Umbu Fruit Peel as Source of Antioxidant, Antimicrobial and α-Amylase Inhibitor Compounds.

Herein, the extraction of bioactive compounds from umbu fruit peel was optimized using thermal-assisted solid-liquid extraction. In parallel, antioxidant, antimicrobial, and inhibitory effects against α-amylase of optimized extract were also evaluated. The combination of operational conditions including the temperature (32-74 °C), ethanol concentration (13-97%), and solid/liquid ratio (1:10-1:60; w/v) was employed using a rotational central composite design for optimization. The extracts were evaluated for total phenolic compounds (TPC), total flavonoid compounds (TFC) and antioxidant capacity by ABTS•+, DPPH• and FRAP assays. The bioactive profile of the optimized extract was obtained by ultra-performance liquid chromatography coupled to quadrupole/time-of-flight mass spectrometry in electrospray ionization in both negative and positive modes. The statistically evaluated results showed that the optimal operational conditions for the recovery of bioactive compounds from umbu fruit peel included 74 °C, 37% ethanol, and a solid-liquid ratio of 1:38. Under these conditions, the obtained values were 1985 mg GAE/100 g, 1364 mg RE/100 g, 122 µmol TE/g, 174 µmol/TE g and 468 µmol Fe2+/g for TPC, TFC, ABTS•+, DPPH•, and FRAP assays, respectively. In addition, the optimized extract was effective against Gram-positive and Gram-negative bacteria (MBC ranged from 0.060 to 0.24 mg GAE/mL), as well as it was effective to inhibit α-amylase (IC50 value of 0.076 mg GAE/mL). The optimized extract showed to be mainly constituted by phenolic acids and flavonoids.

Improvement in phloroglucinol compound extracted from different parts of Eugenia umbelliflora monitored by LC-UV and antimicrobial activity.

The phloroglucinol eugenial C, eugenial D and eugenial E are the main active compounds in Eugenia umbelliflora fruits. This study aims to evaluate the extraction conditions of E. umbelliflora, using ethanol as solvent, focusing on the phloroglucinol and antimicrobial activity. In order to optimize the extraction conditions, ethanol 50, 70 and 90 oGL was used as a solvent in the proportions of 1:20 (w/v) of drug:solvent ratio (D:S), stirring (330 rpm) at room temperature during 4 h, monitored by LC-UV and antimicrobial assay. The LC-UV method developed was linear over a concentration range of 3.4-68.0, 5.3-106.0 and 5.0-100.8 µg.mL-1 of eugenial C, eugenial D and eugenial E, having LOQ of 1.68, 1.33 and 0.8 µg.mL-1, respectively. The fruits showed the best herbal raw material and showed the highest phloroglucinol concentration and activity against S. aureus, when extracted with ethanol 90oGL, during 4 h, at 1:20 of D:S.

Solvent Mixture Optimization in the Extraction of Bioactive Compounds and Antioxidant Activities from Garlic (Allium sativum L.).

Garlic is a health promoter that has important bioactive compounds. The bioactive extraction is an important step in the analysis of constituents present in plant preparations. The purpose of this study is to optimize the extraction with the best proportion of solvents to obtain total phenolic compounds (TPC) and thiosulfinates (TS) from dried garlic powder, and evaluate the antioxidant activities of the optimized extracts. A statistical mixture simplex axial design was used to evaluate the effect of solvents (water, ethanol, and acetone), as well as mixtures of these solvents, after two ultrasound extraction cycles of 15 min. Results showed that solvent mixtures with a high portion of water and pure water were efficient for TPC and TS recovery through this extraction procedure. According to the regression model computed, the most significant solvent mixtures to obtain high TPC and TS recovery from dried garlic powder are, respectively, the binary mixture with 75% water and 25% acetone and pure water. These optimized extracts presented oxygen radical absorbance capacity. Pure water was better for total antioxidant capacity, and the binary mixture of water-acetone (75:25) was better for DPPH scavenging activity. These optimized extracts can be used for industrial and research applications.

The selective obtaining of amyrins from Amazonian Protium oleoresins / La obtención selectiva de amirinas a partir de oleorresinas de Protium amazónicas

SUMMARY The oleoresin produced by species of genus Protium sp. is rich in alpha and betaamyrins, two triterpenes with many pharmacogical activities. Considering the need to make the improved obtainment of these products feasible, this study sought to optimize techniques for the extraction and isolation of amyrins from resin. Two methods of extraction (maceration and sonication) with different solvents were compared to direct isolation from crude resin. The isolation of triterpenes was performed by chromatographic columns and the yields of extracts and fractions were analyzed by analysis of variance. The best extraction solvent for amyrins was hexane for both maceration and sonication methods (38.16±2.06% and 37.67±8.21%, respectively). There was no statistical difference between these methods and the direct method (32.05±2.40%). Additionally, the direct method is cheaper and more environmentally friendly. Thus, this study showed that it is possible to obtain a large quantity of amyrins by means of cheap, fast and ecological methods.

RESUMEN La oleorresina producida por especies del género Protium sp. es rica en amirinas alfa y beta, dos triterpenos con muchas actividades farmacológicas. Esta investigación buscó optimizar las técnicas de extracción y aislamiento de amirinas de la resina para hacer factible la obtención mejorada de esos productos. Se compararon dos métodos de extracción (maceración y sonicación) con diferentes solventes con aislamiento directo de la resina cruda. El aislamento de los triterpenos se realizó mediante columnas cromatográficas y los rendimientos de extractos y fracciones fueron hechos mediante análisis de varianza. El mejor solvente para la extracción de amirinas fue el hexano para ambos métodos de maceración y sonicación (38,16±2,06% y 37,67±8,21%, respectivamente). No hubo diferencia estadística entre estos métodos y el método directo (32,05±2,40%). Además, el método directo es más barato y ecológico. De este modo, esta investigación demostró que es posible obtener una gran cantidad de amirinas a través de métodos rápidos, baratos y ecológicos.

Extraction and purification of astaxanthin from shrimp shells and the effects of different treatments on its content

Abstract In this study, the extraction process and purification technology of astaxanthin from shrimp shells were established, and the effects of different treatments on the content of astaxanthin were studied. The determination results of astaxanthin in various shrimp/crab shells showed that the astaxanthin content in the Procambarus clarkia shell reached 239.96 µg/g. The effects of cool-ventilated, sun-dried and cooking conditions on the content of astaxanthin during the treatment of shrimp shells were investigated respectively and fresh shrimp shells as best extraction source was determined. The nine groups orthogonal test design with four factors and three levels (L9(3)4) was used to analyze the optimization of extraction process of astaxanthin from shrimp shells with ethanol as an environmentally friendly extraction solvent. The optimum experimental condition including the solid-liquid ratio (1:7), extraction time (20 min) and temperature (50 ºC) was proposed with the maximum extraction yield of astaxanthin. Next, silica gel column chromatography was used to purify the crude extraction of astaxanthin, and the purity of astaxanthin increased from 0.34% to 85.1% (about 250 times), which has great applications in the high value utilization of shrimp shells resources and the development of astaxanthin-related products.

Development of a Validated High-Performance Liquid Chromatography Method and Optimization of the Extraction of Lignans from Piper cubeba.

Piper cubeba L. f. is a food seasoning, which contains secondary metabolites displaying several biological properties, such as cytotoxic, anti-inflammatory, and antiparasitic activities. The lignans (+)-dihydroclusin, (-)-clusin, (-)-cubebin, (-)-yatein, and (-)-haplomyrfolin were isolated, with (-)-haplomyrfolin reported for the first time in P. cubeba seeds. Chromatographic standards were used to develop a reliable reverse-phase high-performance liquid chromatography analytical method according to the Agência Nacional de Vigilância Sanitária and International Conference on Harmonization guidelines to quantitate these lignans in both P. cubeba seeds and their extracts. The extraction of the lignans was also optimized, with the best conditions being ultrasound-assisted extraction, with 84% aqueous ethanol for 38 min in a single extraction. This procedure allows for the extraction of more than 80% of the total lignans, which is better in comparison to other techniques, such as maceration and Soxhlet extraction.

PARAFAC HPLC-DAD metabolomic fingerprint investigation of reference and crossed coffees.

In this study two cultivars of Coffea arabica L., Bourbon (reference) and IPR101 (crossing) were analyzed. The extracts were prepared according to a simplex centroid design with four components, ethanol, ethyl acetate, dichloromethane, and hexane. Multiway data were obtained by HPLC-DAD analysis of the fifteen different mixtures for each cultivar. The PARAFAC methodology was used to investigate the chromatographic fingerprint. For both cultivars, Factor 1 was able to discriminate mixtures containing ethyl acetate as solvent. Factor 2 indicated that mixtures in pure ethanol and binary mixtures containing ethanol were the most efficient in extracting chlorogenic acids and factor 3 identified methylxanthines through spectrophotometric profile in all mixtures. Higher concentrations were obtained by the ethanol, dichloromethane and hexane ternary mixture for the Bourbon cultivar and by the quaternary mixture of these solvents with ethyl acetate for the IPR101 cultivar. Trigonelline and cafestol were extracted in both cultivars. The reference coffee showed higher relative abundances of cafestol ester, chlorogenic acids and trigonelline whereas the crossed coffee showed higher levels of caffeine. To confirm these results, UPLC-MS was used as a complementary method to confirm the presence of the metabolites in these extracts. The three way PARAFAC strategy determines correlations of HPLC-DAD chromatographic and spectral data simultaneously with samples permitting a more unambiguous assignment of metabolic groups than can be obtained treating chromatographic and spectral data separately by two way methods. This can provide higher quality chromatographic fingerprints for food chemistry.

Multifunctions of Pleurotus sajor-caju (Fr.) Singer: A highly nutritious food and a source for bioactive compounds.

A study with Pleurotus sajor-caju was conducted to: evaluate the nutritional and chemical composition of the fruiting bodies; optimize the preparation of bioactive phenolic extracts; and characterize the optimized extract in terms of bioactive compounds and properties. P. sajor-caju revealed an equilibrated nutritional composition with the presence of hydrophilic (sugars and organic acids) and lipophilic (tocopherols and PUFA) compounds. p-Hydroxybenzoic, p-coumaric and cinnamic acids were identified in the extract obtained with ethanol (30g/l ratio) at 55°C for 85min. This extract showed antioxidant properties (mainly reducing power and lipid peroxidation inhibition), antibacterial activity against MRSA and MSSA and cytotoxicity against NCI-H460, MCF-7 and HeLa. Furthermore, as the extract showed capacity to inhibit NO production in Raw 264.7 macrophages, molecular docking studies were performed to provide insights into the anti-inflammatory mechanism of action, through COX-2 inhibition by the phenolic acids identified.

Black bean anthocyanin-rich extracts as food colorants: Physicochemical stability and antidiabetes potential.

Black beans contain anthocyanins that could be used as colorants in foods with associated health benefits. The objective was to optimize anthocyanins extraction from black bean coats and evaluate their physicochemical stability and antidiabetes potential. Optimal extraction conditions were 24% ethanol, 1:40 solid-to-liquid ratio and 29°C (P<0.0001). Three anthocyanins were identified by MS ions, delphinidin-3-O-glucoside (465.1m/z), petunidin-3-O-glucoside (479.1m/z) and malvidin-3-O-glucoside (493.1m/z). A total of 32mg of anthocyanins were quantified per gram of dry extract. Bean anthocyanins were stable at pH 2.5 and low-temperature 4°C (89.6%), with an extrapolated half-life of 277days. Anthocyanin-rich extracts inhibited α-glucosidase (37.8%), α-amylase (35.6%), dipeptidyl peptidase-IV (34.4%), reactive oxygen species (81.6%), and decreased glucose uptake. Black bean coats are a good source of anthocyanins and other phenolics with the potential to be used as natural-source food colorants with exceptional antidiabetes potential.

Experimental designs characterizing seasonal variations and solvent effects on the quantities of coumarin and related metabolites from Mikania laevigata.

Statistical design mixtures of acetone, chloroform, dichloromethane and ethanol were used to study the effects of different solvents and their mixtures on the quantities of coumarin and related metabolites extracted from Mikania laevigata samples harvested in each of the four seasons. RP-HPLC-DAD and both positive and negative modes of UPLC-MS analyses were used to determine relative quantities of coumarin, o-coumaric acid and melilotic acids in each season for all the mixture design extracts. The existence and measurement of the relative abundances of melilotic acid in Mikania laevigata have not been reported previously. Highest coumarin concentrations were encountered in the summer whereas its o-coumaric acid precursor and melilotic acid were most abundant in the spring. O-coumaric and melilotic acids concentrations were strongly correlated during the year. Also solvent effects were seen to be significant. Ethanol and 1:1 binary mixtures of ethanol and acetone extracted the largest quantities of coumarin whereas ethanolic binary and ternary mixtures with chloroform and dichloromethane provided the best yields of o-coumaric and melilotic acids. Statistical mixture models indicated that synergic binary interactions, especially those involving ethanol with acetone or chloroform, are important in the Mikania extraction process.

Optimization of the extraction of curcumin from Curcuma longa rhizomes

The aim of this work was to study the effect of dynamic maceration factors upon the curcumin content of Curcuma longa L., Zingiberaceae, extracts and to determine the optimum set of parameters for the extraction of curcumin using a 2(5) full factorial design and the response surface methodology. Under the established conditions, the content of soluble solids and curcumin in the extracts ranged from 0.8 to 3.4%, and from 0.1 to 1.8%, respectively. The most influential variable observed for the extraction was the ethanolic strength of the solvent. The optimized condition involves an extraction time of 12 h, agitation speed of 30 rpm, drug to solvent ratio of 1/6, extraction temperature of 80 ºC and the solvent with ethanolic strength of 70%. The data reported herein are useful for further developments of curcuma phytopharmaceutical intermediate products with optimized characteristics.