Bacterial Carotenoids: Extraction, Characterization, and Applications.

Natural carotenoids are secondary metabolites that exhibit antioxidant, anti-inflammatory, and anti-cancer properties. These types of compounds are highly demanded by pharmaceutical, cosmetic, nutraceutical, and food industries, leading to the search for new natural sources of carotenoids. In recent years, the production of carotenoids from bacteria has become of great interest for industrial applications. In addition to carotenoids with C40-skeletons, some bacteria have the ability to synthesize characteristic carotenoids with C30-skeletons. In this regard, a great variety of methodologies for the extraction and identification of bacterial carotenoids has been reported and this is the first review that condenses most of this information. To understand the diversity of carotenoids from bacteria, we present their biosynthetic origin in order to focus on the methodologies employed in their extraction and characterization. Special emphasis has been made on high-performance liquid chromatography-mass spectrometry (HPLC-MS) for the analysis and identification of bacterial carotenoids. We end up this review showing their potential commercial use. This review is proposed as a guide for the identification of these metabolites, which are frequently reported in new bacteria strains.

Phenolic Fingerprinting and Bioactivity Profiling of Extracts and Isolated Compounds from Gypothamnium pinifolium Phil.

Gypothamnium pinifolium Phil. (Asteraceae) is a small shrub that grows in the Paposo Valley of the II Antofagasta Region of Chile. This initial study is of the high-resolution phenolic fingerprinting, antioxidant activity, the relaxation effects in rat aorta, the inhibitory enzyme potential, plus the antiproliferative activity of the ethyl acetate and n-hexane extract from G. pinifolium and its two major isolated secondary metabolites (one coumarin: 2-nor-1,2-secolycoserone, and one diterpene: ent-labda-8,13-E-diene-15-ol). The study involves using ultra-high-performance liquid chromatography todiode array detection coupled with Q-Orbitrap mass spectrometry analysis (UHPLC-PDA-Orbi-trap-MS), in which various compounds were identified, including specific coumarins. The n-hexane extract showed total phenolic and flavonoid contents of 517.4 ± 12.5 mg GAE/100 g extract and 72.3 ± 3.7 mg QE/100 g extract, respectively. In addition, the antioxidant activity of the n-hexane extract was assessed using in-vitro assays such as bleaching of DPPH and ABTS (IC50: 14.3 ± 0.52 and 2.51 ± 0.43 µg extract/mL, respectively), FRAP (347.12 ± 1.15 µmol Trolox equivalent/g extract), and ORAC (287.3 ± 1.54 µmol Trolox equivalents/g extract). Furthermore, the inhibition against cholinesterases (acetylcholinesterase (AChE) 4.58 ± 0.04 µg/mL, butyrylcholinesterase (BChE) IC50: 23.44 ± 0.03 µg/mL) and tyrosinase (IC50: 9.25 ± 0.15 µg/mL) enzymes of the n-hexane extract, and main compounds (IC50: 1.21 ± 0.03 µg/mL, 11.23 ± 0.02 µg/mL, 3.23 ± 0.12 µg/mL, and 103.43 ± 16.86 µg/mL, correspondingly for the most active coumarin 1) were measured. The antiproliferative potential of the extracts and the two principal compounds against several solid human cancer cells was investigated. All of them showed good activity against cancer cells. Label-free live-cell imaging studies on HeLa cells exposed to the isolated coumarin and the diterpene enabled the observation of cell death and several apoptotic hallmarks. Our results indicate that G. pinifolium Phil. is a valuable source of secondary metabolites with potential activity against noncommunicable diseases.

Pressurized natural deep eutectic solvents: An alternative approach to agro-soy by-products.

Soybeans are mainly used for food and biodiesel production. It is estimated that soy crops worldwide will leave about 651 million metric tons of branches, leaves, pods, and roots on the ground post-harvesting in 2022/23. These by-products might serve as largely available and cheap source of high added-value metabolites, such as flavonoids, isoflavonoids, and other phenolic compounds. This work aimed to explore green approaches based on the use of pressurized and gas expanded-liquid extraction combined with natural deep eutectic solvents (NADESs) to achieve phenolic-rich extracts from soy by-products. The total phenolic and flavonoid contents of the generated extracts were quantified and compared with conventional solvents and techniques. Pressurized liquid extraction (PLE) with choline chloride/citric acid/water (1:1:11 - molar ratio) at 120°C, 100 bar, and 20 min, resulted in an optimized condition to generate phenolic and flavonoid-rich fractions of soy by-products. The individual parts of soy were extracted under these conditions, with their metabolic profile obtained by UHPLC-ESI-QToF-MS/MS and potential antioxidant properties by ROS scavenging capacity. Extracts of soy roots presented the highest antioxidant capacity (207.48 ± 40.23 mg AA/g), three times higher than soybean extracts (68.96 ± 12.30). Furthermore, Hansen solubility parameters (HSPs) were applied to select natural hydrophobic deep eutectic solvents (NaHDES) as substituents for n-heptane to defat soybeans. Extractions applying NaHDES candidates achieved a similar yield and chromatography profile (GC-QToF-MS) to n-heptane extracts.

Emerging Lipids from Arecaceae Palm Fruits in Brazil.

Arecaceae palm tree fruits (APTFs) with pulp or kernel rich in oil are widely distributed in six Brazilian biomes. APTFs represent a great potential for the sustainable exploitation of products with high added value, but few literature studies have reported their properties and industrial applications. The lack of information leads to underutilization, low consumption, commercialization, and processing of these fruit species. This review presents and discusses the occurrence of 13 APTFs and the composition, physicochemical properties, bioactive compounds, and potential applications of their 25 oils and fats. The reported studies showed that the species present different lipid profiles. Multivariate analysis based on principal component analysis (PCA) and hierarchical cluster analysis (HCA) indicated a correlation between the composition of pulp and kernel oils. Myristic, caprylic, capric, and lauric acids are the main saturated fatty acids, while oleic acid is the main unsaturated. Carotenoids and phenolic compounds are the main bioactive compounds in APTFs, contributing to their high oxidative stability. The APTFs oils have a potential for use as foods and ingredients in the cosmetic, pharmaceutical, and biofuel industries. However, more studies are still necessary to better understand and exploit these species.

Metabolite Profiling of Soy By-Products: A Comprehensive Approach.

Soy is the major oilseed crop as soybeans are widely used to produce biofuel, food, and feed. Other parts of the plant are left on the ground after harvest. The accumulation of such by-products on the soil can cause environmental problems. This work presents for the first time a comprehensive metabolite profiling of soy by-products collected directly from the ground just after mechanical harvesting. A two-liquid-phase extraction using n-heptane and EtOH-H2O 7:3 (v/v) provided extracts with complete characterization by gas chromatography and ultra-high-performance liquid chromatography both coupled to time-of-flight mass spectrometry. A total of 146 metabolites, including flavones, flavonols, isoflavonoids, fatty acids, steroids, mono-, sesqui-, di-, and triterpenoids, were tentatively identified in soy by-products and soybeans. These proved to be sources of a wide range of bioactive metabolites, thus suggesting that they could be valorized while reducing potential environmental damage in line with a circular economy model.

One-step sustainable extraction of Silymarin compounds of wild Algerian milk thistle (Silybum marianum) seeds using Gas Expanded Liquids.

This work reports the application of Gas Expanded Liquid (GXL) extraction to concentrate the flavonolignan fraction (silymarin) and taxifolin from Silybum marianum seeds, which have proven to be highly valuable health-promoting compounds. GXL using green solvents was used to isolate silymarin with the objective of replacing conventional methods. In one hand, the effect of different compositions of solvents, aqueous ethanol (20%, 50% or 80% (v/v)) at different CO2/liquid (25, 50 and 75%) ratios, on the GXL extraction was investigated. The obtained extracts have been chemically and functionally characterized by means of UHPLC-ESI-MS/MS (triple quadrupole) and in-vitro assays such as anti-inflammatory, anti-cholinergic and antioxidant. Results revealed that the operating conditions influenced the extraction yield, the total phenolic content and the presence of the target compounds. The best obtained yield was 55.97% using a ternary mixture of solvents composed of CO2:EtOH:H2O (25:60:15) at 40 °C and 9 MPa in 160 min. Furthermore, the results showed that obtained extracts had significant antioxidant and anti-inflammatory activities (with best IC50 value of 8.80 µg/mL and 28.52 µg/mL, respectively) but a moderate anti-cholinesterase activity (with best IC50 value of 125.09 µg/mL). Otherwise, the concentration of silymarin compounds in extract can go up to 59.6% using the present one-step extraction method without further purification, being silybinA+B the predominant identified compound, achieving value of 545.73 (mg silymarin/g of extract). The obtained results demonstrate the exceptional potential of GXL to extract high-added values molecules under sustainable conditions from different matrices.

Comparison of Four Oil Extraction Methods for Sinami Fruit (Oenocarpus mapora H. Karst): Evaluating Quality, Polyphenol Content and Antioxidant Activity.

The sinami palm (Oenocarpus mapora H. Karst) is a plant from the South American Amazonia that has great potential for industrial applications in the development of functional foods, nutraceuticals and cosmeceuticals. In this manuscript, the physicochemical properties, total polyphenol content and antioxidant activity of sinami oil that was obtained using four extraction systems, namely expeller press extraction (EPE), cold press extraction (CPE), ultrasound-assisted extraction (UAE) and supercritical fluid extraction (SFE), were studied and compared. The oxidative stability (OSI) was statistically non-significant in EPE and SFE. The chromatic properties (CIELab) were influenced by the extraction methods and SFE presented high values of L* and a lower content of plant pigments. Ultrasound-assisted extraction showed a higher content of polyphenols and higher antioxidant activity. Different analyses for the evaluation of the physicochemical properties, the content of total polyphenols and antioxidant activity were used to classify sinami oil according to chemometrics using principal component analysis (PCA). For example, the sinami oil that was obtained using each extraction method was in a different part of the plot. In summary, sinami oil is an excellent resource for plant pigments. Additionally, the information that was obtained on the quality parameters in this study provided a good foundation for further studies on the characterization of major and minor compounds.

The role of spray-drying atmosphere on fridericia chica (bonpl.) L.G. Lohmann standardized extract production for wound healing activity.

Fridericia chica (Bonpl.) L.G. Lohmann (synonym Arrabidaea chica Verlot) is widely used in Brazilian folk medicine. Considering overcoming pitfalls of scaling up production of plant extracts, herein the effects of N2 atmosphere for extract spray-drying process is reported. Samples were monitored by in vitro antioxidant activity and microbiological evaluation. The drying atmosphere influenced 3-deoxyanthocyanines content when using air as atomizing gas, decreasing carajurin (37.5%) content with concomitant increase in luteolin yield (24.1%). Both drying processes preserved the pharmacological activity. In the cell migration test with HaCaT cells, the extract dried under air flow (5 µg/mL) promoted wound closure by 78% (12 hours) whereas the extract dried using N2 flow promoted 49% (12 hours), with 98% closure (12 hours) for the positive control. The antimicrobial evaluation for Staphylococcus aureus did not differ within drying atmospheres, with MIC (minimum inhibitory concentration) at 0.39 mg/mL. Therefore, the drying process reported herein did not interfere with the biological activity's outcome.

Comparison of different extraction methods of Brazilian "pacová" (Renealmia petasites Gagnep.) oilseeds for the determination of lipid and terpene composition, antioxidant capacity, and inhibitory effect on neurodegenerative enzymes.

Pacová (Renealmia petasites Gagnep.) is a Brazilian native plant, usually cultivated in south regions of the country. Pacová was previously reported concerning their possible health benefits, mostly from folk medicine. However, only few works relates the health benefits with the composition of the fruit parts. In this context, this work aimed to bring, for the first time in literature, the chemical characterization in respect to lipid and terpene composition of R. petasites oilseed, performed by three different extraction methods (supercritical fluid extraction (SFE) with CO2, Soxhlet with petroleum ether (SOX), and maceration with hexane (MAC)). SFE was most selective for MUFAs, PUFAs, sesqui- and diterpenes. The main terpene identified in all extracts was 2-carene. The extracts presented poor AChE inhibition, and SOX presented potential inhibitory effect against lipoxygenase activity. Overall, R. petasites oilseed is a natural source of terpenes and their potential health benefits are highly encouraged to be investigated.

Metabolomics as a Tool to Study Underused Soy Parts: In Search of Bioactive Compounds.

The valorization of agri-food by-products is essential from both economic and sustainability perspectives. The large quantity of such materials causes problems for the environment; however, they can also generate new valuable ingredients and products which promote beneficial effects on human health. It is estimated that soybean production, the major oilseed crop worldwide, will leave about 597 million metric tons of branches, leaves, pods, and roots on the ground post-harvesting in 2020/21. An alternative for the use of soy-related by-products arises from the several bioactive compounds found in this plant. Metabolomics studies have already identified isoflavonoids, saponins, and organic and fatty acids, among other metabolites, in all soy organs. The present review aims to show the application of metabolomics for identifying high-added-value compounds in underused parts of the soy plant, listing the main bioactive metabolites identified up to now, as well as the factors affecting their production.

Selective Extraction of Piceatannol from Passiflora edulis by-Products: Application of HSPs Strategy and Inhibition of Neurodegenerative Enzymes.

Passiflora edulis by-products (PFBP) are a rich source of polyphenols, of which piceatannol has gained special attention recently. However, there are few studies involving environmentally safe methods for obtaining extracts rich in piceatannol. This work aimed to concentrate piceatannol from defatted PFBP (d-PFBP) by means of pressurized liquid extraction (PLE) and conventional extraction, using the bio-based solvents selected with the Hansen solubility parameters approach. The relative energy distance (Ra) between solvent and solute was: Benzyl Alcohol (BnOH) < Ethyl Acetate (EtOAc) < Ethanol (EtOH) < EtOH:H2O. Nonetheless, EtOH presented the best selectivity for piceatannol. Multi-cycle PLE at 110 °C was able to concentrate piceatannol 2.4 times more than conventional extraction. PLE exhibited a dependence on kinetic parameters and temperature, which could be associated with hydrogen bonding forces and the dielectric constant of the solvents. The acetylcholinesterase (AChE) and lipoxygenase (LOX) IC50 were 29.420 µg/mL and 27.682 µg/mL, respectively. The results reinforce the demand for processes to concentrate natural extracts from food by-products.

Compressed fluids and phytochemical profiling tools to obtain and characterize antiviral and anti-inflammatory compounds from natural sources.

Many natural compounds, found mainly in plants, are associated with the treatment of various diseases. The search for natural therapeutic agents includes compounds with antiviral and anti-inflammatory activities. Among the many steps involved in bioprospection, extraction is the first and most critical step for obtaining bioactive compounds. One of the main advantages of using compressed fluids extraction is the high quality of the final product obtained due to the use of green solvents, while the selectivity towards target compounds can be tuned by adjusting the process parameters, especially pressure, temperature and solvent characteristics. In this review, a discussion is provided on the power of compressed fluids, such as supercritical fluid extraction (SFE), pressurized liquid extraction (PLE) and subcritical water extraction (SWE) to obtain antiviral and anti-inflammatory compounds from natural sources. In addition, an adequate knowledge about the identity and quantity of the compounds present in the extract is essential to correlate biological activity with chemical composition. Phytochemical profiling tools used for identification and quantification of these bioactive natural compound are also discussed. It can be anticipated that after the current SARS-COV-2 pandemic, the search of new natural compounds with antiviral and anti-inflammatory activity will be a hot research topic, so, this review provides an overview on the technologies currently used that could help this research.

Preparative Separation of Procyanidins from Cocoa Polyphenolic Extract: Comparative Study of Different Fractionation Techniques.

To provide further insight into the antioxidant potential of procyanidins (PCs) from cocoa beans, PC extract was fractionated by several methodologies, including solid phase extraction, Sephadex LH-20 gel permeation, and preparative HPLC using C18 and diol stationary phases. All the isolated fractions were analyzed by UHPLC-QTOF-MS to determine their relative composition. According to our results, classical techniques allowed good separation of alkaloids, catechins, dimers, and trimers, but were inefficient for oligomeric PCs. Preparative C18-HPLC method allowed the attainment of high relative composition of fractions enriched with alkaloids, catechins, and PCs with degree of polymerization (DP) < 4. However, the best results were obtained by preparative diol-HPLC, providing a separation according to the increasing DP. According to the mass spectrometry fragmentation pattern, the nine isolated fractions (Fractions II-X) consisted of exclusively individual PCs and their corresponding isomers (same DP). In summary, an efficient, robust, and fast method using a preparative diol column for the isolation of PCs is proposed. Regarding DPPH• and ABTS•+ scavenging activity, it increases according to the DP; therefore, the highest activity was for cocoa extract > PCs > monomers. Thereby, cocoa procyanidins might be of interest to be used as alternative antioxidants.

Food-Safe Process for High Recovery of Flavonoids from Cocoa Beans: Antioxidant and HPLC-DAD-ESI-MS/MS Analysis.

Considering the increasing interest in the incorporation of natural antioxidants in enriched foods, this work aimed to establish a food-grade and suitable procedure for the recovery of polyphenols from cocoa beans avoiding the degreasing process. The results showed that ultrasound for 30 min with particle sample size < 0.18 mm changed the microstructure of the cell, thus increasing the diffusion pathway of polyphenols and avoiding the degreasing process. The effect of temperature, pH, and concentration of ethanol and solute on the extraction of polyphenols was evaluated. Through a 24 full factorial design, a maximum recovery of 122.34 ± 2.35 mg GAE /g, 88.87 ± 0.78 mg ECE /g, and 62.57 ± 3.37 mg ECE /g cocoa beans, for total concentration of polyphenols (TP), flavonoids (TF), and flavan-3-ols (TF3), respectively, was obtained. Based on mathematical models, the kinetics of the solid-liquid extraction process indicates a maximum equilibrium time of 45 min. Analysis by HPLC-DAD-ESI-MS/MS showed that our process allowed a high amount of methylxanthines (10.43 mg /g), catechins (7.92 mg /g), and procyanidins (34.0 mg /g) with a degree of polymerization >7, as well as high antioxidant activity determined by Oxygen Radical Absorbance Capacity (1149.85 ± 25.10 µMTrolox eq /g) and radical scavenging activity (DPPH•, 120.60 ± 0.50 µM Trolox eq /g). Overall, the recovery method made possible increases of 59.7% and 12.8% in cocoa polyphenols content and extraction yield, respectively. This study showed an effective, suitable and cost-effective process for the extraction of bioactive compounds from cocoa beans without degreasing.

Foodomics evaluation of the anti-proliferative potential of Passiflora mollissima seeds.

The anti-proliferative potential of Passiflora mollissima seeds, an underexplored agri-food waste, was investigated in this work by evaluating the molecular changes induced at transcript and metabolite expression levels on HT-29 human colon cancer cells. For this purpose, a pressurized-liquid extract from P. mollissima seeds obtained under optimized conditions was used for the treatment of HT-29 cells and a multi-omics strategy applied, integrating transcriptomics and metabolomics analysis, along with viability and cell cycle assays to study the molecular mechanisms that explain the anti-proliferative activity of this fruit by-product. After treatment for 48 and 72 h, the viability of HT-29 colon cancer cells was markedly affected, whereas minor effects were observed on normal human colon fibroblast cells. The bioactive extract was shown to arrest HT-29 cells in the S and G2/M phases of the cell cycle, which might be mediated by the inactivation of the FAT10 cancer signalling pathway among other genes identified as altered in the transcriptomic analysis. In addition, cellular redox homeostasis, as well as the polyamines pathway and methionine metabolism were found to be affected as suggested from the metabolomics data. Finally, the Foodomics integration enabled the identification of genes, such as MAD2L1, involved in the polyamine and glutathione metabolism, or the inactivation of the NUPR1 transcription factor, that might be related with the alteration of the intracellular ceramide levels in response to endoplasmic reticulum stress.

El malestar emocional como síntoma de estrés postraumático / Emotional distress as a symptom of post-traumatic stress

El trastorno por estrés postraumático (TEPT) tiene una importante prevalencia en la población, por lo que conocer los mecanismos psicológicos para la vulnerabilidad y el mantenimiento del trastorno es claramente relevante. Entre dichos mecanismos, se ha postulado el constructo de malestar emocional como transdiagnóstico a diversos trastornos mentales. El objetivo del presente estudio fue poner a prueba un modelo hipotético que analizaba la asociación entre tres variables pertenecientes a dicho constructo (evitación experiencial, tolerancia al malestar e intolerancia a la incertidumbre inhibitoria) y el TEPT. Participaron 328 estudiantes universitarios (74.7% mujeres y 25.3% hombres, con una media de edad de 21 años) que habían experimentado al menos un acontecimiento traumático de acuerdo con los criterios del DSM-5, quienes completaron una batería de cuestionarios. Se efectuó un análisis de ecuaciones estructurales cuyos resultados apoyaron parcialmente el modelo hipotético. La evitación experiencial mostró asociarse positiva y significativamente con la tolerancia al malestar, la intolerancia a la incertidumbre inhibitoria y los síntomas del TEPT. La tolerancia al malestar también se vinculó de forma positiva y significativa con dichos síntomas. Sin embargo, la intolerancia a la incertidumbre inhibitoria no mostró relación con esta sintomatología. En apoyo a la evidencia empírica precedente, los hallazgos de este estudio inciden en el papel de la evitación experiencial y la tolerancia al malestar como variables moderadoras de la sintomatología postraumática. Asimismo, apuntan hacia la necesidad de considerar estas variables en las intervenciones psicológicas para el tratamiento del TEPT

Post-traumatic stress disorder or PTSD has an important prevalence in the population, so knowing the psychological mechanisms of vulnerability and maintenance of this disorder is clearly relevant. Among those mechanisms, the emotional distress construct has been postulated as a trans-diagnostic tool related to different mental disorders. The objective of the present study was to test a hypothetical model for the analysis of the association between three variables of this construct (experiential avoidance, distress tolerance, and inhibitory uncertainty intolerance) and PTSD. The subjects of study amounted to 328 university students (74.7% women and 25.3% men, with a median age of 21) who had experienced at least one traumatic event according to the DSM-5 criteria. They were made to complete a set of questionnaires. A structural equation analysis was carried out, with results partially coincident with the hypothetical model. Exponential avoidance showed a positive and significant association with distress tolerance, inhibitory uncertainty intolerance and PTSD symptoms. Distress tolerance was also associated with these symptoms in a positive and significant way. However, inhibitory uncertainty intolerance was not associated with this symptomatology. Following the preexisting empirical evidence, the findings of this study have an impact on the role of experiential avoidance and distress tolerance, as variables that moderate post-traumatic symptomatology. Furthermore, the findings point to the need to consider these variables in psychological interventions for the treatment of PTSD

Insight of Stability of Procyanidins in Free and Liposomal Form under an in Vitro Digestion Model: Study of Bioaccessibility, Kinetic Release Profile, Degradation, and Antioxidant Activity.

Small unilamellar and multilayered liposomes loaded with polymeric (epi)catechins up to pentamers were produced. The bioaccessibility, kinetic release profile, and degradation under in vitro gastrointestinal conditions were monitored by UHPLC-DAD-QTOF-MS/MS. The results show that all of the procyanidins underwent depolymerization and epimerization into small molecular oligomers and mainly to (epi)catechin subunits. Moreover, all of the liposome formulations presented higher bioaccessibility and antioxidant activity in comparison to their respective counterparts in non-encapsulated form. Similar results were obtained with procyanidins from cocoa extract-loaded liposomes. Namely, the bioaccessibility of dimer, trimer, and tetramer fractions from cocoa-loaded liposomes were 4.5-, 2.1-, and 9.3-fold higher than those from the non-encapsulated cocoa extract. Overall, the procyanidin release profile was dependent on their chemical structure and physicochemical interaction with the lipid carrier. These results confirmed that liposomes are efficient carriers to stabilize and transport procyanidins with the aim of enhancing their bioaccessibility at a controlled release rate.

Recent applications of on-line supercritical fluid extraction coupled to advanced analytical techniques for compounds extraction and identification.

In a previous review (Sánchez-Camargo et al., J. Sep. Sci. 40 (2017) 213-227), we discussed the application of on-line supercritical fluid extraction coupled to chromatographic techniques. This review includes an update of the most recent publications (from January 2016 till June 2018) on this topic, which employs advanced analytical techniques for extracting and identifying valuable analytes. Supercritical fluid extraction has been widely recognized as a green sample preparation technique, because it is efficient, environmentally friendly, powerful, and faster, offering the possibility of direct coupling to analytical instrumental techniques. Among those techniques, supercritical fluid chromatography has experienced an innovative progression in the last 10 years, and the most recent applications of supercritical fluid extraction are coupled to this advanced analytical tool. The general principles, both methodological and instrumental of on-line supercritical fluid extraction coupled to supercritical fluid chromatography are described here. Besides, applications of the mentioned coupling for analysing biological fluids, food, soil, and botanical samples are also presented and discussed. Finally, a brief description about the very recent on-line coupling of supercritical fluid extraction to ion mobility spectrometry is presented, as well as concluding remarks about the importance of using these coupled techniques in the near future.

Design, Fabrication, Characterization, and In Vitro Digestion of Alkaloid-, Catechin-, and Cocoa Extract-Loaded Liposomes.

Liposomes containing theobromine, caffeine, catechin, epicatechin, and a cocoa extract were fabricated using microfluidization and sonication. A high encapsulation efficiency and good physicochemical stability were obtained by sonication (75% amplitude, 7 min). Liposomes produced at pH 5.0 had mean particle diameter ranging from 73.9 to 84.3 nm. The structural and physicochemical properties of the liposomes were characterized by transmission electron microscopy, confocal fluorescence microscopy, and antioxidant activity assays. The release profile was measured by ultra-high performance liquid chromatography coupled to diode array detection. The bioaccessibility of the bioactive compounds encapsulated in liposomes was determined after exposure to a simulated in vitro digestion model. Higher bioaccessibilities were measured for all catechins-loaded liposome formulations as compared to nonencapsulated counterparts. These results demonstrated that liposomes are capable of increasing the bioaccessibility of flavan-3-ols, which may be important for the development of nutraceutical-enriched functional foods.