Extract from Opuntia ficus-indica cladode delays the Aedes aegypti larval development by inducing an axenic midgut environment.

This study evaluated the effects of acute exposure of Aedes aegypti third instar (L3 ) larvae to the saline extract of Opuntia ficus-indica cladodes on the biological cycle and fertility of the emerging adults. For this, larvae were treated for 24 h with the extract at » LC50 (lethal concentration to kill 50% of larvae), ½ LC50 or LC50 ; the development and reproduction of the emerged adults were evaluated after a recovery period of 9 days. The resistance of proteins in the extract to hydrolysis by L3 digestive enzymes and histomorphological alterations in the larval midgut were also investigated. The extract contained lectin, flavonoids, cinnamic derivatives, terpenes, steroids, and reducing sugars. It showed a LC50 of 3.71% for 48 h. The data indicated mean survival times similar in control and extract treatments. It was observed development delay in extract-treated groups, with a lower number of adults than in control. However, the females that emerged laid similar number of eggs in control and treatments. Histological evaluation revealed absence of bacterial and fungal microorganisms in the food content in midguts from larvae treated with cladode extract. Electrophoresis revealed that three polypeptides in the extract resisted to hydrolysis by L3 digestive proteases for 90 min. The lectin activity was not altered even after 24-h incubation with the enzymes. In conclusion, the extract from O. ficus-indica can delay the development of Ae. aegypti larvae, which may be linked to induction of an axenic environment at larval midgut and permanence of lectin activity even after proteolysis.

Development of an analytical method for determination of polyphenols and total tannins from leaves of Syzygium cumini L. Skeels.

Syzygium cumini L. Skeels belongs to Myrtaceae family. This species has been recognized by its antidiabetic, anti-inflammatory, and antimicrobial activities. Despite ever-increasing scientific interest for this species there is no pharmacopeia method for characterization and standardization of S. cumini yet. So, toward this aim, the objective of this work was to develop an efficient analytical methodology able to determine polyphenols and tannins content from leaves hydroethanolic extract of S. cumini using Folin-Ciocalteu method by ultraviolet absorption spectrophotometry (UV-Vis). The analytical methodology was developed for the first time in the literature for leaves of this specie shown to be fast and low-cost with results expressed through tannic acid equivalent (TAE). Moreover, the methodology presented selectivity with maximum absorption at 706 nm wavelength, linearity with R2>0.99; limit of detection 0.275 µg TAE mL-1 and 0.102 µg TAE mL-1; limit of quantification 1.046 µg TAE mL-1 and 0.912 µg TAE mL-1 for total polyphenols and total tannins, respectively. Furthermore, the methodology was accurate with recover value greater than 98%, as well as exact, reproductive, and robust with coefficient of variation values less than 15% for both compounds. All the results are found within the fixed limits according to RDC 166/2017.

Anatomy, histochemistry and oxalic acid content of the leaflets of Averrhoa bilimbi and Averrhoa carambola

Abstract Averrhoa bilimbi L. and A. carambola L., Oxalidaceae, are the only two species of the genus Averrhoa L. Their leaves are widely used in folk medicine as an adjuvant in the treatment of diabetes. Some species may contain, for example, calcium oxalate crystals, which may lead to risk in its use when there is predisposition of individuals with reduced renal activity. Therefore, there are still few studies on the content of oxalic acid present in them, highlighting the importance of this investigation. The objective of this work was to conduct a comparative anatomical and histochemical study between the species and determining its content of oxalic acid. Semipermanent histological slides were prepared, following common plant anatomy procedures, for analysis of the leaflets in optical microscopy, polarization and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry. To determine the total, soluble and insoluble oxalate content was used titration with potassium permanganate. The anatomical characterization allowed identifying the characters useful in the differentiation of the species. The histochemistry revealed the location of the metabolites. Chemical microanalyses demonstrated that the crystals are of calcium oxalate. A. carambola presented the highest levels of total oxalate and soluble oxalate. The study assists in the identification and quality control of A. bilimbi and A. carambola and brings new data on its oxalic acid content, which are important, in view of the medicinal use of the species.

Chromatographic profiles of extractives from leaves of Eugenia uniflora

ABSTRACT Eugenia uniflora L., Myrtaceae, popularly known as "pitanga", is used in traditional medicine due the properties attributed to its chemical content, these being mainly hydrolysable tannins and flavonoids. This study provides a qualitative and quantitative evaluation of chemical profile from leaves of E. uniflora. The HPLC analysis was carried out on a C18 column (4.6 mm × 250 mm, 5 µm) by gradient elution with methanol and water (acidified with trifluoracetic acid); and silica gel Plates 60-F 254 with 10-12 µm and 5-6 µm particles, respectively for TLC and High HPTLC analysis. The chromatographic data obtained from HPLC, TLC and HPTLC presented bands and peaks related to flavonoids (myricitrin and derivatives) and tannins (gallic and ellagic acids), which were observed from different samples. The chromatographic similarities enabled the building of a typical fingerprint for the herbal material. The similarity analysis of the sample data by Pearson correlation showed R values >0.9 among peaks (HPLC) and bands (HPTLC). In addition, the analytical methodology developed by HPLC enabled the satisfactory quantification of marker substances [ellagic acid = 0.22% and 0.20% (m/m); gallic acid = 0.20% and 0.43%; myricitrin = 0.42 and 1.74% (m/m) in herbal drug and crude extract, respectively]. The procedure was also validated in accordance with the assays required by Brazilian legislation. Thus, the HPTLC and HPLC methods developed in this study provide helpful and simple tools for the quality evaluation both qualitatively and quantitatively of raw materials and extractives from leaves of E. uniflora.

Characterization of Leaf Extracts of Schinus terebinthifolius Raddi by GC-MS and Chemometric Analysis.

BACKGROUND: Schinus terebinthifolius Raddi belongs to Anacardiacea family and is widely known as "aroeira." This species originates from South America, and its extracts are used in folk medicine due to its therapeutic properties, which include antimicrobial, anti-inflammatory, and antipyretic effects. The complexity and variability of the chemical constitution of the herbal raw material establishes the quality of the respective herbal medicine products. OBJECTIVE: Thus, the purpose of this study was to investigate the variability of the volatile compounds from leaves of S. terebinthifolius. MATERIALS AND METHODS: The samples were collected from different states of the Northeast region of Brazil and analyzed with a gas chromatograph coupled to a mass spectrometer (GC-MS). The collected data were analyzed using multivariate data analysis. RESULTS: The samples' chromatograms, obtained by GC-MS, showed similar chemical profiles in a number of peaks, but some differences were observed in the intensity of these analytical markers. The chromatographic fingerprints obtained by GC-MS were suitable for discrimination of the samples; these results along with a statistical treatment (principal component analysis [PCA]) were used as a tool for comparative analysis between the different samples of S. terebinthifolius. CONCLUSION: The experimental data show that the PCA used in this study clustered the samples into groups with similar chemical profiles, which builds an appropriate approach to evaluate the similarity in the phytochemical pattern found in the different leaf samples. SUMMARY: The leave extracts of Schinus terebinthifolius were obtained by turbo-extractionThe extracts were partitioned with hexane and analyzed by GC-MSThe chromatographic data were analyzed using the principal component analysis (PCA)The PCA plots showed the main compounds (phellandrene, limonene, and carene), which were used to group the samples from a different geographical location in accordance to their chemical similarity. Abbreviations used: AL: Alagoas, BA: Bahia, CE: Ceará, CPETEC: Center for Weather Forecasting and Climate Studies, GC-MS: Gas chromatograph coupled to a mass spectrometer, MA: Maranhão, MVA: Multivariate data analysis, PB: Paraíba, PC1: Direction that describes the maximum variance of the original data, PC2: Maximum direction variance of the data in the subspace orthogonal to PC1, PCA: Principal component analysis, PE: Pernambuco, PI: Piauí, RN: Rio Grande do Norte, SE: Sergipe.

Effect of the Ethyl Acetate Fraction of Eugenia uniflora on Proteins Global Expression during Morphogenesis in Candida albicans.

Candida albicans is able to switch from yeast to hyphal growth and this is an essential step for tissue invasion and establishment of infection. Due to the limited drug arsenal used to treat fungal infections and the constant emergence of resistant strains, it is important to search for new therapeutic candidates. Therefore, this study aimed to investigate by proteomic analysis the role of a natural product (Eugenia uniflora) in impairing hypha formation in C. albicans. We also tested the potential action of E. uniflora to prevent and treat oral candidiasis induced in a murine model of oral infection and the ability of polymorphonuclear neutrophils to phagocytize C. albicans cells treated with the ethyl acetate fraction of the extract. We found that this fraction greatly reduced hypha formation after morphogenesis induction in the presence of serum. Besides, several proteins were differentially expressed in cells treated with the fraction. Surprisingly, the ethyl acetate fraction significantly reduced phagocytosis in C. albicans (Mean 120.36 ± 36.71 yeasts/100 PMNs vs. 44.68 ± 19.84 yeasts/100 PMNs). Oral candidiasis was attenuated when C. albicans cells were either pre-incubated in the presence of E. uniflora or when the fraction was applied to the surface of the oral cavity after infection. These results were consistent with the reduction in CFU counts (2.36 vs. 1.85 Log10 CFU/ml) and attenuation of tissue damage observed with histopathological analysis of animals belonging to treated group. We also observed shorter true hyphae by direct examination and histopathological analysis, when cells were treated with the referred natural product. The E. uniflora ethyl acetate fraction was non-toxic to human cells. E. uniflora may act on essential proteins mainly related to cellular structure, reducing the capacity of filamentation and attenuating infection in a murine model, without causing any toxic effect on human cells, suggesting that it may be a future therapeutic alternative for the treatment of Candida infections.

Protoflavanones from the Wood Stem of Salvertia convallariodora.

Phytochemical-analysis of the ethyl acetate extract of stem wood of Salvertia convallariodora A. St.-Hil. (Vochysiaceae), a Brazilian Cerrado species, led to the isolation and full characterization of three new non-aromatic B-ring flavanones (1-3) as well as the terpene mixture of sericic acid (4), 24-hydroxytormentic acid (5); 24-hydroxytormentic acid glucosyl ester (6), and sericoside (7), all identified for the first time from S. convallariodora. The structures of the new flavanones (1-3) were established from IR, LC-PDA-qTOF-MS, and NMR spectral data, including 2D NMR experiments.

Morpho-anatomical study of Ageratum conyzoides

ABSTRACT Ageratum conyzoides L., belonging to the family Asteraceae, is a tropical plant found in some regions of Africa, Asia and South America. This species is popularly known as billy goat weed, “mentrasto” and “catinga-de-bode” and has a large variety of secondary metabolites and biological activities mentioned in the literature. The objective of this work was to contribute the pharmacobotanical standardization of A. conyzoides. Cross-sections were obtained, by hand, for microscopic characterization of root, stem, petiole and leaf blade; to the leaf blade were still made paradermal and longitudinal sections, scanning electron microscopy analysis and maceration. The analysis showed that secretory structures ducts are evidenced only in the petiole and the leaf blade. The root has parenchymatous medullar region; stem, petiole and leaf blade exhibit striated cuticle. Non-glandular trichomes are present in stem, petiole and leaf blade, while capitate glandular trichomes are present only in the leaf blade and are restricted to the abaxial face. These anatomical features are useful for diagnosis of the species and provide support to their quality control.

Chromatographic and Spectrophotometric Analysis of Phenolic Compounds from Fruits of Libidibia ferrea Martius.

BACKGROUND: Libidibia ferrea (Mart. ex Tul.) L.P. Queiroz (Fabaceae) is a tree which is native to Brazil, widely known as "Jucá," where its herbal derivatives are used in folk medicine with several therapeutic properties. The constituents, which have already been described in the fruit, are mainly hydrolysable tannins (gallic acid [GA] and ellagic acid [EA]). OBJECTIVE: The aim of this study was to investigate the phenolic variability in the fruit of L. ferrea by ultraviolet/visible (UV/VIS) and chromatographic methods (high-performance liquid chromatography [HPLC]/high-performance thin layer chromatography [HPTLC]). MATERIALS AND METHODS: Several samples were collected from different regions of Brazil and the qualitative (fingerprints by HPTLC and HPLC) and quantitative analysis (UV/VIS and HPLC) of polyphenols were performed. RESULTS: The HPTLC and HPLC profiles allowed separation and identification of both major analytical markers: EA and GA. The chemical profiles were similar in a number of spots or peaks for the samples, but some differences could be observed in the intensity or area of the analytical markers for HPTLC or HPLC, respectively. Regarding the quantitative analysis, the polyphenolic content by UV/VIS ranged from 13.99 to 37.86 g% expressed as GA or from 10.75 to 29.09 g% expressed as EA. The contents of EA and GA by liquid chromatography-reversed phase (LC-RP) method ranged from 0.57 to 2.68 g% and from 0.54 to 3.23 g%, respectively. CONCLUSION: The chemical profiles obtained by HPTLC or HPLC, as well as the quantitative analysis by spectrophotometry or LC-RP method, were suitable for discrimination of each herbal sample and can be used as tools for the comparative analysis of the fruits from L. ferrea. SUMMARY: The polyphenols of fruits of Libidibia ferrea can be quantified by UV/VIS and HPLCThe HPLC method was able to detect the gallic and ellagic acids in several samples of fruits of Libidibia ferreaThe phenolic profiles of fruits from Libidibia ferrea by HPTLC and HPLC were reproductible. Abbreviations used: HPTLC: high performance thin layer chromatography, HPLC: high performance liquid chromatography, UV-Vis: spectrophotometry.

Reversed-phase-liquid chromatography method for separation and quantification of gallic acid from hydroalcoholic extracts of Qualea grandiflora and Qualea parviflora.

BACKGROUND: Qualea parviflora and Qualea grandiflora (Vochysiaceae), commonly known in Brazil as "pau-terra" and "pau-terrinha," respectively, have been widely used in the treatment of ulcer and gastritis. These therapeutic effects are attributed to various compounds present in the plants, including phenolic compounds such as gallic acid, due to their important antioxidant activity. OBJECTIVE: The aim of the present study was to validate a high performance liquid chromatography with diode array detection (HPLC-DAD) method for the quantitative determination of gallic acid in the stem bark of Q. parviflora and Q. grandiflora hydroalcoholic extracts. MATERIALS AND METHODS: The chromatography analysis was successfully achieved on a Dionex column, Acclaim(®) 120 (250 mm × 4.60 mm, 5 µm) with a gradient elution of water and methanol at a flow rate of 0.8 mL/min and ultraviolet detection at 280 nm. RESULTS: The validation data, including linearity, precision, specificity, accuracy and robustness of this method demonstrated good reliability and sensitivity. CONCLUSION: The method is able to quantify gallic acid in the stem bark of both species. What is more, the chromatographic peaks showed good resolution and there are also the advantages of easy sample preparation and a short time between each injection.

Development of standardized extractive solution from Lippia sidoides by factorial design and their redox active profile

AbstractThe aim of this study was to evaluate the influences of variables of preparation on total flavonoids content from extractive solution of Lippia sidoides Cham., Verbenaceae. Thus a 23 factorial design was used to study the importance of plant proportion, the extraction method and solvent on the extraction of flavonoid. The methodology of determination of chemicals in factorial design was validated according to the parameters required by Brazilian Health Agency. The extraction solution was selected through a full factorial design where the best conditions to achieve the highest content of flavonoids were: 7.5% (w/v) of plant with ethanol 50% (v/v) as solvent. The polyphenols content was determined by LC method and its relationship with the antioxidant and free radical scavenging activities was evaluated. The free radical scavenging activities and antioxidant potentials were determined for different concentrations using various in vitro models. Our results indicate that extracts exhibited a significant dose-dependent antioxidant effect as evaluated by TRAP/TAR assays. Besides, we observed an antioxidant activity against hydroxyl radicals and nitric oxide, and protection against lipid peroxidation in vitro. Our results suggest that the extract presents significant in vitro antioxidant potential indicating promising perspectives for its use as pharmaceutical/or food additive.

Validation of a spectrophotometric methodology for the quantification of polysaccharides from roots of Operculina macrocarpa (jalapa)

The roots from Operculina macrocarpa (L.) Urb., Convolvulaceae, are widely used in Brazilian traditional medicine as a laxative and purgative. The biological properties of this drug material have been attributed to its polysaccharides content. Thus, the aim of this study was to evaluate the polysaccharide content in drug material from O. macrocarpa by spectrophotometric quantitative analysis. The root was used as plant material and the botanical identification was performed by macro and microscopic analysis. The plant material was used to validate the spectrophotometric procedures at 490 nm for the quantification of the reaction product from drug polysaccharides and phenol-sulfuric acid solution. The analytical procedure was evaluated in order to comply with the necessary legal requirements by the determination of the following parameters: specificity, linearity, selectivity, precision, accuracy and robustness. This study provides with a simple and valid analytical procedure (linear, precise, accurate and reproducible), which can be satisfactorily used for quality control and standardization of herbal drug from O. macrocarpa.

Simaroubaceae family: botany, chemical composition and biological activities

The Simaroubaceae family includes 32 genera and more than 170 species of trees and brushes of pantropical distribution. The main distribution hot spots are located at tropical areas of America, extending to Africa, Madagascar and regions of Australia bathed by the Pacific. This family is characterized by the presence of quassinoids, secondary metabolites responsible of a wide spectrum of biological activities such as antitumor, antimalarial, antiviral, insecticide, feeding deterrent, amebicide, antiparasitic and herbicidal. Although the chemical and pharmacological potential of Simaroubaceae family as well as its participation in official compendia; such as British, German, French and Brazilian pharmacopoeias, and patent registration, many of its species have not been studied yet. In order to direct further investigation to approach detailed botanical, chemical and pharmacological aspects of the Simaroubaceae, the present work reviews the information regarding the main genera of the family up to 2013.

Total phytosterol content in drug materials and extracts from roots of Acanthospermum hispidum by UV-VIS spectrophotometry

Acanthospermum hispidum DC., Asteraceae, is widely used in folk medicine in Brazil to treat respiratory diseases; this biological property has been attributed to its phytosterol content. This study evaluated the spectrophotometric assay method to quantify the total phytosterol content in raw materials and extracts from roots of A. hispidum. The procedure was based on the quantification at 625 nm after the Liebermann-Burchard reaction. The method was evaluated for linearity, repeatability, intermediate precision, accuracy and robustness. The date indicated that the procedure is a valid analytical tool for materials and herbal derivatives from A. hispidum. .

Anatomy of leaf and stem of Erythrina velutina

Erythrina velutina Willd., Fabaceae, known as "mulungu", is a tree of tropical regions, as northeastern Brazil. Its bark is used in folk medicine as tranquilizer, sedative and insomnia. This study aimed to characterize the stem and leaf anatomy and to provide subsidies to quality control of the plant drug due to its wide use in folk medicine as well as its differentiation from other species with the same popular name. Samples were collected at Cuité, in Paraíba State, Brazil, fixed in FAA50, semipermanent slides were made, following usual procedures in plant anatomy. The stem shows a cylindrical contour, covered by a uniseriate epidermis covered by a thickened cuticle. It shows claviform glandular and branched trichomes with uniseriate stalk. Secretory cavities are into the phloem. The leaf epidermis has branched and glandular trichomes and anisocytic and paracytic stomata, on both sides, with predominance of branched trichomes and stomata on abaxial surface. Secretory cavities in stem and leaf, types of trichomes and stomata, its location and distribution constitute diagnostic characters for this specie. The structural characterization of the stem and leaf allows its distinction from other ones of this genus, ensuring safety for commercial pharmacological uses, allowing certification of the authenticity of raw material.

Anti-inflammatory and antinociceptive activities of Phyllanthus niruri spray-dried standardized extract

Phyllanthus niruri L., Euphorbiaceae, spray-dried standardized extract was studied for its anti-inflammatory and antinociceptive activities in adult albino rats and mice. The anti-inflammatory effect of spray-dried standardized extract was observed in carrageenan-induced paw edema and thioglycolate-induced leukocyte migration, while antinociceptive effects were observed using Randall & Selitto, tail flick, and hot plate tests. This study showed that intraperitoneal spray-dried standardized extract at 100, 200, 800, or 1600 mg/kg reduced the vascular response in the inflammatory process of paw edema induced by 1% carrageenan. Oral spray-dried standardized extract at 100 or 200 mg/kg inhibited leukocyte migration to the site of inflammation induced by 3% thioglycolate. In rats, at 100 and 200 mg/kg intraperitoneally, the extract exhibited a marked peripheral analgesic effect in a Randall & Selitto assay and showed significant central analgesic activity in a hot plate and tail flick assay. In conclusion, this study suggested that Phyllanthus niruri spray-dried standardized extract has potent inflammatory and antinociceptive activities and that these activities are not modified by standard drying process, making it feasible to use the dry extract standardized to obtain a phytotherapic preparation and thus validating its use for the treatment of pain and inflammation disorders.

Comparative evaluation of UV/VIS and HPLC analytical methodologies applied for quantification of flavonoids from leaves of Bauhinia forficata

Spectrophotometric determinations (direct dilution or after acid hydrolysis) of flavonoid content in leaves of Bauhinia forficata Link, Fabaceae, from two different regions from Brazil were compared with contents of kaempferitrin quantified by HPLC analysis. The absence of kaempferitrin peak in one sample denotes the risk of LC-method failure. On the other hand, the both spectrophotometric assays showed satisfactory performance and do not underwent influence from absence of kaempferitrin. Since several flavonoids are present in the herbal material, different content of flavonoids were observed for each analytical procedure. However, a strong positive association could be detected among methods (R²>0.99). Although a conversion factor should be adopted to compare procedures, the data showed that the spectrophotometric methods remain as an important tool for analysis of complex matrices such herbal drugs, notably when there aren't any pharmacological or chemical marker established for the species. Moreover, it's possible to suggest that the techniques studied in this work exhibit similar performance under the conditions employed. Nevertheless, before the adoption of kaempferitrin as an analytical marker by HPLC for Bauhinia species, the pharmacological knowledge should be better developed. In this cases, the use of spectrophotometric assay provide higher assurance of the reproducibility of the efficacy and safety.

Pharmacological screening and acute toxicity of bark roots of Guettarda platypoda

Due to its folk use, scientific reports and phytochemical screening, the purpose of this work was to study the phytochemical and the biological properties of the methanol extract and to evaluate the anti-inflammatory activity as well as determine the acute toxicity, antitumor and cytotoxic activity of the root barks of Guettarda platypoda DC., Rubiaceae. In this analysis the presence of flavonoids and therpenoids were identified. These data and the ones in the literature indicated it as a potential antioxidant and motivated the cytotoxic analysis related with three tumoral cell strains as well as to evaluate its antitumoral activity (sarcoma 180 and Ehrlich carcinoma) in female mice. Due to the presence of esteroids and the previous study of the ethanolic extract, its anti-inflammatory activity and toxicity were also evaluated. Absence or low toxicity in 2000 mg/kg doses was verified and the attention to their phytochemical and pharmacological properties is constantly increasing.

Development and evaluation of emulsions from Carapa guianensis (Andiroba) oil.

Carapa guianensis, a popular medicinal plant known as "Andiroba" in Brazil, has been used in traditional medicine as an insect repellent and anti-inflammatory product. Additionally, this seed oil has been reported in the literature as a repellent against Aedes aegypti. The aim of this work is to report on the emulsification of vegetable oils such as "Andiroba" oil by using a blend of nonionic surfactants (Span 80® and Tween 20®), using the critical hydrophilic-lipophilic balance (HLB) and pseudo-ternary diagram as tools to evaluate the system's stability. The emulsions were prepared by the inverse phase method. Several formulations were made according to a HLB spreadsheet design (from 4.3 to 16.7), and the products were stored at 25°C and 4°C. The emulsion stabilities were tested both long- and short-term, and the more stable one was used for the pseudo-ternary diagram study. The emulsions were successfully obtained by a couple of surfactants, and the HLB analysis showed that the required HLB of the oil was 16.7. To conclude, the pseudo-ternary diagram identified several characteristic regions such as emulsion, micro-emulsion, and separation of phases.

Development of superparamagnetic microparticles for biotechnological purposes.

Aqueous suspensions containing magnetic microparticles have been increasingly used in biosciences and biotechnology. This work describes an experimental procedure to produce superparamagnetic microparticles. The particles were prepared based on the coprecipitation of iron salts in alkaline medium. Afterwards, characterization was performed. Characterization data demonstrated that magnetite was the dominant phase in the analyzed sample, and 50% of them were in the size range of 0.5-5 microm. The results suggest that the experimental protocol provided a simple synthesis route to produce superparamagnetic microparticles. Such properties may be very useful for biotechnological purposes.