Extract from byproduct Psidium guajava standardized in ellagic acid: additivation of the in vitro photoprotective efficacy of a cosmetic formulation

ABSTRACT The fruits of guava, Psidium guajava L., Myrtaceae, are cultivated as food and used in agroindustries, generating byproducts or waste that represent environmental problems and require adequate destination. However, these byproducts present high levels of secondary metabolites and have been awakened interest regarding to its reusing. The extract was standardized in ellagic acid concentration by high performance liquid chromatography. The additivation capacity in the in vitro photoprotective efficacy of guava byproduct extract standardized in ellagic acid was verified as a result of its incorporation in cosmetic formulations, comparing it with a standard product. The extract presented synergy with the chemical UV filter (ethylhexyl methoxycinnamate), enhancing the solar protection factor of the phytocosmetic in 17.99%. Besides that, it was possible to show its antioxidant activity and the presence of secondary metabolites such as phenols and flavonoids. According to the results, it is possible to claim that the extract from the guava's agroindustrial byproducts present potential to be studied and reused, applying on the development of innovative products intended to the photoprotection care.

Quantification of catechin in the spray-dried extract of Pimenta pseudocaryophyllus

Abstract This paper describes the quantification of catechin in the spray-dried extract of Pimenta pseudocaryophyllus (Gomes) Landrum, Myrtaceae, citral chemotype using a validated HPLC-PDA method. The method employs a RP-18 column with acetonitrile:water-orthophosphoric acid 0.05% (gradient system) and UV detection at 210 nm. The method was demonstrated to be simple, sensitive, specific, linear, precise, accurate and robust. The response was linear over a range of 5-200 µg/ml (r > 0.999). The range of recoveries was 92.27-102.54%. The relative standard deviation values for intra- and inter-day precision studies were 4.30 and 3.78%, respectively. This assay can be readily utilized as quality control method for catechin in the dried extract of P. pseudocaryophyllus.

Box-Behnken experimental design for extraction of artemisinin from Artemisia annua and validation of the assay method

ABSTRACT Obtaining artemisinin and its derivatives is very costly, which limits access to low-income people. Some hydroalcoholic extract of Artemisia annua L., Asteraceae, which has shown comparable antimalarial activity could be an alternative to the purified compound, especially if the cultivars have higher content of artemisinin. The objective of this study was to evaluate the effects of the extraction parameters (ethanol graduation, previous shaking time in an ultrasound bath and drug/solvent ratio) on the yield of artemisinin in the liquid extract obtained by percolation from A. annua and then optimize the extraction efficiency of this compound. The Box-Behnken (33) factorial design was used in association with response surface methodology. The derivatization reaction of artemisinin was used in an analytical method which was submitted to validation, after reaching the specification of the selectivity, linearity, precision, accuracy and robustness. Detection and quantification limits were 1.3 and 4.0 µg/ml, respectively. The largest amount of this compound of interest was obtained without any ultrasound bath, with an ethanol graduation of 95% and a drug/solvent ratio of 2%. Drug/solvent ratio was the factor which most influenced extraction efficiency. The maximum range of artemisinin yield was 1.21%. Information obtained in this study can be used for future approaches to determining and extracting artemisinin from A. annua.

Ação do extrato etanólico da casca do pequi (Caryocar brasiliense) na cardiotoxicidade crônica induzida por doxorrubicina em ratos / Caryocar brasiliense shell ethanol extract in chronic cardiotoxicity induced by doxorubicin in rats

A doxorrubicina (DOX) é um quimioterápico utilizado no tratamento de neoplasias malignas, porém possui a cardiotoxicidade como efeito colateral. O objetivo deste trabalho foi verificar quanto à ação do extrato etanólico da casca do pequi (Caryocar brasiliense) (EECP) por meio de avaliação morfológica (macroscópica, microscópica e ultramicroscópica), bem como avaliar a expressão de metaloproteinases (MMP2 e MMP9) e seus inibidores teciduais (TIMP1 e TIMP2) no miocárdio de ratos submetidos à cardiotoxicidade crônica pela DOX, tratados ou não com o EECP. O experimento teve duração de três meses e foram utilizados 30 ratos da raça Wistar, distribuídos em seis grupos de cinco animais. G1 e G2 receberam como pré-tratamento 300mg/kg e 600mg/kg de EECP, respectivamente, por gavagem, durante sete dias e mantiveram o tratamento durante os 21 dias de aplicação da DOX. Em G1, G2, G3, G4 e GC, a cardiotoxicidade foi induzida com aplicações semanais de 2mg/kg de DOX, via intraperitoneal, totalizando quatro aplicações (8mg/kg) e, nos ratos do grupo Sham (GS), foi aplicado 1ml de solução fisiológica. Os animais do G3 receberam diariamente 300mg/kg e os do G4 600mg/kg de EECP, por gavagem, durante os 21 dias de aplicação da DOX. Os do GC e GS receberam 1 ml de água, diariamente, também por gavagem. Após o término das aplicações, os animais foram mantidos por dois meses, totalizando três meses de experimento. A avaliação macroscópica foi realizada após 90 dias, momento em que foram colhidas amostras para análise em microscopia eletrônica, histopatologia e imunoistoquímica. Ao exame necroscópico foi observada ascite nos animais que receberam DOX. Houve baixo índice de mortalidade (3,33%), representado pela morte de um rato que desenvolveu pneumonia por falsa via. Não foi observada alteração no peso e nas medidas do coração dos ratos. Nas doses de 300 e 600mg/kg, o EECP atenuou a degeneração vacuolar miocítica. Na dose de 600mg/kg, o EECP reduziu a quantidade de células de Anitschkow e a fragmentação das miofibrilas. Não houve resultado significativo quanto à imunomarcação das MMP e, quanto a seus inibidores (TIMP), houve maior imunomarcação de TIMP2 no GC, grupo que recebeu apenas DOX. Concluiu-se que o extrato etanólico da casca do pequi (EECP) é eficiente em minimizar os efeitos da cardiotoxicidade crônica induzida pela DOX no miocárdio de ratos, considerando que nas doses de 300 e 600mg/kg o EECP atenua a degeneração vacuolar miocítica e, na dose de 600mg/kg, o EECP reduz a quantidade de células de Anitschkow e a fragmentação das miofibrilas.(AU)

Doxorubicin (DOX) is a chemotherapic drug used in the treatment of malignancies, but has the cardiotoxicity as collateral effect. The objective of this study was to evaluate the action of pequi shell etanolic extract (Caryocar brasiliense) (PSEE) through morphological evaluation (macroscopic, microscopic and ultramicroscopic), and to evaluate the expression of metalloproteinases (MMP2 and MMP9) and its tissue inhibitors (TIMP1 and TIMP2) in the myocardium of rats with chronic cardiotoxicity by DOX and treated or not with PSEE. The experiment lasted three months and 30 Wistar rats were divided into six groups of five animals. G1 and G2 received 300mg/kg and 600mg/kg of PSEE, respectively, as pretreatment, by gavage for seven days and continued treatment for 21 days of application of DOX. In G1, G2, G3, G4 and GC, cardiotoxicity was induced with weekly applications of 2mg/kg DOX, intraperitoneally, totaling four applications (8 mg/kg), and in the Sham group (GS) 1ml of saline solution was applied. G3 animals received daily 300mg/kg of PSEE, and G4, 600mg/kg, by gavage, for 21 days of application of DOX. The GC and GS received 1ml of water daily by gavage also. After the completion of the application, the animals were kept for two months, with three months of experiment. Macroscopic evaluation was performed after 90 days, at which time samples were taken for analysis in electron microscopy, histopathology and immunohistochemistry. At necropsy, ascites was observed in animals that received DOX. There was a low mortality rate (3.33%), being one mouse that developed false road pneumonia. There was no change in weights and measures of the rat hearts. At doses of 300 and 600mg/kg, the PSEE attenuates myocyte vacuolar degeneration. At a dose of 600mg/kg, PSEE reduces amount Anitschkow cells. There was no significant result on the immunostaining of MMP, but considering their inhibitors (TIMP) there was a greater immunostaining of TIMP2 in GC, the group that received only DOX. It was concluded that PSEE is effective in minimizing effects of chronic cardiotoxicity induced by DOX in the myocardium of rats, whereas at doses of 300 and 600mg/kg, PSEE attenuates vacuolar degeneration in myocytes and at the dose of 600mg/kg the PSEE reduces the amount of Anitschkow cells and myofibrils fragmentation.(AU)

Utilization of dynamic light scattering to evaluate Pterodon emarginatus oleoresin-based nanoemulsion formation by non-heating and solvent-free method

Abstract Pterodon emarginatus Vogel, Fabaceae, is a great source of bioactive compounds. The most known and studied herbal derivative from this species is an ambar-colored oleoresin that contains vouacapane diterpenes and volatile terpenoids, such as β-caryophyllene. Some recent papers aimed to generate nanoemulsions using this oleoresin for biological applications. However, they used high-energy methods that elevate costs of the process or heating procedures, which offer the disadvantage of possible volatile substances loss. Thus, as part of our ongoing studies with nanobiotechnology of natural products, especially regarding preparation of nanoemulsions with promising plant-based oils by low cost and low energy methods, we decided to evaluate the ability of non-heating and solvent-free method to generate P. emarginatus oleoresin-based nanoemulsions. Two non-ionic surfactants were used to generate the nanoemulsions by a simple homogenization method with vortex stirrer. Low mean droplet size (<180 nm) and low polydispersity index (<0.200) were observed even after one day of preparation. The low coefficient of variation for the analyzed parameters of different batches and similar profile for droplet size distribution suggested reproducibility of the method. After 30 days, some degree of droplet growth was observed on nanoemulsion prepared with polyethyleneglycol 400 monooleate, while almost no alteration was observed for nanoemulsion prepared with polysorbate 85. Programmed temperature ramp analysis revealed that no major effects on droplet size and polydispersity index were observed, suggesting the robustness of formed nanoemulsions. Thus, the present study shows for the first time the formation of sucupira-based nanoemulsions by a simple, low cost and ecofriendly method. This study opens new perspectives for bioactive evaluation of this novel nano-product.

Engineering spray-dried rosemary extracts with improved physicomechanical properties: a design of experiments issue

ABSTRACT A 33 Box–Behnken design and Response Surface Methodology were performed to evaluate the influence of extract feed rate, drying air inlet temperature and spray nozzle airflow rate on the process yield, stability parameters (moisture content and water activity) and on several physicomechanical properties of spray-dried rosemary extracts. Powder yield ranged from 17.1 to 74.96%. The spray-dried rosemary extracts showed moisture content and water activity below 5% and 0.5%, respectively, which indicate their chemical and microbiological stabilities. Even without using drying aids, some sets of experimental conditions rendered dried products with suitable flowability and compressibility characteristics for direct preparation of solid dosage forms. Analysis of variance and Response Surface Methodology proved that studied factors significantly affected most of the spray-dried rosemary extract quality indicators at different levels. The main processing parameter affecting the spray-dried rosemary extract characteristics was inlet temperature. The best combination of parameters used to obtain a reasonable yield of stable dry rosemary extracts with adequate technological properties for pharmaceutical purpose involves an extract feed rate of 2 ml/min, 80 °C inlet temperature and 40 l/min SA. The design of experiments approach is an interesting strategy for engineering spray-dried rosemary extracts with improved characteristics for pharmaceutical industrial purpose.

Influence of environmental factors on the composition of essential oils from leaves of Myrcia tomentosa (Aubl) DC / Influencia de los factores ambientales sobre la composición de los aceites esenciales de hojas de Myrcia tomentosa (Aubl) DC

The leaves of Myrcia tomentosa were collected from five sites and four different months in the Brazilian Cerrado and their essential oils were obtained by hidrodistillation and analyzed by GC/MS. The aim of this work was to evaluate the influence of environmental factors on essential oils composition in the leaves of M. tomentosa. The results were submitted to stepwise Multiple Regression and Canonical Correlation Analysis that indicated a relationship between compounds in essential oils and some environmental factors (foliar nutrients, soil nutrients and climatic data). Cluster Analysis indicated a high chemovariability in the essential oils from different sites, also showed that the collection time had a minor effect on oil composition.

Las hojas de Myrcia tomentosa se obtuvieron de cinco sitios diferentes y cuatro meses en el Cerrado brasileño y sus aceites esenciales fueron obtenidos por hidrodistillation y analizados por GC/MS. El objetivo de este trabajo fue evaluar la influencia de los factores ambientales sobre la composición de los aceites esenciales en las hojas de M. tomentosa. Los resultados fueron sometidos a múltiples etapas de regresión y análisis de correlación canónica indicó que la relación entre los compuestos en los aceites esenciales y algunos factores ambientales (nutrientes foliares, fertilizantes y datos climáticos). El análisis de agrupamiento indicó chemovariability un alto en los aceites esenciales de diferentes sitios, también mostró que el equipo de recolección tuvo un efecto menor sobre la composición del aceite.

Impact of different extraction methods on the quality of Dipteryx alata extracts

This study aimed to impact of different extraction methods on the quality of Dipteryx alata Vogel, Fabaceae, extracts from fruits. The major compounds found were the lipids 38.9% (w/w) and proteins 26.20% (w/w). The residual moisture was 7.20% (w/w), total fiber 14.50% (w/w), minerals 4.10% (w/w) and carbohydrate 9.10 % (w/w). The species studied has great potential in producing oil, but the content and type of fatty acids obtained is dependent on the method of extraction. The Blingh & Dyer method was more selective for unsaturated fatty acids and Shoxlet method was more selective for saturated fatty acids. The tannin extraction by ultrasound (33.70 % w/w) was 13.90% more efficient than extraction by decoction (29 % w/w).

Environmental factors affecting the concentration of phenolic compounds in Myrcia tomentosa leaves

Myrcia tomentosa (Aubl.) DC., Myrtaceae, found in Central Brazilian Cerrado and popularly known as "goiaba-brava", belongs to the Myrcia genus, which has several species with medicinal properties such as: hypoglycemic, diuretic, hypotensive, antidiarrheal, antimicrobial and antitumor. The present study aimed to analyzed the environmental influence on concentrations of phenolic metabolites in M. tomentosa leaves. Compounds assayed in the leaves were: total phenols, tannins by protein precipitation, hydrolysable tannins and total flavonoids and mineral nutrients, while soil fertility was also analyzed, all over during one year. The results were submitted to Pearson Correlation Analysis and stepwise Multiple Regression Analysis to investigate the relationship between phenolics and environment data. Analysis of variance and Cluster Analysis allowed indicated a high variability in samples from different sites. The results obtained suggests that content of phenolics from M. tomentosa leaves are influenced by environmental factors, particularly some foliar nutrients (N1, Ca1 and Mn1), soil nutrients (Ca s and Ks) and Rainfall.

Spray drying of Eugenia dysenterica extract: effects of in-process parameters on product quality

A 2³ full factorial design was used to assess the impact of spraying air flow rate (30-50 L/min), drying air inlet temperature (90-150 ºC) and extract feed rate (4-6 g/min) on the quality of Eugenia dysenterica DC., Myrtaceae, spray-dried extracts. Response surface methodology (RSM) was applied to analyze the significance of the effects of process factors on product quality and to obtain fitted equations to predict dry powder properties. Powder yields were satisfactory, ranging from 34.64 to 63.92%. The dried products showed moisture contents and water activities below 5% and 0.5, respectively. The recuperation ratios of total polyphenols, tannins and flavonoids ranged from 88.66 to 99.07%, 70.38 to 81.87% and 74.51 to 98.68%, respectively. Additionally, in some conditions the parameters related to dry product’s flowability and compressibility varied over a range acceptable for pharmaceutical purposes. RSM proved that studied factors significantly affected most of the quality indicators at different levels. The spray drying technology is an attractive and promising alternative for the development of intermediate phytopharmaceutical products of E. dysenterica.

Dissolution test of herbal medicines containing Passiflora sp.

The dissolution test is an essential tool to assess the quality of herbal medicines in the solid dosage form for oral use. This work aimed to evaluate the dissolution behavior of three herbal medicines in the form of capsules and tablet containing Passiflora, produced with powder or dried extract. Assay of total flavonoids and dissolution methods were validated and obtained results allowed the quantification of flavonoids with precision, accuracy and selectivity. The percentage of total flavonoids found was 2 percent for capsule A (containing only powder), 0.97 percent for capsule B (containing only dried extract) and 5.5 percent for tablet. Although the content was lower, the release of flavonoids present in the capsule containing dried extract was 12 percent higher over 30 min, with dissolved percentage values of 87 and 75, for the capsules containing extract and powder, respectively. The tablet containing dried extract presented dissolution of 76 percent, despite the higher content of flavonoids, which may be due to pharmacotechnical problems. Obtained data demonstrated the need to implement these tests in the quality control of herbal medicines, confirming the release of the active ingredients that underlie the pharmacological action of these medicines.