Extrato seco de Mouriri pusa Gardner (Melastomataceae): preparação, quantificação de flavonoides, secagem por aspersão e citotoxicidade
Data
2019
Autores
Ravazoli, Marcio Roberto 
Orientadores
Andreo, Marcio Adriano
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Desde os tempos remotos, o homem precisou aprender a caçar, se vestir, se proteger e fez isso
mimetizando o comportamento dos animais, que o fazem por instinto. Com o ato de curar não
foi diferente. No Brasil, o uso de plantas bioativas é anterior ao período de colonização. Usadas
pelos indígenas, as espécies brasileiras bioativas da flora nativa tiveram seu registro com a
chegada dos jesuítas ao país, os quais, além da colonização destes povos, também atuavam
como boticários, médicos e enfermeiros. Atualmente, o cerrado brasileiro continua sendo um
berço de espécies vegetais com potencial para se tornarem fitoterápicos, incluindo a espécie
Mouriri pusa Gardner (Melastomataceae), utilizada popularmente como antiúlceras gástricas,
sobretudo nas regiões norte e nordeste. Esta atividade deve estar relacionada à presença de
taninos e flavonoides nos extratos alcoólicos e aquosos. Neste contexto, o objetivo deste
trabalho foi realizar a obtenção de extrato seco e padronizado a partir das folhas de M. pusa por
aspersão, utilizando os flavonoides quercetina e miricetina como marcadores. A caracterização
da matéria-prima quanto a granulometria, teor de cinzas e teor de umidade, seguiu
recomendações da Farmacopeia Brasileira 5ª edição. O extrato líquido foi obtido por percolação
e avaliado quanto ao seu teor alcoólico, densidade relativa, teor de sólidos totais e teor de
flavonoides. O processo de secagem por aspersão foi desenvolvido e avaliado por meio de
planejamento experimental do tipo fatorial completo 23
. Os fatores avaliados foram fluxo de
alimentação, concentração de adsorvente (Aerosil®
200) e temperatura de entrada, avaliandose o rendimento, umidade e o teor de flavonoides como resposta. Em seguida, a condição que
proporcionou maior concentração de flavonoides, menor umidade e maior rendimento, foi
utilizada para a obtenção do extrato seco. O extrato obtido passou por avaliação de umidade,
rendimento, teor de flavonoides, tamanho e forma das partículas, área superficial e também por
ensaio de citotoxicidade. As folhas secas e moídas de M. pusa, classificadas como pó fino a
finíssimo, foram analisadas apresentaram em média de 8,54 % de umidade e 4,46 % de cinzas
totais. O extrato líquido apresentou teor alcoólico de 69%, densidade relativa de 0,896 g/mL,
concentração de sólidos totais de 11,5 mg/mL e concentração de flavonoides totais de 190,4
μg/mL, sendo 134,1 μg/mL de quercetina e 56,3 μg/mL de miricetina. O processo de secagem
foi otimizado, apresentando como melhores condições, a temperatura de entrada de 160°C, a
concentração de Aerosil®
200 de 50% e o fluxo de alimentação de 6 mL/min. Nestas condições,
o extrato apresentou teor de umidade 3,2%, rendimento de 78,4% e teor de flavonoides entre
0,5 e 1% (0,71%). A microscopia eletrônica de varredura demonstrou partículas esféricas com
variação de tamanho entre 3 e 15,1 µm. O extrato seco também apresentou uma área superficial
específica de 86.829 m²/g. O valor de IC50 obtido para citotoxicidade foi de 4.6 mg/mL, por
meio deste valor, foi possível estimar a DL50 de 2435.3 mg/kg. Portanto, o processo de secagem
se mostrou adequado, possibilitando a obtenção de um extrato seco com teor de 0,71% de
flavonoides, além de baixo teor de umidade e rendimento >75%.
From ancient times man had to learn to hunt, dress, protect himself by mimicking the animals behavior, which they did on instinct. With the act of healing it was no different. In Brazil, the use of bioactive plants predates the colonization period. Used by the natives, the registration of bioactive Brazilian native species had been made after the Jesuits arrival, who, besides the colonization of these towns, also acted as apothecaries, doctors and nurses. Currently, Brazilian cerrado remains as medicinal plant species crib, including Mouriri pusa Gardner (Melastomataceae), popularly used as antiulcer gastric, mainly in the north and northeast. This activity probably is related to the tannins and flavonoids presence in alcoholic and aqueous extracts. In this context, the objective of this work was to obtain dry and standardized extracts from the M. pusa leaves, applying the spray drying technique, using the flavonoids quercetin and myricetin as chemical markers. The characterization of the raw material as granulometry, ash and moisture content, followed the Brazilian Pharmacopoeia fifth edition recommendation. The liquid extract was obtained by percolation and evaluated for its alcohol content, relative density, total solids content and flavonoid content. The spray-drying process was developed and evaluated through experimental design, full factorial 23 type. The factors evaluated were feed flow, adsorbent concentration (Aerosil® 200) and inlet temperature, and yield, moisture and flavonoid content were the responses avaliated.Then, the condition that provided higher concentration of flavonoids, lower humidity and higher yield, was used to obtain the dry extract. The extract obtained underwent evaluation of humidity, yield, flavonoid content, particle size and shape, surface area and also by cytotoxicity test. The dried and ground leaves of M. pusa, classified as fine to very thin powder, were analyzed with an average of 8.54% of humidity and 4.46% of total ashes. The liquid extract had a 69% alcohol content, a relative density of 0.896 g / mL, a total solids concentration of 11.5 mg / mL and a total flavonoid concentration between 190.4 μg / mL to 134.1 μg / mL by quercetin and 56.3 μg / mL by myricetin. The drying process was optimized, presenting the best conditions, the inlet temperature of 160 ° C, the Aerosil® 200 concentration of 50% and the feed flow of 6 mL / min. Under these conditions, the extract had a moisture content of 3.2%, yield of 78.4% and flavonoid content between 0.5 to 1% (0.71%). Scanning electron microscopy showed spherical particles varying in size from 3 to 15.1 μm. The dry extract also had a specific surface area of 86,829 m²/g. The IC50 value obtained by cytotoxicity was 4.6 mg / mL. Using this value, it was possible to estimate the LD50 2435.3 mg / kg. Therefore, the drying process was adequate, making it possible to obtain a dry extract with 0.71% of flavonoids content, in addition a low moisture content and a yield >75%.
From ancient times man had to learn to hunt, dress, protect himself by mimicking the animals behavior, which they did on instinct. With the act of healing it was no different. In Brazil, the use of bioactive plants predates the colonization period. Used by the natives, the registration of bioactive Brazilian native species had been made after the Jesuits arrival, who, besides the colonization of these towns, also acted as apothecaries, doctors and nurses. Currently, Brazilian cerrado remains as medicinal plant species crib, including Mouriri pusa Gardner (Melastomataceae), popularly used as antiulcer gastric, mainly in the north and northeast. This activity probably is related to the tannins and flavonoids presence in alcoholic and aqueous extracts. In this context, the objective of this work was to obtain dry and standardized extracts from the M. pusa leaves, applying the spray drying technique, using the flavonoids quercetin and myricetin as chemical markers. The characterization of the raw material as granulometry, ash and moisture content, followed the Brazilian Pharmacopoeia fifth edition recommendation. The liquid extract was obtained by percolation and evaluated for its alcohol content, relative density, total solids content and flavonoid content. The spray-drying process was developed and evaluated through experimental design, full factorial 23 type. The factors evaluated were feed flow, adsorbent concentration (Aerosil® 200) and inlet temperature, and yield, moisture and flavonoid content were the responses avaliated.Then, the condition that provided higher concentration of flavonoids, lower humidity and higher yield, was used to obtain the dry extract. The extract obtained underwent evaluation of humidity, yield, flavonoid content, particle size and shape, surface area and also by cytotoxicity test. The dried and ground leaves of M. pusa, classified as fine to very thin powder, were analyzed with an average of 8.54% of humidity and 4.46% of total ashes. The liquid extract had a 69% alcohol content, a relative density of 0.896 g / mL, a total solids concentration of 11.5 mg / mL and a total flavonoid concentration between 190.4 μg / mL to 134.1 μg / mL by quercetin and 56.3 μg / mL by myricetin. The drying process was optimized, presenting the best conditions, the inlet temperature of 160 ° C, the Aerosil® 200 concentration of 50% and the feed flow of 6 mL / min. Under these conditions, the extract had a moisture content of 3.2%, yield of 78.4% and flavonoid content between 0.5 to 1% (0.71%). Scanning electron microscopy showed spherical particles varying in size from 3 to 15.1 μm. The dry extract also had a specific surface area of 86,829 m²/g. The IC50 value obtained by cytotoxicity was 4.6 mg / mL. Using this value, it was possible to estimate the LD50 2435.3 mg / kg. Therefore, the drying process was adequate, making it possible to obtain a dry extract with 0.71% of flavonoids content, in addition a low moisture content and a yield >75%.