ABSTRACT
AbstractPassiflora alata Curtis, Passifloraceae, is a liana popularly known in Brazil as ‘maracujá-doce’ that has been used for treating different illnesses. Its leaves are described in the Brazilian Pharmacopoeia, but the gastroprotective activity has never been investigated. In the present study a freeze-dried crude 60% ethanol–water extract of P. alata aerial parts was prepared. Total flavonoid content, expressed as vitexin, was 0.67% ± 0.01. The hemolytic activity was 32 units for P. alata, using Saponin (Merck®) as reference. P. alata presented EC50 of 1061.2 ± 8.5 µg/ml in the 2,2-diphenyl-1-picryhydrazyl assay and 1076 ± 85 µmol Trolox/g in the Oxygen Radical Absorbance Capacity assay. P. alata, its solvent fractions and a P. alatananopreparation were investigated for gastroprotective activity. The test samples exhibited gastroprotective activity on HCl/ethanol induced gastric mucosal lesions in rats. P. alata at doses of 100, 200 and 400 mg/kg, using the necrotizing agent at 150 mmol/l, inhibited 100% of ulcer formation (compared to the negative control), while lansoprazole (30 mg/kg) 77%. When tested against a more concentrated necrotizing agent (300 mmol/l), fractions of P. alata at 100 mg/kg reduced 57% (n-hexane), 34% (ethyl acetate) and 72% (aqueous fraction) the ulcer formation. In this assay, lansoprazole (30 mg/kg) inhibited 47%. When encapsulated, P. alata inhibited ulcer formation at 55%, 94% and 90% for dosages of 25, 50 and 100 mg/kg. These results suggest the potential use of P. alata as a gastroprotective herbal medicine.
ABSTRACT
No Brasil, os produtos fitoterápicos são considerados medicamentos, sendo necessário o estabelecimento de estudos que assegurem a manutenção dos requisitos de qualidade durante o processamento e o armazenamento. Testes de dissolução podem ser empregados para se estimar a biodisponibilidade de um fármaco, sendo uma análise rotineira no desenvolvimento e controle de qualidade de medicamentos alopáticos. A determinação do perfil de dissolução de fitoterápicos também pode ser um importante critério para avaliação da sua qualidade lote-a-lote, bem como para os estudos de desenvolvimento e de estabilidade. O objetivo deste trabalho foi investigar a influência dos métodos de secagem e da condição de armazenagem sobre os perfis de dissolução dos flavonoides totais de extratos secos de duas plantas medicinais bastante difundidas no Brasil, a Bauhinia forficata e a Passiflora alata. Os extratos secos foram produzidos pelo processo de secagem em leito de jorro e em spray drying, sendo submetidos a condições de armazenagem aceleradas (temperatura de 40 ± 2ºC e umidade relativa de 75 ± 5%, por um período de 90 dias). Os perfis de dissolução foram obtidos para amostras de extratos secos antes e após o período de armazenamento. O teor de flavonoides totais foi quantificado por espectrofotometria. Os extratos secos de B. forficata e P. alata apresentaram adequada liberação de flavonoides nos ensaios de dissolução. Os extratos secos de Passiflora alata apresentaram completa dissolução dos flavonoides, 92% e 98% dos teores originais após 60 minutos de ensaio, respectivamente para o extrato seco em leito de jorro e em spray drying.
In Brazil, most of the herbal medicinal products are considered as medicine. Therefore, it is necessary the establishment of tests to guarantee the maintenance of quality requirements during their processing and storage. The dissolution test is used to estimate the bioavailability of drugs and is routinely used in the development and the quality control of allopathic medicines. The determination of the dissolution profile of herbal products can also be an important criterion for assessing the batch-to-batch quality as well as for studies of product development and stability. This work aimed to investigate the dissolution profiles of dried extracts of two medicinal plants widely used in Brazil, the Bauhinia forficata and Passiflora alata, by assessing the effect of the drying methods and storage condition on the release of the total flavonoid contents. Spouted bed and spray drying were the processes used for the production of the dried extracts. The products were subjected to accelerated storage conditions (temperature of 40 ± 2ºC and relative humidity of 75 ± 5%, for 90 days). The dissolution profiles of the dried extracts, before and after storage, were determined. The concentration of total flavonoids was quantified by spectrophotometry. Adequate dissolution profiles of flavonoids from B. forficata and P. alata were obtained for all the dried extracts produced. The dried extracts of Passiflora alata showed the complete dissolution of flavonoids in the dissolution media investigated, respectively 92% and 98% of flavonoids present in the dried extracts in spouted bed and spray drying after 60 minutes of the dissolution testing.
Subject(s)
Plant Extracts/analysis , Passiflora/classification , Bauhinia/classification , Dissolution/analysis , Product Storage , Phytotherapeutic DrugsABSTRACT
The Passifloraceae family is extensively used in native Brazilian folk medicine to treat a wide variety of diseases. The problem of flavonoid extraction from Passiflora was treated by application of design of experiments (DOE), as an experiment with mixture including one categorical process variable. The components of the binary mixture were: ethanol (component A) and water (component B); the categorical process variable: extraction method (factor C) was varied at two levels: (+1) maceration and (-1) percolation. ANOVA suggested a cubic model for P. edulis extraction and a quadratic model for P. alata.These results indicate that the proportion of components A and B in the mixture is the main factor involved in significantly increasing flavonoid extraction. In regard to the extraction methods, no important differences were observed, which indicates that these two traditional extraction methods could be effectively used to extract flavonoids from both medicinal plants. The evaluation of antioxidant activity of the extract by ORAC method showed that P. edulis displays twice as much antioxidant activity as P. alata. Considering that maceration is a simple, rapid and environmentally friendly extraction method, in this study, the optimized conditions for flavonoid extraction from these Passiflora species is maceration with 75% ethanol for P. edulis and 50% ethanol for P. alata.
ABSTRACT
This review describes botanical, chemical, pharmacological and phytotechnological properties of Passiflora alata Curtis, with emphasis on analytical methods. Native to Brazil, P. alata is featured in several pharmaceutical preparations registered by the regulatory agency ANVISA. In four of the five editions of the Brazilian Pharmacopoeia, P. alata leaves have been included under the designation of maracujá or maracujá doce (sweet passion fruit). This species is cited as Passiflora alata Curtis and was originally published in 1788 in the Botanical Magazine. In the last decade, phytochemical and pharmacological studies of Brazilian researchers related to this plant have increased. Despite several studies, the substances responsible for the anxiolytic, sedative, antioxidative and antiulcer activities attributed to passifloras remain unknown. Analytical methods for the quantification of markers are being developed in order to improve quality control analysis and to better understand the relationship between chemical markers and their pharmacological effects. Those methods include spectrophotometry, high performance thin-layer chromatography (HPTLC) and high performance liquid chromatography (HPLC). There is also great interest in the technological processes involved in the production of dry extracts of this native medicinal plant with the primary goal of assuring the quality of phytopharmaceutical products.
En esta revisión se describen las características botánicas, químicas, farmacológicas y fitotecnologicas de Passiflora alata Curtis, con énfasis en los métodos analíticos. Originaria de Brasil, varias preparaciones farmacéuticas de P. alata son registradas por el organismo regulador ANVISA. En tres de las cinco ediciones de la Farmacopea Brasileña, las hojas de P. alata se han incluido bajo la denominación de "maracuyá" (fruta de la pasión). Esta especie debe ser citada como Passiflora alata Curtis, publicada originalmente en 1788 en Botanical Magazine. En la última década, los estudios fitoquímicos y farmacológicos de investigadores brasileños relacionados con esta planta han aumentado substancialmente. A pesar de varios estudios, las sustancias responsables de las actividades ansiolítica, sedante, antioxidante y antiulcerosa atribuidas a la passiflora continúan siendo desconocidos. Los métodos analíticos para la cuantificación de los marcadores están siendo desarrollados con el fin de mejorar el análisis de control de calidad y para comprender mejor la relación entre marcadores químicos y sus efectos farmacológicos. Estos métodos incluyen espectrometría de alto rendimiento, cromatografía en capa fina (HPTLC) y la cromatografía líquida de alta resolución (HPLC). También existe un gran interés en los procesos tecnológicos envueltos en la producción de extractos secos con el objetivo principal de asegurar la calidad de los productos fitosanitarios.
Subject(s)
Plant Extracts/chemistry , Passiflora/chemistry , Plants, Medicinal/chemistry , Brazil , Chromatography, High Pressure Liquid , Chromatography, Thin LayerABSTRACT
O presente trabalho teve como objetivo fazer uma análise comparativa da qualidade de preparações extrativas obtidas de Passiflora alata Curtis, além de descrever esse processo. O extrato líquido foi obtido por refluxo utilizando água como solvente em proporção 1:10. A preparação líquida foi submetida à secagem em spray dryer utilizando aerosil® e fosfato de cálcio como adjuvantes tecnológicos. A eficiência de extração foi avaliada através da determinação do resíduo seco e do teor de flavonoides totais por técnica de espectrofotometria no UV. O extrato seco foi avaliado quanto à viabilidade tecnológica a partir da determinação das características de granulometria e da estabilidade em ambientes de umidade relativa controlada. O teor de flavonoides totais para o extrato seco apresentou redução quando comparado à solução extrativa, indicando possível influência do processo de secagem na composição do material. Os resultados de caracterização tecnológica demonstraram que o processo de secagem utilizado garante uma uniformidade de tamanho particular, embora seja observada uma tendência de higroscopicidade do material em ambientes de elevada umidade relativa. As técnicas empregadas se mostraram viáveis para uso em ensaios quali-quantitativos aplicados à determinação da qualidade dos extratos desenvolvidos.
This paper describes a process to prepare liquid and dried aqueous extracts from aerial parts of Passiflora alata Curtis (sweet passionfruit) and reports a comparative analysis of the extractive preparations obtained. The extractive solution was obtained by refluxing the plant material with water in the proportion 1:10 (w/v). The aqueous extract was dried in a spray-dryer, with Aerosil® and calcium phosphate as processing aids. The extraction efficiency was assessed by weighing the dry residue and assaying the total flavonoid content by UV spectrophotometry. The particle size and hygroscopicity of the spray-dried powder were determined in order to assess its technological viability. In the solid state, the total flavonoid content was reduced, relative to the aqueous extract, which indicates a possible influence of the drying process on its composition. The technological characterization demonstrated that this drying process leads to particle size uniformity, though a tendency to hygroscopicity can be observed under conditions of high relative humidity. The analytical techniques employed proved to be viable for qualitative and quantitative assays used for quality control of the extracts developed in this study.
Subject(s)
Flavonoids/pharmacology , Passiflora , Plants, MedicinalABSTRACT
Preliminary work on Passiflora alata leaves failed to detect harmane alkaloids using LC. The aim of this work was to investigate the production of harmane alkaloids through the cell culture of P. alata, inducing its precursor (L-tryptophan). The leaf explants presented satisfactory results after disinfection, and the callus formation was initiated in MS media with adequate quantities of phytohormones. Sixty days after inoculation, calli were inoculated in the optimized semi-solid MS media, with and without the addition of L-tryptophan (50, 100, 200 mg/L) and kept in standard conditions for 90 days. Calli were collected on days 6, 16, 26, 36, and 90, followed by acid-base extraction, and analysed by LC. The results showed an absence of harmane, harmin, harmol, harmalol, and harmaline. With L-tryptophan feeding, two peaks were detected, collected and analysed through positive mode electrospray [ESI(+)-MS] and sequential analysis in tandem ESI(+)-MS/MS. The spectra obtained were very similar, with a repetition of the more intense ions, and consecutive loss of 68 Da units, attributed to the heterocycle pyrazole. It appeared that this transformation was not related to any enzymatic pathway previously described for the plant from L-tryptophan, and the biosynthesis of β-carboline alkaloids in callus culture of P. alata were not observed in this work.
As folhas de varias espécies de Passiflora são utilizadas como ansioliticas e sedativas. Passiflora alata Curtis, Passifloraceae consta em três edições da farmacopéia brasileira, porem não há muitos estudos sobre sua composição química. No passado, enfatizava-se a ação conjunta de alcalóides e flavonóides. Em trabalho anterior, não foi detectada a presença de alcalóides harmanicos através de CLAE. Assim, decidiu-se investigar a produção dos mesmos através de cultivo celular, introduzindo seu precursor metabólico L-triptofano. Os explantes foliares apresentaram resultados satisfatorios para germinação apos assepsia, e a formação de calo foi iniciada em meio MS com quantidades adequadas de fitohormonios, previamente determinadas. Sessenta dias após a inoculação os calos foram repicados para meio semi-solido com e sem L-triptofano (50, 100, 200 mg/L), mantidos por 90 dias em condições padrão. Amostras foram coletadas com 6, 16, 26, 36, e 90 dias, realizada extração acido-base e o extrato analisado por CLAE. Os resultados mostraram a ausência de harmana, harmina, harmol, harmalol e harmalina. Dois picos presentes nas amostras com L-triptofano foram coletados e analisados através de espectrometria de massas, electrospray modo positiva [ESI(+)-MS] e analise em tandem ESI(+)-MS/MS. Os espectros correspondentes foram similares, mostrando a perda consecutiva de 68 Da, atribuídos ao pirazol. Este fato aponta para uma transformação não enzimática, não relacionada a uma biossintese previamente descrita para alcalóides β-carbolínicos.
ABSTRACT
O extrato fluido (EF), preparado segundo a Farmacopéia Brasileira, e sua fração aquosa (FA) obtidos de folhas de Passiflora alata foram administrados por via oral em camundongos. Seus efeitos comportamentais foram avaliados por modelos que detectam a atividade ansiolítica e antidepressiva de drogas, tais como: o labirinto em cruz elevado (LCE) e o teste da suspensão pela cauda (TSC). Efeitos sobre a atividade locomotora geral dos animais foram monitorados no campo aberto. Efeitos sedativos foram observados com EF (100 e 300 mg kg-1) e EA (100, 300 e 600 mg kg-1), caracterizados por uma diminuição do número de entradas nos braços fechados do LCE e uma diminuição no número de cruzamentos e levantamentos no campo aberto. No TSC, a administração de EF (100 mg kg-1) ou FA (100 e 300 mg kg-1) resultou em aumento do tempo de imobilidade. Esses resultados são relevantes, pois contribuem para validar o uso popular dessa planta.
The fluid extract (FE), prepared according to the Brazilian Pharmacopoea, obtained from the leaves of Passiflora alata and its aqueous fraction (AF), were administered by oral route to mice, and the behavioural effects were evaluated using animal models that detect anxiolytic and antidepressant activities, such as the elevated plus maze (EPM) and the tail suspension test (TST). Effects on general motor activity were monitored in the open . Sedative effects were observed with FE (100 and 300 mg kg-1) and AF (100, 300 and 600 mg kg-1) and were characterized by a decreased number of entries in the enclosed arms of the EPM and a decrease in the number of crossings and rearing in the open . In the TST, FE (100 mg kg-1) and AF (100 and 300 mg kg-1) induced an increase in the immobility time. These results are relevant because they contribute to validate the traditional use of this plant.
Subject(s)
Animals , Mice , Passiflora , Plant Extracts/therapeutic use , Phytotherapy , Plants, MedicinalABSTRACT
Este trabalho investigou o efeito do extrato combinado de Passiflora alata Dryander e Valeriana officinalis L. (EPV) em ratos submetidos aos testes do labirinto em cruz elevado (LCE) e campo aberto (TCA). Nenhum efeito foi detectado após o tratamento agudo ou repetido por 3 ou 7 dias com EPV (5, 10 or 20 mg/kg, gavagem) no LCE e TCA. Entretanto, ratos tratados por 15 dias com EPV (20 mg/kg) mostraram aumento na porcentagem de entradas e tempo gasto nos braços abertos no LCE, sem alterar a atividade locomotora no TCA, comparado ao controle. Diazepan (droga de referência, i.p.), aumentou os mesmos parâmetros analisados no LCE e OFT após o tratamento agudo ou por 15 dias. O tratamento agudo com 300 ou 600 mg/kg do EPV diminuiu significativamente a atividade locomotora no TCA. Estes resultados mostram que EPV produz efeito ansiolítico e sedativo, com ampla margem de segurança para o efeito ansiolítico.
This work investigated the effects of a combined extract of Passiflora alata Dryander and Valeriana officinalis L. (EPV) in rats under going elevated plus maze (EPM) and open- test (OFT). No effects were detected after acute or repeated (3 or 7-days) treatment with EPV (5, 10 or 20 mg/kg, by gavage), on the EPM or the OFT. However, rats treated for 15 day (20 mg/kg) with EPV showed increased percentage of entries and time spent in the open arms on the EPM without alter locomotor activity in the OFT compared to control group. Acute or a 15 day administration of diazepam (2 mg/kg, i.p.), increased the same parameters on the EPM and OFT. Acute treatment with 300 or 600 mg/kg of EPV, decreased the locomotor activity in the OFT. Results suggest anxiolytic and sedative effects for the EPV and reveal a wide dose range for the anxiolytic effect.