Anti-inflammatory effects and acute oral toxicity of Copaifera spp. essential oil-loaded nanoemulsion / Efectos antiinflamatorios y toxicidad oral aguda de nanoemulsión cargada de aceite esencial de Copaifera spp

Copaifera spp. essential oil (EOC) was extracted by hydrodistillation of Copaifera oleoresin (COR). The EOC was characterized by GC/MS and a novel EOC-loaded nanoemulsion was developed to enhance the EOC solubility and to evaluate its utility as antinflammatory. EOC contain 14 volatile compounds (including ß-caryophyllene: 51.52%) having a required HLB of 11. The Surfactant: EOC: Water ratio of 13:15:75 (%, w:w:w) produced the optimal formulation (particle size: 94.47 nm). The EOC-loaded nanoemulsion presented a pseudoplastic/thixotropic behavior with excellent shelf stability for 6 months. The anti-inflammatory effect of the nanoemulsion was more potent than that of the EOC, and statistically equal to diclofenac (50 mg/kg). The EOC-loaded nanoemulsion showed no oral acute toxicity (in mice) at 2000 mg/kg; hence, it is considered a nontoxic product. The development of the EOC-loaded nanoemulsion added value to both the COR and the EOC by providinga suitable formulation that could be used as an anti-inflammatory product.

El aceite esencial (EOC) fue extraído por hidrodestilación de oleoresina de Copaifera spp. El EOC fue caracterizado químicamente por GC/MS. Se formuló una nanoemulsión con EOC para mejorar la solubilidad del EOC y evaluar su utilidad como antiinflamatorio. El EOC contiene 14 compuestos volátiles (incluido el ß-cariofileno: 51,52%) con un HLB requerido de 11. La relación Tensioactivo: EOC: Agua de 13:15:75 (%, p:p:p) produjo la formulación óptima (tamaño de partícula: 94,47 nm).. La nanoemulsión cargada con EOC presentó un comportamiento pseudoplástico/tixotrópico con una excelente estabilidad en almacenamiento durante 6 meses. El efecto antiinflamatorio de la nanoemulsión fue más potente que el del EOC y estadísticamente igual al diclofenaco (50 mg/kg). La nanoemulsión cargada con COE no mostró toxicidad aguda oral (en ratones) a 2000 mg/kg; por lo tanto, se considera un producto no tóxico. El desarrollo de la nanoemulsión cargada con EOC agregó valor tanto al COR como al EOC al proporcionar una formulación adecuada que podría usarse como un producto antiinflamatorio.

Cassia oleoresin Mediated Synthesis of Magnesium Oxide Nanoparticles and Brine Shrimp Lethality Assay

Cassia oleoresin is an extract isolated from dried barks of Cinnamomum cassia Blume (family Lauracea). The plant has been reported to have anti-diabetic, anti-oxidant, anti-hypertriglyceridemic effect, mainly due to its phytochemical constituents such as phenolic and volatile compounds. Cinnamon also helps in arthritis, fibromyalgia and psoriasis. The aim of this study was to prepare magnesium oxide nanoparticles using Cassia oleoresin and to evaluate the cytotoxic effect on Brine shrimp. The magnesium oxide nanoparticle was prepared from magnesium chloride and Cassia oleoresin and was confirmed by UV- Visible Spectroscopy and morphology was confirmed by TEM. Brine shrimps lethality bioassay was carried out to investigate the cytotoxicity of Cassia oleoresin mediated magnesium oxide nanoparticles. Ten brine shrimp nauplii were placed in each well of the Eliza plate and filled with 5 μL ,10 μL ,15 μL ,20 μL ,25 μL of Cassia oleoresin mediated magnesium oxide nanoparticles After 24 hours of incubation, the wells were observed and the number of surviving brine shrimp nauplii were counted to assess the cytotoxicity. The UV -Visible spectroscopy showed a peak at 400 peak and TEM analysis showed a particle size of 70 nm. After 24 hours incubation of the brine shrimps in the nanoparticle solution, all 10 brine shrimps survived in 5μL and 10 μL concentrations. 3 brine shrimps nauplii survived in 15μL conc. 1 brine shrimp nauplii survived in 20μL and 25μL concentrations each. Within the limits of this study it can be concluded that at low concentrations the prepared nanoparticle was safe and may be used for biomedical application.

Antibacterial Activity of White Pepper Oleoresin Mediated Silver Nanoparticles against Oral Pathogens.

Plant mediated biological synthesis of silver nanoparticles is gaining importance due to its eco friendliness and simplicity. Different types of nanoparticles such as metal, metal oxide, metal sulphide and polymer nanoparticles are used in different types of applications in the fields of electronics, communications, electrical, mechanical, medical, dental etc. Aim of the present study was to evaluate the antibacterial activity of white pepper oleoresin mediated silver nanoparticles against oral pathogens. MethodsIn the present investigation, white pepper mediated synthesis of silver nanoparticles was carried out by the addition of silver nitrate with prepared white pepper oleoresin solution. White pepper oleoresin mediated silver nanoparticles were initially confirmed by colour changes and confirmed by UV- Visible spectroscopy. Anti-bacterial activity of the synthesised silver nanoparticles was carried out using agar well diffusion method. Different concentrations of silver nanoparticles were tested against different oral pathogens such as Staphylococcus aureus, Streptococcus mutans and Pseudomonas sp. ResultsThe solution containing white pepper oleoresin and silver nitrate solution showed a brown colour confirming the formation silver nanoparticles and peak at 350 nm revealed the silver nanoparticles surface plasmon resonance. The white pepper oleoresin mediated silver nanoparticles showed good antibacterial activity against all tested microorganisms and the maximum activity was found with Pseudomonas spp. followed by Staphylococcus aureus, and Streptococcus mutans. ConclusionsGreen synthesis of silver nanoparticles using white pepper oleoresin was of low cost and was convenient to carry out. Hence, white pepper oleoresin mediated silver nanoparticles may be used for the control of oral pathogens.

Study of the in vitro release profile of sesquiterpenes from a vaginal cream containing Copaifera duckei Dwyer (Fabaceae) oleoresin.

Copaiba trees and their oleoresin is an important natural product used as an anti-inflammatory and antimicrobial drug, in particular for gynecological infections. The in vitro release profile of sesquiterpenes in a vaginal cream containing copaiba (CVC – copaiba vaginal cream) oleoresin in buffered solutions at different pHs was evaluated in the present study. The dissolution test revealed that trans-caryophyllene (TC) was constantly released until the final test time. The maximum peaks were found at a concentration of 478.79 nL/mL for the buffer at pH 5 and at a concentration of 475.99 nL/mL for the buffer at pH 6, with a percentage release of 50.52% and 50.21%, respectively. Likewise, the caryophyllene oxide (CO) was released into the buffers at pHs 5 and 6, with maximum release peaks at 60 minutes at a percentage of 31.8% and 33.5% corresponding to concentrations of 2.779 and 2.925 μg/mL, respectively. The release and dissolution profiles of the sesquiterpene compounds indicated that a pH of 5-6 in the dissolution media is adequate for release of these markers of antimicrobial activity in Copaifera duckei Dwyer. Therefore, these conditions possibly favor the therapeutic action of CVC.

Extraction of ginger oleoresin from Zingiberis Rhizoma Recens and inclusion of ginger oleoresin with β-cyclodextrin / 中国药学杂志

OBJECTIVE: To optimize the extraction and inclusion processes of ginger oleoresin from Zingiberis Rhizoma Recens. METHODS: Supercritical carbon dioxide extraction was selected to extract ginger oleoresin from Zingiberis Rhizoma Recens. The extraction condition was optimized by orthogonal experiment. The inclusion method, the ratio of ginger oleoresin to β-cyclodextrin and inclusion time were studied as factors of inclusion process. RESULTS: The optimized extraction technology was as follows: extraction temperature was 50℃, extraction pressure was 25 MPa, separation pressure was 9 MPa and extraction time was 90 min. The optimal inclusion method was triturating inclusion, with the ratio of ginger oleoresin to β-cyclodextrin of 1:8 and inclusion time of 60 min. CONCLUSION: The optimized technology of extraction and inclusion is stable and feasible, and can be used for the extraction and inclusion of ginger oleoresin.

Development of a larvicidal nanoemulsion with Copaiba (Copaifera duckei) oleoresin

Copaiba (Copaifera duckei Dwyer, Fabaceae) oleoresin is an important Amazonian raw material. Despite its insecticidal potential, poor water solubility remains a challenge for the development of effective and viable products. Nanotechnology has emerged as a promising area to solve this problem, especially oil-in-water nanoemulsions. On this context, the aim of the present study was to develop oil-in-water nanoemulsions using copaiba oleoresin dispersed through a high internal phase; and evaluate its potential insecticidal action against Aedes aegypti larvae. Overall, 31 formulations were prepared, ranging from 11.5 ± 0.2 to 257.3 ± 4.1 nm after one day of manipulation. Some of them reached small mean droplet sizes (< 200 nm) and allowed achievement of a nanoemulsion region. The formulation consisted of 5% (w/w) of copaiba oil, 5% (w/w) of surfactant and 90% (w/w) of water, which presented mean droplet size of 145.2 ±0.9 nm and polidispersity of 0.378 ± 0.009 after one day of manipulation, and these were evaluated for larvicidal potential. According to mortality level (250 ppm - 93.3 after 48 h), this nanoemulsion was classified as a promising insecticidal agent against Aedes aegypti larvae. The present study allowed the development of low-cost ecofriendly green natural-based nanoformulations with potential larvicidal activity, using a nanobiotechnology approach.

Antimicrobial activity of copaíba (Copaifera langsdorffii) oleoresin on bacteria of clinical significance in cutaneous wounds / Atividade antimicrobiana do óleo-resina de copaíba (Copaifera langsdorffii) em bactérias de significância clínica em úlceras cutâneas

The present study was designed to evaluate the in vitro antimicrobial activity of Copaifera langsdorffii oleoresin, which has been used in folk medicine as an anti-inflammatory, antibacterial, healing among others. The oleoresin was tested against Gram-positive (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria related to infections in cutaneous wounds. Antimicrobial activity was determined by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Copaiba oleoresin showed antimicrobial activity only against the Gram-positive bacteria with MIC of 200 µg/mL, 400 µg/mL and 1100 µg/mL for S. aureus, S. pyogenes and E. faecalis, respectively. MBC values were the same as MIC for S. aureus and S. pyogenes and for E. faecalis it was 1200 µg/mL. Considering that infection significantly impairs the wound healing process, we believe that the use of copaiba oleoresin as a component of a topical formulation could be a valuable adjunct in the treatment of infected wounds, mainly in the case of wounds infected by Gram-positive microorganisms.

Este trabalho avaliou a atividade antimicrobiana in vitro do óleo-resina da Copaifera langsdorffii, o qual vem sendo utilizado há muitos anos na medicina tradicional popular, principalmente devido às suas propriedades antiinflamatórias, antibacterianas, cicatrizante entre outras. O óleo-resina foi testado em bactérias Gram-positivas (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) e Gram-negativas (Pseudomonas aeruginosa e Escherichia coli) relacionadas com infecções de úlceras cutâneas. A atividade antimicrobiana foi determinada pelos testes da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). O óleo-resina apresentou atividade antimicrobiana in vitro apenas para as bactérias Gram-positivas, com valores de CIM de 200 µg/mL, 400 µg/mL e 1100 µg/mL para S. aureus, S. pyogenes e E. faecalis, respectivamente. Os valores de CBM foram os mesmos que os valores de MIC para S. aureus e S. pyogenes. O valor de CBM para o microrganismo E. faecalis foi de 1200 µg/mL. Considerando que a presença de infecção significativamente impede o processo normal de cicatrização de úlceras cutâneas, acreditamos que o óleo-resina de copaíba, utilizado como componente de formulações tópicas, poderia ser um adjunto importante no tratamento de úlceras cutâneas infectadas, principalmente nos casos de infecção por microrganismos Gram-positivos.

Formulation of a Functional Fat Spread Stabilised by Natural Antioxidants and Emulsifiers

Introduction: Essential fatty acids (EFAs) play a vital role in the human body and need to be taken through a regular diet. EFAs are susceptible to autoxidation, hence the stability of the EFAs and their products is a matter of concern. Methods: Margarine containing sunflower oil as a carrier of EFAs was prepared and the effects of water content, incorporation of EFAs, emulsifiers and antioxidants on the physical properties of margarine, that is, slipping point, dropping point and spreadability were studied. The oxidative stability of the formulated margarine was also evaluated after incorporation of EFAs and antioxidants. Results: The incorporation of EFAs in the form of sunflower oil resulted in improved physical properties especially spreadability. The study revealed that up to 45% sunflower oil can be incorporated using glycerol monostearate as an emulsifier with total fat to water ratio of 85:15. Lecithin imparted better spreadability and grainy structure but is known to be susceptible to microbial attack. The capsicum oleoresin showed good activity as an antioxidant. Further addition of kalonji seeds ethanol extract (KEE) as well as curcuminoids resulted in improved spreadability but showed a decrease in oxidation stability. Conclusion: A stable and nutritional margarine was developed with the addition of natural antioxidants. Consumers can avail the benefits of both the EFAs and natural antioxidants in the margarine.

Avaliação do efeito do óleo-resina de copaíba (Copaifera s) na proliferação celular in vitro / Effect of copaiba oleoresin (Copaifera sp) on the in vitro cellular proliferation

O óleo de copaíba é uma resina extraída pela perfuração do tronco de árvores do gênero Copaifera sp., usado como cicatrizante em várias regiões do Brasil. São comprovados seus efeitos antimicrobianos e antiinflamatórios, sem relatos de atividade sobre a proliferação celular. No presente trabalho foi observada a dinâmica da proliferação de células MDBK (Madin Darby Bovine Kidney) mantidas em meio de cultivo adicionado de diferentes concentrações do óleo de copaíba, utilizando-se como controle células mantidas em meio sem adição do oleoresina (grupo M) e células no meio com aplicação do solvente tween 80, na diluição 10-3 (grupo MT). Diluições decimais de 10-1 até 10-3 mostraram-se tóxicas e, portanto, os estudos de proliferação partiram da diluição 10-4 até 10-7. Observou-se que houve crescimento mais acelerado em todos os grupos adicionados do óleo-resina nas primeiras 24 horas, com destaque para a diluição 10-5, que teve sua taxa de proliferação 5,47 vezes maior que a do grupo M. Concluiu-se que o óleo-resina de copaíba se mostrou estimulante da multiplicação celular, o que pode ser um dos mecanismos de seu efeito positivo sobre a cicatrização, somado àqueles previamente comprovados na literatura.

Copaiba oleoresin is extracted from the trunk of Copaifera sp genus trees and used for treating wounds in several regions of Brazil. Its antimicrobial and anti-inflammatory effects have been proved, however there are no reports of activity on cell growth. Proliferation of MDBK (Madin Darby Bovine Kidney) cells was evaluated under the influence of different concentrations of Copaiba oleoresin. Control groups consisted of cells in medium without addition of oleoresin (M group) and cells in the medium with application of the solvent Tween 80 at dilution 10-3 (TM group). Decimal dilutions of 10-1 to 10-3 were shown to be toxic and, therefore, the proliferation studies were conducted from dilution 10-4 to 10-7. Cell growth was faster in all groups that received the Copaiba oleoresin dilutions in the first 24 hours, specially the 10-5 dilution group, which proliferation rate was 5,47 higher than that of M group. It was concluded that Copaiba oleoresin stimulates cell multiplication, which may be one mechanism of its positive effect on wound healing, in association with those previously known.

Composición de las oleorresinas de dos variedades de ají picante (habanero y tabasco) obtenidas mediante lixiviación con solventes orgánicos / Composition of oleoresins from two kinds of chili pepper (habanero and tabasco) obtained by lixiviation with organic solvents

Introducción. En el desarrollo de nuevos procesos e ingredientes para la industria alimentaria, las técnicas de extracción permiten obtener principios activos a partir de productos naturales, aptos para el consumo humano y fuertemente relacionados con la tendencia hacia una alimentación más saludable y libre de contaminantes. Las oleorresinas de especies vegetales permiten el manejo simultáneo de componentes pigmentantes y aromatizantes como en el caso de los pimientos, para los cuales la extracción con solventes orgánicos es una práctica común que debe refinarse de tal modo que se empleen sustancias de extracción seguras y aprobadas para el consumo humano. Objetivo. Comparar la composición de la oleorresina de dos variedades de ají picante (tabasco y habanero) extraída con dos solventes diferentes (hexano y acetato de etilo). Materiales y métodos. Se tomó material seco de dos variedades de ají picante (tabasco y habanero) y se sometieron a extracción con dos solventes orgánicos diferentes (hexano y acetato de etilo). Las oleorresinas resultantes fueron concentradas para calcular el rendimiento en extracción y luego fueron analizadas en un HPLC para determinar su contenido en capsaicina y dihidrocapsaicina. Resultados. Se encontró que el acetato de etilo es un mejor solvente que el hexano, pues presenta un porcentaje general de extracción del 11,23% frente a 7,14%. En la caracterización por HPLC se encontraron concentraciones aceptables de los principios pungentes (capsaicina y dihidrocapsaicina) las cuales corresponden con los valores teóricos en los cuales el ají habanero presenta entre 150.000 y 325.000 Unidades Scoville y tabasco presenta entre 50.000 y 100.000 Unidades Scoville. Conclusión. En un proceso de extracción con solventes sobre ají picante, se obtiene un mejor rendimiento en extracción empleando acetato de etilo como solvente y una mayor concentración de capsaicina y dihidrocapsaicina empleando ají habanero.

Introduction. In the development of new processes and ingredients for food industry, extraction techniques allow us to obtain active principles from natural products, good for human consuming and strongly related to the tendency towards a healthier and free of contaminants feeding. Oleoresins from vegetables allow us to work with dye and aromatic components (as in the case of peppers, for which, extraction with organic solvents is a common practice that must be refined in such a way that only safe and apt for human consumption substances are used). Objective. To compare the composition of oleoresin from two kinds of chili pepper (Tabasco and habanero) extracted with two different solvents (hexane and ethyl acetate). Materials and methods. Dry material from both kinds of chili pepper were taken and submitted to extraction with two different organic solvents (hexane and ethyl acetate). The oleoresins that resulted from this procedure were concentrated to calculate their performance in extraction and then analyzed in a HPLC to determine their contents of capsaicin and dihydrocapsaicin. Results. We found that ethyl acetate is a better solvent than hexane; it shows a general percentage of extraction of 11.23% compared to a 7.14%. In the characterization with the HPLC we found acceptable concentrations of the pungent principles (capsaicine and dihydrocapsaicin), corresponding to the theoretical values in which habanero pepper represents a value between 150000 and 325000 Scoville units and the Tabasco, between 50000 and 10000 Scoville units. Conclusion. In an extraction process for chili peppers, a better extraction performance is obtained by using ethyl acetate as solvent and a better concentration of capsaicin and dihydrocapsaicin by using habanero chili pepper.

Evaluation of ginger (Zingiber officinale Roscoe) bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ in-vitro

The potential ability of ginger bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ was investigated using dual tagging FITC and PE of monoclonal antibody anti-human with its fluorescence measured by flow cytometer. Oleoresin was extracted using sinkhole distillation technique. Its components namely, gingerol in fraction-1, shogaol in fraction 2 and zingeron in fraction-3 were separated by column vacuum chromatography method. The doses of oleoresin, gingerol, shogaol, and zingeron tested were 50, 100,150, 200, and 250 μg/ml. Lymphocytes (2x106 cell/ml) from human peripheral blood were isolated using ficoll density gradient technique, and cultured in the presence of the compounds in RPMI-1640 medium and phytohemaglutinin (PHA) mitogen for 96 h under normal conditions. Percentages of T-cell surface molecules (CD4+ and CD8+) were determined using dual-tagging FITC and PE fluorescents labeled on monoclonal antibody anti human. The fluorescence-labeled bands on the T-cell surface molecules were counted using flow cytometer. The experiment revealed that oleoresin and its three fractions increased the percentage of CD3+CD4+. The compound in fraction 3 of oleoresin at 200 μg/ml increased by the highest percentage of CD3+CD4+ of 9%, but slightly decreased the percentage of CD3+CD8+. These ginger bioactive compounds increased the ratio of CD3+CD4:CD3+CD8+ T-cells with the highest increment of 30% from effects of 200 μg/ml fraction 3 of oleoresin. This in vitro finding revealed that ginger bioactive compounds potentially increased cellular and humoral immune response. Further clinical studies are needed to confirm the benefits of these ginger bioactive compounds as a potential functional food for testing on HIV infected patients.

THE THERAPEUTIC EFFECT OF GINGER OLEORESINS ON HYPERLIPIDEMIA AND PLATELET AGGREGATION / 营养学报

Objective: To study the effects of ginger oleoresins(GO) on blood lipids in hyperlipidemic mice and the platelet aggregation in rats.Methods: The rat platelet aggregation was induced by ADP and AA in vitro, and the inhibition by GO was tested by light transmission. The animal model of hyperlipidemia was establised by feeding Kunmimg mice with high lipid feed stuff. The effects of GO on serum lipid and hepatic MAD in hyperlipidemic mice was investigated. Results: GO could effectively inhibit rat platelet aggregation induced by ADP and AA in vitro, which were 58.9%,97.5% respectively, and in dose dependant manner. Besides, GO could also significantly decrease serum TC, LDL, AS index and hepatic MDA in hyperlipidemic mice, which were 16.45%, 60.45%,37.30% and 54.27% respectively, and showed no side-effect on liver. Conclusion: Long-term consumption of GO can effectively modulate blood lipids, prevent thrombosis and AS.

RAPID DETERMINATION OF TOTAL VITAMIN E IN LYCOPENE OLEORESIN BY SOLID PHASE EXTRACTION AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY / 营养学报

Objective:To establish a rapid determination method of total VE in lycopene oleoresin.Method:By eliminating saponifiable matters and strong polar materials from the sample,VE was concentrated efficiently by PRP-6 SPE with 90% ethanol/water and 100% ethanol as elution solvents consecutively.The removal of polar materials was satisfactory.VE was determined by HPLC.Results:The linear regression was carried out,the relative coefficient r=0.9998 and 99.85～99.95 % recovery of VE were achieved.Conclusion:A simple and rapid VE determination method with good sensitivity is developed.