antioxidante pelas metodologias de formação do complexo fosfomolibdênio, redução do radical 2,2-difenil-1-pictilhidrazila (DPPH) e peroxidação lipídica. Além disso, as amostras foram avaliadas quanto a sua toxicidade frente a Artemia salina, capacidade hemolítica e fitotoxicidade. Também foram aplicadas técnicas de separação, no intuito de isolar e/ou identificar compostos na espécie. Os resultados obtidos evidenciaram características morfoanatômicas condizentes com o descrito anteriormente para a espécie. O óleo essencial foi extraído a partir de duas coletas em períodos distintos, revelando a presença do espatulenol entre os componentes majoritários das duas amostras. Os ensaios antioxidantes demonstraram uma melhor atividade para as frações acetato de etila e etanólica dos galhos, e uma potencial atividade tóxica para o óleo essencial e a fração hexano do caule. As técnicas de separação aplicadas levaram ao isolamento do aminoácido 4-hidróxi-N-metilprolina da fração remanescente dos galhos. A partir disso, sugere-se que o estudo da O. diospyrifolia seja continuado, visto a possibilidade de aplicabilidade de seus extratos, frações óleo essencial e possíveis compostos isolados na área da farmacologia. Palavras-chave: Lauraceae. Óleo essencial. Espatulenol. 4-hidróxi-N-metilprolina. Toxicidade.Abstract : Brazil is home to innumerous plant species, which represent a promising source for finding new bioactive molecules. The species Ocotea diospyrifolia (Meisn.) Mez. belongs to the Ocotea Aubl. genus and Lauraceae family, groups that have been demonstrating huge pharmacological potential. Therefore, this work aimed to evaluate the phytochemical composition of the named species, as well as its morphoanatomical characteristics, antioxidant and biological activities. To this end, leaves and stem barks were collected in the Capão do Cifloma (Curitiba-PR), from where it was made morphoanatomical, ash and umidity content analysis, as well as the extracts and fractions in a Soxhlet apparatus. In addition, from the leaves, it was extracted the essential oils by hydrodistillation, and its further characterization by CGMS. From the crude extracts and hexane fractions it was performed the quantification of total chlorophylls and carotenoids. These samples, together with the remaining fractions and essential oil were analysed for their antioxidant potential through the molibdenium complex formation, radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) reduction and lipid peroxidation methods. Moreover, samples were evaluated for their toxicity against Artemia salina, hemolytic capacity and phytotoxicity. Separation techniques were also applied, in order to isolate and/or identify compounds in the species. The results obtained highlighted morphoanatomical characteristics similar to the ones described previously for the species. The essential oil was extracted from two collections in distinct periods, revealing the presence of spathulenol as one of the major constituents in both samples. The antioxidant assays demonstrated a better activity for ethyl acetate and etanolic fraction from the stem bark, and a potential toxic activity for the essential oil and stem bark hexane fraction. Separation techniques applied lead to isolation the aminoacid 4-hydroxy-N-methylproline, from the stem bark remanescent fraction. From this, it is suggested that the study of O. disospyrifolia should be continued, since there is a possibility of applicability from its extracts, fractions, essential oil and possible isolated compounds in the pharmacology area. Keywords: Lauraceae. Essential oil. Spathulenol. 4-hydroxy-N-methylproline. Toxicity

Contribution to the identification of the species Myrcia hatschbachii D. Legrand (Myrtaceae): anatomical and histochemical analyses

The genus Myrcia is used in folk medicine to treat diabetes. The plants used in folk medicine require morphological and anatomical references to attest to its authenticity. This is the first report of the microscopic study of Myrcia hatschbachii. In this sense, the aim of the present study was to describe the anatomical characteristics, in order to contribute to the species' identification. For anatomical and surface analyses free-hand sections were prepared and observed using optical microscopy, simultaneously some materials were processed and observed under scanning electron microscopy. In addition, histochemical tests were performed. The anatomical features described here correspond with previously reported features found in other species of the genus Myrcia. Furthermore, the following anatomical markers were observed in Myrcia hatschbachii: druses in the spongy parenchyma, concave-convex midrib, heart-shaped petiole, oval-shaped stem, C-shaped vascular bundle in the midrib and open arc shape with invaginated ends in the petiole; phenolic compounds in the phloem and lignified cells in the pith. The results of anatomical and histochemical analyses provide micromorphological and microchemical features that can help in the taxonomy and proper identification of the species

Chemical composition, phytotoxic potential, biological activities and antioxidant properties of Myrcia hatschbachii D. Legrand essential oil

Myrcia hatschbachii D. Legrand (Myrtaceae) is a native and endemic species from Brazil. This study investigated the essential oil composition, phytotoxic potential, in vitro toxicity, antioxidant properties and antibacterial activity of species. Chromatographic analysis of essential oil identified trans-calamanene, (E)-caryophyllene and spathulenol as major components. Antioxidant capacity was determined by the DPPH• scavenging method and phosphomolybdenum complex formation assay. Antibacterial activity was evaluated using the Minimal Inhibitory Concentration test, demonstrating that the essential oil was active toward Enterococcus faecalis and Staphylococcus aureus. Phytotoxic activity of essential oil was analyzed by testing interference on germination and growth of Lactuca sativa, demonstrating significant inhibition of the hypocotyls and radicles of seeds. Preliminary toxicity studies were determined using Artemia salina, resulting in an LC50 of 409.92 µg/mL, and through hemolytic activity. The results of the phytotoxic activity point to a possible application for Myrcia hatschbachii in the development of natural herbicides and the in vitro toxicity assays suggests the performance of antitumoral activity tests, having in mind the prospection of antineoplastic drugs

Composition, antioxidant properties, and biological activities of the essential oil extracted from Ocotea diospyrifolia (Meisn.) Mez.

The article has aimed to characterize the essential oil extracted from Ocotea diospyrifolia (Meisn.) Mez. leaves, in terms of its chemical composition and antioxidant, hemolytic, and phytotoxic potentials, as well as its toxicity against Artemia salina. The major constituents identified by CG-MS were δ-elemene, spathulenol, and β-atlantol. When screened for potential biological activities, the essential oil presented low toxicity against Artemia salina, and a capacity of lysing red blood cells. Also, the evaluation of its in vitro antioxidant activity, using the phosphomolybdenum method, showed better results when compared to butylhydroxytoluene (BHT) and rutin. In conclusion, the results obtained showed certain in vitro toxicity, leading to an interesting target for cytotoxicity evaluations of carcinoma cells

Estudo fitoquímico, morfoanatômico e avaliação de atividades biológicas e propriedades antioxidantes de Ocotea diospyrifolia (Meisn.) Mez

Orientador : Profª Drª Josiane de F.G. DiasCoorientadores : Prof. Dr. Obdulio G. Miguel, Profª Drª Sandra M.W. ZaninDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências da Síde, Programa de Pós-Graduação em Ciências Farmacêuticas. Defesa : Curitiba, 26/02/2018Inclui referências : p. 100-118Área de concentração : Insumos, medicamentos e correlatosResumo : O Brasil abriga inúmeras espécies de plantas, as quais representam uma promissora fonte de descoberta de moléculas bioativas. A espécie Ocotea diospyrifolia (Meisn.) Mez. pertence ao gênero Ocotea Aubl. e a família Lauraceae, grupos que tem demonstrado grande potencial farmacológico. Assim, neste trabalho objetivou-se estudar a composição fitoquímica da espécie citada, bem como avaliar suas características morfoanatômicas e atividades antioxidante e biológicas. Para este fim, folhas e galhos foram coletados no Capão do Cifloma (Curitiba-PR), a partir dos quais foram feitas as análises morfoanatômicas e de teor de cinzas e umidade, além dos extratos e fracionamentos em aparelho de Soxhlet. Ainda, a partir das folhas, foi realizada a extração do óleo essencial por hidrodestilação, e sua posterior caracterização por CG-EM. A partir dos extratos brutos e frações hexano foi realizada a quantificação de clorofilas totais e carotenoides. Essas amostras, juntamente com as demais frações e óleo essencial, foram analisadas quanto ao seu potencial antioxidante pelas metodologias de formação do complexo fosfomolibdênio, redução do radical 2,2-difenil-1-pictilhidrazila (DPPH) e peroxidação lipídica. Além disso, as amostras foram avaliadas quanto a sua toxicidade frente a Artemia salina, capacidade hemolítica e fitotoxicidade. Também foram aplicadas técnicas de separação, no intuito de isolar e/ou identificar compostos na espécie. Os resultados obtidos evidenciaram características morfoanatômicas condizentes com o descrito anteriormente para a espécie. O óleo essencial foi extraído a partir de duas coletas em períodos distintos, revelando a presença do espatulenol entre os componentes majoritários das duas amostras. Os ensaios antioxidantes demonstraram uma melhor atividade para as frações acetato de etila e etanólica dos galhos, e uma potencial atividade tóxica para o óleo essencial e a fração hexano do caule. As técnicas de separação aplicadas levaram ao isolamento do aminoácido 4-hidróxi-N-metilprolina da fração remanescente dos galhos. A partir disso, sugere-se que o estudo da O. diospyrifolia seja continuado, visto a possibilidade de aplicabilidade de seus extratos, frações óleo essencial e possíveis compostos isolados na área da farmacologia. Palavras-chave: Lauraceae. Óleo essencial. Espatulenol. 4-hidróxi-N-metilprolina. Toxicidade.Abstract : Brazil is home to innumerous plant species, which represent a promising source for finding new bioactive molecules. The species Ocotea diospyrifolia (Meisn.) Mez. belongs to the Ocotea Aubl. genus and Lauraceae family, groups that have been demonstrating huge pharmacological potential. Therefore, this work aimed to evaluate the phytochemical composition of the named species, as well as its morphoanatomical characteristics, antioxidant and biological activities. To this end, leaves and stem barks were collected in the Capão do Cifloma (Curitiba-PR), from where it was made morphoanatomical, ash and umidity content analysis, as well as the extracts and fractions in a Soxhlet apparatus. In addition, from the leaves, it was extracted the essential oils by hydrodistillation, and its further characterization by CGMS. From the crude extracts and hexane fractions it was performed the quantification of total chlorophylls and carotenoids. These samples, together with the remaining fractions and essential oil were analysed for their antioxidant potential through the molibdenium complex formation, radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) reduction and lipid peroxidation methods. Moreover, samples were evaluated for their toxicity against Artemia salina, hemolytic capacity and phytotoxicity. Separation techniques were also applied, in order to isolate and/or identify compounds in the species. The results obtained highlighted morphoanatomical characteristics similar to the ones described previously for the species. The essential oil was extracted from two collections in distinct periods, revealing the presence of spathulenol as one of the major constituents in both samples. The antioxidant assays demonstrated a better activity for ethyl acetate and etanolic fraction from the stem bark, and a potential toxic activity for the essential oil and stem bark hexane fraction. Separation techniques applied lead to isolation the aminoacid 4-hydroxy-N-methylproline, from the stem bark remanescent fraction. From this, it is suggested that the study of O. disospyrifolia should be continued, since there is a possibility of applicability from its extracts, fractions, essential oil and possible isolated compounds in the pharmacology area. Keywords: Lauraceae. Essential oil. Spathulenol. 4-hydroxy-N-methylproline. Toxicity