Using High-Pressure Technology to Develop Antioxidant-Rich Extracts from Bravo de Esmolfe Apple Residues.

Bravo de Esmolfe (BE) is a traditional Portuguese apple highly appreciated by consumers due to its peculiar flavor and aroma. This apple contains higher concentration of phenolic compounds than other cultivars and is thus considered a rich source of antioxidants. Its sensorial and functional properties have attracted farmers' associations to increase BE production. However, a large quantity of apples is wasted due to storage/transportation procedures that impact BE's quality attributes. In this work, we applied high-pressure extraction methodologies to generate antioxidant-rich fractions from BE residues aiming at adding high value to these agro-food by-products. We performed a first extraction step using supercritical CO2, followed by a second extraction step where different CO2 + ethanol mixtures (10-100% v/v) were tested. All experiments were carried out at 25 MPa and 50 °C. Extracts were characterized in terms of global yield, phenolic content and antioxidant activity using chemical (ORAC, HOSC, HORAC) and cell-based assays (CAA). We demonstrated that, although the pressurized 100% ethanol condition promoted the highest recovery of phenolic compounds (509 ± 8 mg GAE/100 g BE residues), the extract obtained with 40% ethanol presented the highest CAA (1.50 ± 0.24 µmol QE/g dw) and ORAC (285 ± 16 µmol TEAC/g dw), as well as HOSC and HORAC values, which correlated with its content of epicatechin and procyanidin B2. Noteworthy, this fraction inhibited free radical production in human neurospheroids derived from NT2 cells, a robust 3D cell model for neuroprotective testing.

Data sharing in PredRet for accurate prediction of retention time: Application to plant food bioactive compounds.

Prediction of retention times (RTs) is increasingly considered in untargeted metabolomics to complement MS/MS matching for annotation of unidentified peaks. We tested the performance of PredRet (http://predret.org/) to predict RTs for plant food bioactive metabolites in a data sharing initiative containing entry sets of 29-103 compounds (totalling 467 compounds, >30 families) across 24 chromatographic systems (CSs). Between 27 and 667 predictions were obtained with a median prediction error of 0.03-0.76 min and interval width of 0.33-8.78 min. An external validation test of eight CSs showed high prediction accuracy. RT prediction was dependent on shape and type of LC gradient, and number of commonly measured compounds. Our study highlights PredRet's accuracy and ability to transpose RT data acquired from one CS to another CS. We recommend extensive RT data sharing in PredRet by the community interested in plant food bioactive metabolites to achieve a powerful community-driven open-access tool for metabolomics annotation.

Pyromellitic dianhydride crosslinked soluble cyclodextrin polymers: Synthesis, lopinavir release from sub-micron sized particles and anti-HIV-1 activity.

We report the synthesis of water soluble cyclodextrin (CD) polymers prepared by crosslinking pyromellitic dianhydride (PMDA) with two CD derivatives (methyl-ß-CD - MßCD and (2-hydroxy)propyl-ß-CD - HPßCD) and their evaluation as functional sub-micron sized carriers in the development of antiretroviral drug delivery systems. Using the protease inhibitor lopinavir (LPV) as model drug, LPV loaded CD polymers (pHPßCD and pMßCD) were prepared and fully characterized. The physicochemical characterization and in vitro drug release confirmed the successful synthesis of pHPßCD and pMßCD, the formation of sub-micron sized particles and a 12-14 fold increase in LPV solubility. Cytotoxicity assays indicated that both pHPßCD and pMßCD were able to improve the safety profile of LPV while the viral infectivity assay revealed a concentration independent anti-HIV-1 effect for both pHPßCD and pMßCD with a maximum percentage inhibition (MPI) of 79 and 91% respectively. After LPV loading, the antiviral profile of pHPßCD was reversed to the sigmoidal dose-response profile of LPV, while pMßCD maintained its dose-independent profile followed by a LPV mediated increase in viral inhibition. Overall, both pHPßCD and pMßCD demonstrated anti-HIV-1 activity, while drug loaded pMßCD indicated its potential as functional sub-micron sized drug delivery polymers for achieving synergistic anti-HIV activity.

Factors Explaining Interpersonal Variation in Plasma Enterolactone Concentrations in Humans.

Lignans are diphenolic plant compounds with potential health modulating properties that are absorbed to the circulation and metabolized to the enterolignans enterodiol (END) and enterolactone (ENL) by gut microbiota. Epidemiological studies have inconsistently shown that a high lignan intake and circulating ENL are associated with reduced risk of breast-, prostate-, and colorectal cancer as well as cardiovascular disease and total and cause-specific mortality. Inconsistencies can be due to interpersonal variation of ENL formation or responses. The aim of this review is to identify and evaluate the impact of factors influencing variability in plasma concentrations of the main enterolignan, ENL. The main determinants of plasma ENL concentrations are intake of lignan and lignan-rich foods, composition and activity of intestinal microflora, antimicrobial use, nutrient intake, BMI, smoking, sex, and age. Composition and activity of the intestinal microbiota appear to be the most critical factor governing interpersonal variability in plasma ENL concentration followed by the use of antibiotics. Future studies with combined data from gut microbiota and metabolomics with food intake and life style data can be used to estimate the relative contribution of the different factors to ENL concentration in quantitative terms.

Interlaboratory Coverage Test on Plant Food Bioactive Compounds and their Metabolites by Mass Spectrometry-Based Untargeted Metabolomics.

Bioactive compounds present in plant-based foods, and their metabolites derived from gut microbiota and endogenous metabolism, represent thousands of chemical structures of potential interest for human nutrition and health. State-of-the-art analytical methodologies, including untargeted metabolomics based on high-resolution mass spectrometry, are required for the profiling of these compounds in complex matrices, including plant food materials and biofluids. The aim of this project was to compare the analytical coverage of untargeted metabolomics methods independently developed and employed in various European platforms. In total, 56 chemical standards representing the most common classes of bioactive compounds spread over a wide chemical space were selected and analyzed by the participating platforms (n = 13) using their preferred untargeted method. The results were used to define analytical criteria for a successful analysis of plant food bioactives. Furthermore, they will serve as a basis for an optimized consensus method.

Characterization by liquid chromatography-mass spectrometry and antioxidant activity of an ethanolic extract of Inula viscosa leaves.

Inula viscosa (I. viscosa) is a common Mediterranean plant, well known for its content on bioactive molecules. The chemical composition of an ethanolic extract from I. viscosa leaves, growing in Algeria, was analysed by liquid chromatography coupled to photodiode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI-MS/MS) operating in negative and positive mode. The methodology used revealed the presence of 51 compounds from which 47 were putatively identified, including 11 phenolic acids, 23flavonoids, one lignan and 12 terpenoids. Twenty-six of these compounds are described for the first time in I. viscosa. Antioxidant activity was measured using three different and complementary chemical assays: DPPH radical scavenging activity, oxygen radical absorbance capacity (ORAC) and hydroxyl radical scavenging capacity (HOSC). Results demonstrate that ethanolic leaf extract exhibit a high scavenging ability against DPPH (157.72 ±â€¯6.45 µM TE/g DW), peroxyl (4471.42 ±â€¯113.16 µM TE/g DW) and hydroxyl (630.10 ±â€¯17.81 µM TE/g DW) radicals, indicating that I. viscosa can be a promising source of bioactive compounds.

Polymethoxylated Flavones from Orange Peels Inhibit Cell Proliferation in a 3D Cell Model of Human Colorectal Cancer.

Polymethoxylated flavones (PMFs) have been recognized to inhibit colorectal cancer proliferation through various mechanisms, however most of these studies have been performed on cells grown as monolayers that present limitations in mimicking the 3D tumor architecture and microenvironment. The main aim of this study was to investigate the anticancer potential of an orange peel extract (OPE) enriched in PMFs in a 3D cell model of colorectal cancer. The OPE was developed by supercritical fluid extraction and the anticancer effect was evaluated in HT29 spheroids cultures in a stirred-tank based system. Results showed that OPE inhibited cell proliferation, induced cell cycle arrest (G2/M phase), promoted apoptosis, and reduced ALDH+ population on HT29 spheroids. The antiproliferative activity was significantly lower than that obtained for 2D model (EC50 value of 0.43 ± 0.02 mg/mL) and this effect was dependent on diameter and cell composition/phenotype of spheroids derived from different culture days (day 3 - 0.53 ± 0.05 mg/mL; day 5 - 0.55 ± 0.03 mg/mL; day 7 - 1.24 ± 0.15 mg/mL). HT29 spheroids collected at day 7 presented typical characteristics of in vivo solid tumors including a necrotic/apoptotic core, hypoxia regions, presence of cancer stem cells, and a less differentiated invasive front. Nobiletin, sinesentin, and tangeretin were identified as the main compounds responsible for the anticancer activity.

Characterization of phenolic compounds in chia (Salvia hispanica L.) seeds, fiber flour and oil.

The consumption of chia seeds products has increased recently and it has been suggested that the inclusion of this functional food in a daily human diet could contribute to improve consumers' health. However, a better knowledge about the composition of these products is mandatory. In this work, the phenolic compounds from commercial samples of chia seed, fiber flour and oil were extracted using an ultrasound-assisted methodology and were separated and identified by high-performance liquid chromatography coupled to a mass spectrometer. Methanol:water extracts were prepared and submitted to an acidic hydrolysis. Crude and hydrolyzed extracts were analyzed and phenolic compounds found were mainly caffeic acid and danshensu and its derivatives, such as rosmarinic and salvianolic acids. TPC was higher in the hydrolyzed extracts. These results supply new information about the main phenolic compounds presents in chia, which are important dietary sources of natural antioxidants for prevention of diseases caused by oxidative stress.