In Vitro Screening and Lipid-Lowering Effect of Prickly Pear (Opuntia Ficus-Indica L. Mill.) Fruit Extracts in 3T3-L1 Pre-Adipocytes and Mature Adipocytes.

Opuntia ficus-indica fruits have been widely used due to their nutritional composition and beneficial effects on health, particularly against chronic diseases such as diabetes, obesity, cardiovascular diseases and cancer, among others. In recent years, prickly pear peel and pulp extracts have been characterised, and a high number of bioactive compounds have been identified. This study aimed to analyse the triglyceride-lowering effect of prickly pear peel and pulp extracts obtained from fruits of three varieties (Pelota, Sanguinos, and Colorada) in 3T3-L1 maturing and mature adipocytes. At a concentration of 50 µg/mL, peel extracts from Colorada reduced triglyceride accumulation in pre-adipocytes and mature adipocytes. Additionally, at 25 µg/mL, Pelota peel extract decreased triglyceride content in mature adipocytes. Moreover, maturing pre-adipocytes treated with 50 and 25 µg/mL of Sanguinos pulp extract showed a reduction of triglyceride accumulation. In addition, the lipid-lowering effect of the main individual betalain and phenolic compounds standards were assayed. Piscidic acid and isorhamnetin glycoside (IG2), found in Colorada peel extract, were identified as the bioactive compounds that could contribute more notably to the triglyceride-lowering effect of the extract. Thus, the betalain and phenolic-rich extracts from Opuntia ficus indica fruits may serve as an effective tool in obesity management.

Brazil Nut (Bertholletia excelsa) Beverage Processed by High-Pressure Homogenization: Changes in Main Components and Antioxidant Capacity during Cold Storage.

High-pressure homogenization (HPH) is an emerging technology for obtaining physical and microbial stability of plant-based milks, but there is little information on the effects of this technology on the phytochemical components of the processed plant food beverage and during its cold storage. The effect of three selected HPH treatments (180 MPa/25 °C, 150 MPa/55 °C, and 50 MPa/75 °C) and pasteurization (PAS) (63 °C, 20 min) on minor lipid constituents, total proteins, phenolic compounds, antioxidant capacity, and essential minerals of Brazil nut beverage (BNB) were studied. Additionally, the study of the possible changes in these constituents was carried out during cold storage at 5 °C for 21 days. The fatty acid profile (dominated by oleic acid and linoleic acid), free fatty acid content, protein, and essential minerals (notable source of Se and Cu) of the processed BNB remained almost stable to treatments (HPH and PAS). Specifically, reductions in squalene (22.7 to 26.4%) and γ-γ-tocopherol (28.4 to 36%) were observed in beverages processed via both non-thermal HPH and thermal PAS, but ß-sitosterol remained unchanged. Total phenolics were reduced (24 to 30%) after both treatments, a factor that influenced the observed antioxidant capacity. The studied individual phenolics in BNB were gallic acid, catechin, epicatechin, catechin gallate, and ellagic acid, being the most abundant compounds. During cold storage (5 °C) up to 21 days, changes in the content of phytochemicals, minerals, and total proteins were not noticeable for any treated beverages, and no lipolysis processes were promoted. Therefore, after the application of HPH processing, Brazil nut beverage (BNB) maintained almost unaltered levels of bioactive compounds, essential minerals, total protein, and oxidative stability, remarkable characteristics for its potential development as a functional food.

A comprehensive review on carotenoids in foods and feeds: status quo, applications, patents, and research needs.

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted.

Anti-Obesity Effects of Isorhamnetin and Isorhamnetin Conjugates.

Isorhamnetin is a plant-derived secondary metabolite which belongs to the family of flavonoids. This review summarises the main outcomes described in the literature to date, regarding the effects of isorhamnetin on obesity from in vitro and in vivo studies. The studies carried out in pre-adipocytes show that isorhamnetin is able to reduce adipogenesis at 10 µM or higher doses and that these effects are mediated by Pparγ and by Wnt signalling pathway. Very few studies addressed in rodents are available so far. It seems that treatment periods longer than two weeks are needed by isorhamnetin and its glycosides to be effective as anti-obesity agents. Nevertheless, improvements in glycaemic control can be observed even in short treatments. Regarding the underlying mechanisms of action, although some contradictory results have been found, reductions in de novo lipogenesis and fatty acid uptake could be proposed. Further research is needed to increase the scientific evidence referring to this topic; studies in animal models are essential, as well as randomised clinical trials to determine whether the positive results observed in animals could also be found in humans, in order to determine if isorhamnetin and its glycosides can represent a real tool against obesity.

Effect of Ultrasound-Assisted Extraction of Carotenoids from Papaya (Carica papaya L. cv. Sweet Mary) Using Vegetable Oils.

By-products from fruits and are of great interest for their potential use in the food industry due to their high content of bioactive compounds. Herein, we examined the ultrasound-assisted extraction (UAE) of carotenoid and carotenoid esters from papaya pulp and peel using soybean oil and sunflower oil as alternative green solvents. Response surface methodology (RSM) was established to optimize the UAE process. Three independent variables, ultrasonic amplitude (20-60%), time (10-60 min), and co-solvent percentage (ethanol) (5-20%, v/v), were applied. The highest total carotenoid content in the UAE extracts was obtained from papaya pulp extracts (58.7 ± 1.6 and 56.0 ± 1.5 µg carotenoids/g oil) using soybean oil and sunflower oil, respectively (60% amplitude/ 10 min/ 20% ethanol). On the other hand, the highest carotenoid content (52.0 ± 0.9 µg carotenoids/g oil) was obtained from papaya peel using soybean oil applying the UAE process (20% amplitude/ 77 min/ 20% ethanol); a minor content of 39.3 ± 0.5 µg carotenoids/g oil was obtained from papaya peel using sunflower oil at 60% amplitude/ 60 min/ 5% ethanol. Lycopene was the most abundant carotenoid among all individual carotenoids observed in papaya oil extracts, obtaining the highest yields of this carotenoid when papaya pulp and peel were extracted using soybean oil (94% and 81%, respectively) and sunflower oil (95% and 82%, respectively). Great extraction of xanthophyll esters was detected using 20% of ethanol in the vegetable oil extraction solvent (v/v). High correlations (>0.85) was obtained between total carotenoid content and color determination in the UAE oil extracts. UAE vegetable oil extracts enriched with carotenoids from papaya by-products could be useful to formulate new food ingredients based on emulsions with interesting potential health benefits.

Bioaccessibility of Antioxidants in Prickly Pear Fruits Treated with High Hydrostatic Pressure: An Application for Healthier Foods.

High hydrostatic pressure (HHP) is a commercial processing technology which can enhance the health potential of foods by improving the bioaccessibility of their bioactive compounds. Our aim was to study the bioaccessibility and digestive stability of phenolic compounds and betalains in prickly pear fruits (Opuntia ficus-indica L. Mill. var. Pelota and Sanguinos) treated with HHP (100, 350, and 600 MPa; come-up time and 5 min). The effects of HHP on pulps (edible fraction) and peels (sources of potential healthy ingredients) were assessed. In pulps, betanin bioaccessibility increased (+47% to +64%) when treated at 350 MPa/5 min. In HHP-treated pulps, increases in the bioaccessibility of piscidic acid (+67% to +176%) and 4-hydroxybenzoic acid glycoside (+126% to 136%) were also observed. Isorhamnetin glycosides in peels treated at 600 MPa/CUT had higher bioaccessibility (+17% to +126%) than their controls. The effects of HHP on the bioaccessibility of health-promoting compounds are not exclusively governed by extractability increases of antioxidants in the food matrix (direct effects). In this work we found evidence that indirect effects (effects on the food matrix) could also play a role in the increased bioaccessibility of antioxidants in fruits treated with HHP.

In Vitro Antioxidant Capacity of Opuntia spp. Fruits Measured by the LOX-FL Method and its High Sensitivity Towards Betalains.

Current in vitro methodologies neglect or subestimate the contribution of betalains to antioxidant capacity in foods because they do not reflect their in vivo biological mechanisms. In this study, we assessed the sensibility of the lipoxygenase-fluorescein (LOX-FL) method towards betalains, phenolic compounds and ascorbic acid from Opuntia spp. fruits; and (ii) the antioxidant capacity of peel and pulp extracts from Opuntia ficus-indica L. Mill (var. Fresa, Colorada and Blanco) and Opuntia stricta var. Dillenii; by comparing the LOX-FL method to traditional antioxidant methods (ORAC and TEAC). The spectrophotometric monitoring of the LOX-FL reaction avoided interference caused by betalain pigments. Indicaxanthin and betanin showed high antiperoxidative and radical scavenging mechanisms in the LOX-FL assay. O. stricta var. Dillenii tissues the highest antioxidant capacity which correlated with betanin content. ORAC and TEAC antioxidant methods were less sensible towards betalain antioxidant activity. To our knowledge, this is the first time the LOX-FL antioxidant method has been used on betalains and betalain-rich foods.

In Vitro Fecal Fermentation of High Pressure-Treated Fruit Peels Used as Dietary Fiber Sources.

Fruit by-products are being investigated as non-conventional alternative sources of dietary fiber (DF). High hydrostatic pressure (HHP) treatments have been used to modify DF content as well as its technological and physiological functionality. Orange, mango and prickly pear peels untreated (OU, MU and PPU) and HHP-treated at 600 MPa (OP/55 °C and 20 min, MP/22 °C and 10 min, PPP/55 °C and 10 min) were evaluated. Untreated and treated fruit peels were subjected to fecal in vitro fermentations. The neutral sugar composition and linkage glycosidic positions were related to the production of short chain fatty acids (SCFA) resulting from the fermentation of the materials. After HHP-treatments, changes from multibranched sugars to linear sugars were observed. After 24 h of fermentation, OP yielded the highest amount of SCFA followed by PPU and MP (389.4, 282.0 and 204.6 µmol/10 mg DF, respectively). HHP treatment increased the SCFA concentration of orange and mango peel by 7 and 10.3% respectively, compared with the untreated samples after 24 h of fermentation. The results presented herein suggest that fruit peels could be used as good fermentable fiber sources, because they yielded high amounts of SCFA during in vitro fermentations.

Inhibitory potential of prickly pears and their isolated bioactives against digestive enzymes linked to type 2 diabetes and inflammatory response.

BACKGROUND: Prickly pears are potential candidates for the development of low-cost functional foods because they grow with low water requirements in arid regions of the world. They are sources of betalains and phenolic compounds, which have been reported to contribute to human health. The study of the biological activity of different varieties and of their isolated bioactive constitutes is fundamental in the design of functional foods. In this context, our objective is the assessment of the ability of Spanish and Mexican prickly-pear cultivars to inhibit enzymes related to type 2 diabetes and the inflammatory response, and the contribution of their bioactive compounds to their nutra-pharmaceutical potential. RESULTS: Prickly pear peels presented the highest antioxidant activity due to their high isorhamnetin glycoside content. Isorhamnetin glycosides showed significantly higher antioxidant and anti-inflammatory activity than aglycone, particularly isorhamnetin glucosyl-rhamnosyl-pentoside (IG2), which also reported antihyperglycemic activity. Morada, Vigor, and Sanguinos whole fruits exhibited moderate α-amylase inhibition and higher α-glucosidase inhibition, which is ideal for lowering glucose absorption in hyperglycemia management. Sanguinos peels presented the highest anti-inflammatory activity because of their high indicaxanthin content and isorhamnetin glycoside profile. CONCLUSIONS: In the design of prickly pear functional foods, technological processing should prioritize the retention or concentration of these bioactive compounds to preserve (or increase) their natural antioxidant, antihyperglycemic and anti-inflammatory activity. Peels of red and orange varieties should be further evaluated for antioxidant and anti-inflammatory purposes while whole fruits of red and purple varieties could be considered possible candidates for hyperglycemia management. © 2019 Society of Chemical Industry.

In vitro bioaccessibility of individual carotenoids from persimmon (Diospyros kaki, cv. Rojo Brillante) used as an ingredient in a model dairy food.

BACKGROUND: Addition of persimmon fruit, which is highly rich in carotenoids, to dairy products represents an alternative to obtain functional foods. However, carotenoid bioaccessibility is strongly influenced by fat content and food composition. That is why in vitro bioaccessibility of individual carotenoids was evaluated in persimmon-based dairy products formulated with whole (3.6% fat) or skimmed milk (0.25% fat) and different freeze-dried persimmon tissues. RESULTS: Unambiguous identification of seven xanthophylls (neoxanthin, violaxanthin, antheraxanthin, lutein, zeaxanthin, lutein epoxide and ß-cryptoxanthin) and three hydrocarbon carotenes (α-carotene, ß-carotene and lycopene) was achieved using high-performance liquid chromatography with a reverse-phase C-30 column. Total carotenoid content declined up 71% through the digestion process. In vitro bioaccessibility of carotenoids was significantly higher in dairy products formulated with whole milk than those with skimmed milk, representing a difference of more than 21% (in the formulation using persimmon whole fruit as ingredient). Furthermore, addition of whole milk to any type of persimmon tissue significantly improved the bioaccessibility of total provitamin A carotenoids, reaching the highest values (38%) with whole fruit and whole milk. CONCLUSION: The higher fat content in whole milk exerted a significant influence on carotenoid bioaccessibility, especially when using freeze-dried persimmon whole fruit. © 2017 Society of Chemical Industry.

Anti-Obesity Effect of Different Opuntia stricta var. dillenii's Prickly Pear Tissues and Industrial By-Product Extracts in 3T3-L1 Mature Adipocytes.

Opuntia stricta var. dillenii fruit is a source of phytochemicals, such as betalains and phenolic compounds, which may play essential roles in health promotion. The aim of this research was to study the triglyceride-lowering effect of green extracts, obtained from Opuntia stricta var. dillenii fruit (whole fruit, pulp, peel, and industrial by-products (bagasse)) in 3T3-L1 mature adipocytes. The cells were treated on day 12, for 24 h, after the induction of differentiation with the extracts, at doses of 10, 25, 50, or 100 µg/mL. The expression of genes (PCR-RT) and proteins (Western blot) involved in fatty acid synthesis, fatty acid uptake, triglyceride assembly, and triglyceride mobilisation was determined. The fruit pulp extraction yielded the highest levels of betalains, whereas the peel displayed the greatest concentration of phenolic compounds. The extracts from whole fruit, peel and pulp were effective in reducing triglyceride accumulation at doses of 50 µg/mL or higher. Bagasse did not show this effect. The main mechanisms of action underpinning this outcome encompass a reduction in fatty acids synthesis (de novo lipogenesis), thus limiting their availability for triglyceride formation, alongside an increase in triglyceride mobilisation. However, their reliance is contingent upon the specific Opuntia extract.

Encapsulation of Indicaxanthin-Rich Opuntia Green Extracts by Double Emulsions for Improved Stability and Bioaccessibility.

Opuntia ficus-indica var. Colorada fruit is an important source of indicaxanthin, a betalain with antioxidant, anti-inflammatory, and neuromodulatory potential, proven in both in vitro and in vivo models. Other betalains and phenolic compounds with bioactive activities have also been identified in Colorada fruit extracts. These compounds may degrade by their exposure to different environmental factors, so in the present research, two double emulsion systems (W1/O/W2) were elaborated using Tween 20 (TW) and sodium caseinate (SC) as surfactants to encapsulate Colorada fruit pulp extracts, with the aim of enhancing their stability during storage. Encapsulation efficiencies of up to 97.3 ± 2.7%, particle sizes between 236 ± 4 and 3373 ± 64 nm, and zeta potential values of up to ∣46.2∣ ± 0.3 mV were obtained. In addition, the evaluation of the in vitro gastro-intestinal stability and bioaccessibility of the main individual bioactives was carried out by standardized INFOGEST© protocol, obtaining the highest values for the encapsulated extract bioactives in comparison with the non-encapsulated extract (control). Especially, TW double emulsion showed bioaccessibility values of up to 82.8 ± 1.5% for the main bioactives (indicaxanthin, piscidic acid, and isorhamnetin glucoxyl-rhamnosyl-pentoside 2 (IG2)), indicating a promising potential for its use as a functional natural colorant ingredient.

A Healthy Brazil Nut Beverage with Opuntia stricta var. dillenii Green Extract: Beverage Stability and Changes in Bioactives and Antioxidant Activity during Cold Storage.

Plant-based beverages are one of the foods that currently arouse a lot of interest in the population due to their composition with compounds beneficial to health in addition to their being used as milk substitutes for people who suffer from food disorders. Also, their fortification with different nutrients or healthy ingredients with the aim of improving plant-based health potential is actually gaining importance in the food industry. For this reason, the aim of the present investigation was the preparation of a healthy Brazil nut beverage enriched with Opuntia stricta var. dillenii pulp green extracts (ODPs), in order to produce a healthy plant-based beverage with improved nutritional characteristics. The microstructural characterization of the Brazil nut beverage, its stability during cold storage for up to 24 days at 5 °C, the composition of bioactive compounds provided via ODP extract (betalains and phenolic compounds), and their antioxidant activity were evaluated in this study. Green ODP extracts (0.5 and 1 g/100 g beverage) were added to a standardized Brazil nut beverage (reduced fat beverage). The characterization of the bioactive composition (betalains and phenolic compounds) of the elaborated beverage was achieved via HPLC (UV-vis and MS-QT of detection), and the antioxidant activity measurements via ORAC were also carried out. Optical microscopy, particle size, and Z potential analysis was conducted to characterize the structure of the Brazil nut beverages as food emulsions in which ODP extract was added. Most of the bioactive compounds from the green ODP extract added to the beverages showed good retention and remained stable throughout the 24 days of storage at 7 °C, with encapsulation efficiencies ranging from 98.34% to 92.35% for betalains and from 93.67% and 81.20% for phenolic compounds. According to the results of this study, Brazil nut beverage seems to be a healthy and efficient food emulsion system to encapsulate ODP extract rich in betalains and phenolic compounds, with high antioxidant activity, making possible the development of a Brazil nut beverage with improved health potential.

Improvement in the Stability and Bioaccessibility of Carotenoid and Carotenoid Esters from a Papaya By-Product Using O/W Emulsions.

The aim of the present work was to improve the stability and bioaccessibility of carotenoids from green oil extracts obtained from papaya by-products using oil-in-water (O/W) emulsions. The effects of different concentrations of pectin (1%, 2%, and 3%), a high-molecular-size emulsifier, together with Tween 20, a low-molecular-size emulsifier, high-speed homogenization conditions (time: 2, 3, 4, and 5 min; rpm: 9500, 12,000, 14,000, and 16,000 rpm), and high-pressure homogenization (HPH) (100 MPa for five cycles) were evaluated to determine the optimal conditions for obtaining O/W stable emulsions with encapsulated carotenoids. Soybean, sunflower, and coconut oils were used to formulate these O/W emulsions. The bioaccessibility of the main individual encapsulated papaya carotenoids was evaluated using the INFOGEST digestion methodology. In addition, the microstructures (confocal and optical microscopy) of the O/W carotenoid emulsions and their behavior during in vitro digestion phases were studied. Sunflower O/W carotenoid emulsions showed smaller mean particle size, higher negative ζ-potential, and higher viscosity than soybean O/W emulsions. Particle size reduction in the O/W emulsions using the HPH process improved the bioaccessibility of papaya encapsulated carotenoids. In these O/W emulsions, depending on the vegetable oil, lycopene was the carotenoid with the highest bioaccessibility (71-64%), followed by (all-E)-ß-carotene (18%), (all-E)-ß-cryptoxanthin (15%), and (all-E)-ß-cryptoxanthin laurate (7-4%). These results highlight the potential of using green carotenoid papaya extracts to formulate O/W emulsions to enhance carotenoid bioactivity by efficiently preventing degradation and increasing in vitro bioaccessibility.

Oil-Based Double Emulsion Microcarriers for Enhanced Stability and Bioaccessibility of Betalains and Phenolic Compounds from Opuntia stricta var. dillenii Green Extracts.

Opuntia cactus fruit (prickly pear flesh and agricultural residues such as peels and stalks) is an important source of bioactive compounds, including betalains and phenolic compounds. In this work, two double emulsion W1/O/W2 formulations (A and B) were designed to encapsulate green extracts rich in betalains and phenolic compounds obtained from Opuntia stricta var. dillenii (OPD) fruits with the aim of improving their stability and protecting them during the in vitro gastrointestinal digestion process. In addition, the characterization of the double emulsions was studied by microscopy and the evaluation of their physical and physico-chemical parameters. Formulation A, based on Tween 20, showed smaller droplets (1.75 µm) and a higher physical stability than Formulation B, which was achieved with sodium caseinate (29.03 µm). Regarding the encapsulation efficiency of the individual bioactives, betalains showed the highest values (73.7 ± 6.7 to 96.9 ± 3.3%), followed by flavonoids (68.2 ± 5.9 to 95.9 ± 7.7%) and piscidic acid (71 ± 1.3 to 70.2 ± 5.7%) depending on the formulation and the bioactive compound. In vitro digestive stability and bioaccessibility of the individual bioactives increased when extracts were encapsulated for both formulations (67.1 to 253.1%) in comparison with the non-encapsulated ones (30.1 to 64.3%), except for neobetanin. Both formulations could be considered as appropriate microcarrier systems for green OPD extracts, especially formulation A. Further studies need to be conducted about the incorporation of these formulations to develop healthier foods.

Beneficial Effects of Opuntia spp. on Liver Health.

The genus Opuntia spp. includes plants capable of growing in arid, temperate and tropical climates. The vast majority of wild species grow in Mexico, but O. ficus-indica (prickly pear or nopal) is cultivated around the world and it is one of the most studied. This review shows the currently available knowledge concerning the action of O. ficus-indica and other Opuntia species (Opuntia vulgaris, Opuntia robusta, Opuntia streptacantha, Opuntia microdasys, Opuntia dillenii and Opuntia dejecta) on liver health. The available data demonstrate the positive effects of extracts, vinegar, juices or seed oil of the Opuntia genus on the alterations induced in the liver by inadequate feeding patterns or the administration of chemicals. In this regard, the potential beneficial effects of nopal are related to the attenuation of triglyceride accumulation, oxidative stress and/or inflammation. Nevertheless, there is no information concerning the bioactive compound's characterisation in most of these studies; consequently, it is not possible to link the therapeutic effects of these plants to the presence of specific compounds in the nopal extracts. Therefore, further research is needed to confirm if the positive effects observed in animal models are also found in humans, in order to determine whether Opuntia can represent an effective tool to prevent and/or manage hepatic alterations.

Greater bioavailability of xanthophylls compared to carotenes from orange juice (high-pressure processed, pulsed electric field treated, low-temperature pasteurised, and freshly squeezed) in a crossover study in healthy individuals.

This study examined the effect of the intake of orange juice provided freshly squeezed (FS) or processed using low-temperature pasteurisation (LP), high-pressure processing (HPP), or pulsed electric field (PEF) treatment on the serum carotenoid concentrations of 12 healthy individuals, aged 20-32 years, enrolled in a crossover study. Participants were instructed to consume 500 ml of orange juice/day for 14 days. Carotenoid concentrations in the orange juice as well as serum samples retrieved on days 7 and 14 were analysed via HPLC. A significant increase in serum xanthophyll concentrations, but not serum carotenes, was observed, with the highest increase in α- and ß-cryptoxanthin. The processing technologies applied appeared to affect serum carotenoid concentrations, with concentrations being similar in the HPP and FS orange juice types. As high variability in serum carotenoid concentrations was observed, the effect of different technologies on serum carotenoid concentration warrants further studies with larger sample sizes.

Effect of High Hydrostatic Pressure on the Extractability and Bioaccessibility of Carotenoids and Their Esters from Papaya (Carica papaya L.) and Its Impact on Tissue Microstructure.

High hydrostatic pressure (HHP) is a non-thermal technology widely used in the industry to extend food shelf-life and it has been proven to enhance the extractability of secondary metabolites, such as carotenoids, in plant foods. In this study, fresh-cut papaya pulp of varieties (Sweet Mary, Alicia and Eksotika) from the Canary Islands (Spain) were submitted to the HHP process (pressure: 100, 350 and 600 MPa; time: come-up time (CUT) and 5 min) to evaluate, for the first time, individual carotenoid and carotenoid ester extractability and to assess their bioaccessibility using an in vitro simulated gastrointestinal digestion assay, following the standardized INFOGEST® methodology. In addition, changes in papaya pulp microstructure after HHP treatments and during the different phases of the in vitro digestion were evaluated with optical light microscopy. HPLC-DAD (LC-MS/MS (APCI+)) analyses revealed that HHP treatments increased the carotenoid content, obtaining the highest extractability in pulp of the Sweet Mary papaya variety treated at 350 MPa during 5 min (4469 ± 124 µg/100 g fresh weight) which was an increase of 269% in respect to the HHP-untreated control sample. The highest carotenoid extraction value within each papaya variety among all HHP treatments was observed for (all-E)-lycopene, in a range of 98-1302 µg/100 g fresh weight (23-344%). Light micrographs of HHP-treated pulps showed many microstructural changes associated to carotenoid release related to the observed increase in their content. Carotenoids and carotenoid esters of papaya pulp submitted to in vitro digestion showed great stability; however, their bioaccessibility was very low due to the low content of fatty acids in papaya pulp necessary for the micellarization process. Further studies will be required to improve papaya carotenoid and carotenoid ester bioaccessibility.

Carotenoid and Carotenoid Ester Profile and Their Deposition in Plastids in Fruits of New Papaya (Carica papaya L.) Varieties from the Canary Islands.

The carotenoid profile of non-saponified and saponified extracts of different tissues (pulp and peel) of fruits of three new papaya varieties, Sweet Mary, Alicia, and Eksotika, was characterized for the first time, and almost all carotenoid compounds were quantified. Carotenoids and carotenoid esters were analyzed and characterized using HPLC-photo diode array (PDA-MS with atmospheric pressure chemical ionization with positive ion mode (APCI+) with a C30 reversed-phase column. The carotenoid deposition in collenchyma and chlorenchyma cells of papaya pulp and peel tissues was assessed by optical microscopy, confocal laser scanning microscopy, and transmission electron microscopy. The most abundant carotenoids in the fruit of the three papaya varieties (pulp and peel) were (all-E)-lycopene (230.0-421.2 µg/100 g fresh weight), (all-E)-ß-carotene (120.3-233.2 µg/100 g fresh weight), and (all-E)-ß-cryptoxanthin laurate (74.4-223.2 µg/100 g fresh weight. Moreover, high concentrations of (all-E)-lutein (922.5-1381.1 µg/100 g fresh weight) and its esters, such as (all-E)-lutein-3-O-myristate and (all-E)-lutein dimyristate, were found in peel extracts. The optical microscopy study of papaya pulps showed that carotenoid deposition in all papaya varieties, including Maradol, was mainly localized close to the cell walls, showing the presence of some crystalloids and round-shaped structures, with different sizes and distribution due to the different carotenoid content among varieties. No crystalloids or globular depositions were found in any of the peel sections, and no remarkable differences were found in the papaya peel microstructure of the different papaya varieties.

In vitro gastrointestinal stability, bioaccessibility and potential biological activities of betalains and phenolic compounds in cactus berry fruits (Myrtillocactus geometrizans).

Cactus berry (Myrtillocactus geometrizans) is a scarcely studied Mexican wild fruit. These fruits could contribute to reduce the risk of degenerative chronic diseases due to their bioactive profile. The aim of this work was to study the betalains and phenolic profile in cactus berry, their in vitro biological activities and gastrointestinal digestive stability and bioaccessibility. 43 metabolites were identified by HPLC-DAD-ESI-QTOF (8 betaxanthins, 8 betacyanins, 13 flavonoids, 6 phenolic acids). Phyllocactin and Isorhamnetin rhamnosyl-rutinoside (IG2) were the most abundant metabolites (5876 and 396 µg/g dw) which were also bioaccessible (16 and 21%, respectively). Pulps showed higher (p ≤ 0.05) antioxidant activity by the Oxygen Radical Absorbance Capacity (27 mM Trolox equivalents). The anti-hyperglycemic activity was highest (p ≤ 0.05) in peel and pulp tissues (85% α-glucosidase and 8% α-amylase inhibition). An 83% inhibition of hyaluronidase showed high anti-inflammatory activity. Cactus berry fruit should be considered a promising fruit candidate for a sustainable healthy diet.