Daniele Rodrigues

Antioxidant activity of hydrophilic and lipophilic extracts of Brazilian blueberries.

Hydrophilic and lipophilic extracts of ten cultivars of Highbush and Rabbiteye Brazilian blueberries (Vaccinium corymbosum L. and Vacciniumashei Reade, respectively) that are used for commercial production were analysed for antioxidant activity by the FRAP, ORAC, ABTS and β-carotene-linoleate methods. Results were correlated to the amounts of carotenoids, total phenolics and anthocyanins. Brazilian blueberries had relatively high concentration of total phenolics (1,622-3,457 mg gallic acid equivalents per 100 g DW) and total anthocyanins (140-318 mg cyanidin-3-glucoside equivalents per 100 g DW), as well as being a good source of carotenoids. There was a higher positive correlation between the amounts of these compounds and the antioxidant activity of hydrophilic compared to lipophilic extracts. There were also significant differences in the level of bioactive compounds and antioxidant activities between different cultivars, production location and year of cultivation.

Carotenoid esters in foods - A review and practical directions on analysis and occurrence

Carotenoids are naturally found in both free form and esterified with fatty acids in most fruits and some vegetables; however, up to now the great majority of studies presents data on carotenoid composition only after saponification. The reasons for this approach are that a single xanthophyll can be esterified with several different fatty acids, generating a great number of different compounds with similar chemical and structural characteristics, thus, increasing the complexity of analysis compared to the respective saponified extract. This means that since UV/Vis spectrum does not change due to esterification, differentiation between free and acylated xanthophylls is dependent at least on elution order and mass spectrometry (MS) features. The presence of interfering compounds, especially triacylglycerides (TAGs), in the non-saponified extract of carotenoids can also impair carotenoid ester analyses by MS due to high background noise and ionization suppression since TAGs can be present in much higher concentrations than the carotenoid esters. This leads to the need of development of new and effective clean-up procedures to remove the potential interferents. In addition, only few standards of xanthophyll esters are commercially available, making identification and quantification of such compounds even more difficult. Xanthophyll esterification may also alter some properties of these compounds, including solubility, thermostability and bioavailability. Considering that commonly consumed foods are dietary sources of xanthophyll esters and that it is the actual form of ingestion of such compounds, an increasing interest on the native carotenoid composition of foods is observed nowadays. This review presents a compilation of the current available information about xanthophyll ester analyses and occurrence and a practical guide for extraction, pre-chromatographic procedures, separation and identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

An in vitro digestion method adapted for carotenoids and carotenoid esters: moving forward towards standardization

In vitro digestion methods are a useful approach to predict the bioaccessibility of food components and overcome some limitations or disadvantages associated with in vivo methodologies. Recently, the INFOGEST network published a static method of in vitro digestion with a proposal for assay standardization. The INFOGEST method is not specific for any food component; therefore, we aimed to adapt this method to assess the in vitro bioaccessibility of carotenoids and carotenoid esters in a model fruit (Byrsonima crassifolia). Two additional steps were coupled to the in vitro digestion procedure, centrifugation at 20 000g for the separation of the aqueous phase containing mixed micelles and exhaustive carotenoid extraction with an organic solvent. The effect of electrolytes, enzymes and bile acids on carotenoid micellarization and stability was also tested. The results were compared with those found with a simpler method that has already been used for carotenoid bioaccessibility analysis. These values were in the expected range for free carotenoids (5-29%), monoesters (9-26%) and diesters (4-28%). In general, the in vitro bioaccessibility of carotenoids assessed by the adapted INFOGEST method was significantly higher (p < 0.05) than those assessed by the simplest protocol, with or without the addition of simulated fluids. Although no trend was observed, differences in bioaccessibility values depended on the carotenoid form (free, monoester or diester), isomerization (Z/E) and the in vitro digestion protocol. To the best of our knowledge, it was the first time that a systematic identification of carotenoid esters by HPLC-DAD-MS/MS after in vitro digestion using the INFOGEST protocol was carried out.

Polyphenol extraction optimisation from Ceylon gooseberry (Dovyalis hebecarpa) pulp

Originally from Asia, Dovyalis hebecarpa is a dark purple/red exotic berry now also produced in Brazil. However, no reports were found in the literature about phenolic extraction or characterisation of this berry. In this study we evaluate the extraction optimisation of anthocyanins and total phenolics in D. hebecarpa berries aiming at the development of a simple and mild analytical technique. Multivariate analysis was used to optimise the extraction variables (ethanol:water:acetone solvent proportions, times, and acid concentrations) at different levels. Acetone/water (20/80 v/v) gave the highest anthocyanin extraction yield, but pure water and different proportions of acetone/water or acetone/ethanol/water (with >50% of water) were also effective. Neither acid concentration nor time had a significant effect on extraction efficiency allowing to fix the recommended parameters at the lowest values tested (0.35% formic acid v/v, and 17.6 min). Under optimised conditions, extraction efficiencies were increased by 31.5% and 11% for anthocyanin and total phenolics, respectively as compared to traditional methods that use more solvent and time. Thus, the optimised methodology increased yields being less hazardous and time consuming than traditional methods. Finally, freeze-dried D. hebecarpa showed high content of target phytochemicals (319 mg/100g and 1,421 mg/100g of total anthocyanin and total phenolic content, respectively).

Two-step cleanup procedure for the identification of carotenoid esters by liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry

Carotenoids are naturally found in both free form and esterified with fatty acids in most fruits; however, up to now the great majority of studies only evaluated their composition after saponification. This fact is easily explained by the difficult to analyze carotenoid esters. Preliminary studies showed that cleanup procedures in the extract are necessary for further analysis by LC-MS/MS since triacylglycerols (TAGs) impair the MS detection. Considering these facts, we developed a new cleanup procedure to remove TAGs and other lipids from carotenoid fruit extracts. This procedure is based on physical removal of solid lipids at low temperature followed by open column chromatography on MgO and diatomaceous earth. Before cleanup, four carotenoid diesters and two free xanthophylls were identified in murici (Byrsonyma crassifolia), corresponding to about 65% of the total chromatogram area. After carrying out the two-step cleanup procedure, 35 carotenoids were identified, being 14 monoesters, six free carotenoids and 15 carotenoid diesters. We can conclude that this two-step procedure was successfully applied to murici, an Amazonian fruit, which contains high amounts of lipids.

Antioxidant capacity and composition of pitanga seeds

Food industry generates a significant amount of seed wastes from the juice production, frozen pulps and jams. Considering that the characterization of wastes is the first step to determine their potential use, the aim of the present study was to determine the composition and the antioxidant capacity of seeds from pitanga fruits with different flesh colors (purple, red and orange). Chemical composition results revealed that pitanga seeds are a good source of insoluble dietary fiber, with low protein and fat levels, and no relevant differences were found among pitanga seeds from different flesh colors. Pitanga seed extracts had powerful antioxidant capacity that was partially correlated to their high phenolic content and showed some variation according to the pitanga flesh colors. Accordingly, it’s suggested that this low value waste of pitanga processing, could be used as a source of natural antioxidants and dietary fiber, for animal and/or human nutrition.

Comparison of Two Static in Vitro Digestion Methods for Screening the Bioaccessibility of Carotenoids in Fruits, Vegetables, and Animal Products

In vitro digestion methods are routinely used to assess the bioaccessibility of carotenoids and other dietary lipophilic compounds. Here, we compared the recovery of carotenoids and their efficiency of micellarization in digested fruits, vegetables, egg yolk, and salmon and also in mixed-vegetable salads with and without either egg yolk or salmon using the static INFOGEST method22 and the procedure of Failla et al.16 Carotenoid stability during the simulated digestion was ≥70%. The efficiencies of the partitioning of carotenoids into mixed micelles were similar when individual plant foods and salad meals were digested using the two static methods. Furthermore, the addition of cooked egg or salmon to vegetable salads increased the bioaccessibility of some carotenoids. Our findings showed that the two methods of in vitro digestion generated similar estimates of carotenoid retention and bioaccessibility for diverse foods.

Biochemical Characterization of Dovyalis hebecarpa Fruits: A Source of Anthocyanins with High Antioxidant Capacity

UNLABELLED Dovyalis hebecarpa, which is also known as the Ceylon gooseberry, is an attractive exotic purple berry that is produced in the southwest of Brazil with high yields. Current literature lacks information about the nutritional composition and antioxidant capacity of this fruit. As such, this work aimed to evaluate and compare the proximate composition, phytochemical content, and antioxidant capacity of D. hebecarpa over two seasons through the application of a range of different methods, including FRAP, ABTS, and ORAC assays. The proximate composition and biometric parameters were monitored for quality purposes. The results significantly changed between seasons in accordance with variations in rain incidence and average temperatures. The Ceylon gooseberry contained a similar or higher content of anthocyanins (ACY) than other berries commonly consumed in human diet. These ACY were identified at higher levels in the fruits skin (284 to 351 mg of cyanidin-3-glucoside equivalents/100 g of fresh weight [FW]) than in its pulp (49 to 69 mg/100 g of FW). The use of an ORAC assay identified higher antioxidant activity (145 μmol of Trolox equivalents [TE]/g in the skins and 179 μmol of TE/g in the pulp on a FW basis) than all other methods used to determine antioxidant activity. Thus, D. hebecarpa is a rich source of ACY that demonstrates high antioxidant activity against the peroxyl radicals formed in ORAC assay conditions. PRACTICAL APPLICATION Ceylon gooseberry fruit is a source of phytochemicals that could be used in the human diet providing defense against free radicals damage. Moreover, the skins of the fruit, which are not typically consumed, contained higher levels of polyphenols than the pulp. This indicates that the skins of the Ceylon gooseberry may represent a promising source of natural pigments and antioxidants for industrial applications.

Evaluation of the isoflavone and total phenolic contents of kefir-fermented soymilk storage and after the in vitro digestive system simulation.

This study aimed to evaluate the isoflavone and total phenolic contents in kefir-fermented soymilk storage and after the in vitro digestive system simulation (DSS). Soymilk was fermented with kefir culture (0.02UC/L) at 25°C for 15h and stored at 4°C for 4days. After the fermentation and storage, the isoflavone and total phenolic contents were quantified by high performance liquid chromatography and spectrophotometry, respectively. The cell viability of lactic acid bacteria and yeast was evaluated. Fermentation promoted an increase of approximately 3log CFU/g cycles of the microorganisms and the storage process did not alter the aglycone isoflavones and total phenolic contents. The content of aglycone isoflavones increased 2-fold, and the total phenolic content increased 9-fold. Therefore, kefir-fermented soymilk is a good source of aglycone isoflavones and phenolics, since the content of these substances was increased significantly after the in vitro digestive system simulation of the product.

Effect of drying methods on the phenolic content and antioxidant capacity of Brazilian winemaking byproducts and their stability over storage

Abstract This work aimed to study the antioxidant capacity by different methods, the total content of polyphenols and the stability over time of dried byproducts from Brazilian hybrids and Vitis vinifera varieties. Oven-dried at 50 °C and spray-dried samples were monitored for 90 days of storage. Under testing conditions, BRS Violeta grapes showed the greatest stability and initial high levels of total phenolics and anthocyanins remained almost unchanged until the end of storage period. The same behavior was observed in BRS Violeta freeze-dried skins, seeds and lees (8557, 9520 and 4261 mg GAE/100 g DM, respectively, and 829 and 257 mg mv-3-glc/100 g DM in skin and lees, respectively). In all methodologies tested, BRS Violeta also showed higher values for antioxidant capacity. These results suggest that dried winemaking byproducts can be used as rich sources of polyphenol compounds for industrial extractions with high stability and antioxidant capacity.