Lutein bioaccessibility in casein-stabilized emulsions is influenced by the free to acylated carotenoid ratio, but not by the casein aggregation state.

Considering that carotenoids are found acylated to fatty acids in most edible fruits, the influence of the ratio of free to acylated lutein on the hydrolysis extent and bioaccessibility was evaluated by in vitro digestion. For this purpose, for the first time, esterified, free, or a mixture of both carotenoid forms was used in the lipid phase of emulsions stabilized by sodium caseinate (NaCas) and native phosphocaseinate (PPCN). Marigold petals was used as a source of lutein-rich extracts. The emulsions were characterized and the extent of ester hydrolysis, carotenoid recovery, and bioaccessibility were evaluated by LC-DAD-MS/MS. Besides low polydispersity, NaCas and PPCN stabilized emulsions exhibited a constant mean droplet diameter of about 260 and 330 nm, respectively, after 7 days. Caseins were completely digested after the gastric digestion step. Moreover, casein supramolecular structure did not significantly affect carotenoid bioaccessibility. Lutein was majorly found in its free form in all bioaccessible fractions. The carotenoid bioaccessibility increased from 3% to 40% by increasing the percentage of free carotenoids from 0.5 to 100% in the emulsions; but the carotenoid recovery and hydrolysis extent of lutein esters were not affected. In conclusion, emulsion-based systems for carotenoid delivery stabilized either by NaCas or PPCN provided similar carotenoid bioaccessibility. Furthermore, bioaccessibility was inversely dependent on the overall hydrophobicity of the carotenoid extract. Our results suggest that the low bioaccessibility of esterified carotenoids was a consequence of their limited hydrolysis extent. This study provides information that may help design emulsion-based systems stabilized by food protein as a vehicle for carotenoids.

A guide for the evaluation of in vitro bioaccessibility of carotenoids.

Ongoing efforts to improve the nutritional content of carotenoids in food sources to solve global health problems must also consider bioaccessibility and bioavailability of carotenoids. Carotenoid bioaccessibility encompasses carotenoid release from the food matrix, solubilization in lipid emulsion droplets, and transfer to mixed micelles for further absorption into the intestine. Bioavailability refers to the fraction of carotenoids which enters into circulation. To understand the mechanisms controlling bioaccessibility, in vitro digestion methods have been developed and are a useful approach to overcome some of the disadvantages associated with in vivo protocols. The INFOGEST protocol comprises three phases (oral, gastric and intestinal) to simulate adult human digestion of foods, as an international consensus for the conditions of static in vitro digestion. The last steps in any digestion protocol involve isolation of the target food compounds and subsequent analysis, steps not included in the INFOGEST protocol because they may differ depending on specific physical-chemical characteristics and quantification analysis of the target analyte. In this chapter, we describe the INFOGEST protocol which has been adapted for the analysis of carotenoid bioaccessibility, including determination and calculation of enzyme activities according to INFOGEST guidelines, removal of undigested material, collection of mixed micelles containing carotenoids, extraction, storage and subsequent identification and quantification by liquid chromatography analysis of both free and acylated carotenoids.

New cyclopeptide alkaloid of Condalia buxifolia and the absolute stereochemistry of Condaline A.

During the course of a study of Condalia buxifolia (Rhamnaceae), one new cyclopeptide alkaloid condaline B (1), together with six known cyclopeptide alkaloids, condaline A (2), and the scutianines B (3), - D (4) and -E (5), frangulanine (6), and 3,4,28-tris-epi-scutianene N (7), were isolated from the rind bark of Condalia buxifolia. Their structures have been confirmed through spectroscopic analyses such as 1D and 2D NMR experiments. The absolute stereochemistry of condaline A (2), was elucidated by X-ray crystal structure determination of its HI salt. In addition, condaline B (1) was obtained synthetically through a structural transformation of condaline A. Meanwhile, the crude methanol extract, the basic ether fraction, and alkaloids 1-7 were tested against various strains of Gram-positive and Gram-negative bacteria and fungus, showing promising antimicrobial activity.

Is noninvasive neurally adjusted ventilatory assistance (NIV-NAVA) an alternative to NCPAP in preventing extubation failure in preterm infants?

BACKGROUND: Prolonged use of mechanical ventilation is associated with some complications as high mortality and high morbidities as bronchopulmonary dysplasia, ventilator-associated pneumonia, and pneumothorax. However, extubation failure in preterm infants is still high (40-60%) in very low birth weight infants (VLBW). Noninvasive neurally adjusted ventilatory assistance (NIV-NAVA) is triggered by the diaphragmatic electrical activity through a nasogastric tube that synchronizes patient/ventilator respiration, cycle by cycle effectively shortening the assisted cycle trigger and the degree of ventilatory assistance, optimizing the effects of intermittent inspiratory pressure on nasal continuous positive airway pressure (NCPAP). This study aims to compare reintubation rates until 72 h after extubation in preterm infants of high risk for reintubation using NIV-NAVA or NCPAP. METHODS: A retrospective study of chart review data collection was performed in a private tertiary hospital. The study was approved by the local institutional Ethics Committee. We included infants considered at high risk of reintubation (BW < 1000 grams; use of invasive mechanical ventilation (IMV) for at least 7 days; or previous extubation failure episode) and compared the two groups according to the type of respiratory support after extubation: (1) NCPAP (n = 32); or (2) NIV-NAVA (n = 17). Demographics data were collected, the primary outcome was reintubation rate until 72 h after extubation. Secondary outcome was time to reintubation, BPD rate, IVH grade ≥ III, pneumothorax and death. RESULTS: There was no difference between both groups in demographic data. The reintubation rate decreased significantly in the NIV-NAVA group compared to NCPAP (50.0-11.7, p < .02) despite the significantly higher length of invasive mechanical ventilation (IMV) before extubation attempt in NIV-NAVA group (12.4 versus 5.5 days, p < .04). There was no difference between both groups in secondary outcomes. CONCLUSIONS: In this small retrospective cohort study, the use of NIV-NAVA as postextubation strategy was effective in reducing extubation failure within 72 h in preterm infants when compared to traditional NCPAP.

HPLC-PDA-MS/MS as a strategy to characterize and quantify natural pigments from microalgae.

Interest in pigment composition of microalgae species is growing as new natural pigments sources are being sought. However, we still have a limited number of species of microalgae exploited to obtain these compounds. Considering these facts, the detailed composition of carotenoids and chlorophylls of two species of green microalgae (Chlorella sorokiniana and Scenedesmus bijuga) were determined for the first time by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-PDA-MS/MS). A total of 17 different carotenoids were separated in all the extracts. Most of the carotenoids present in the two microalgae species are xanthophylls. C. sorokiniana presented 11 carotenoids (1408.46 µg g-1), and S. bijuga showed 16 carotenoids (1195.75 µg g-1). The main carotenoids detected in the two microalgae were all-trans-lutein and all-trans-ß-carotene. All-trans-lutein was substantially higher in C. sorokiniana (59.01%), whereas all-trans-ß-carotene was detected in higher quantitative values in S. bijuga (13.88%). Seven chlorophyll compounds were identified in both strains with different proportions in each species. Concentrations of chlorophyll representing 7.6% and 10.2% of the composition of the compounds present in the biomass of C. sorokiniana and S. bijuga, respectively. Relevant chlorophyll compounds are reported for the first time in these strains. The data obtained provide significant insights for microalgae pigment composition databases.

Ionic liquid associated with ultrasonic-assisted extraction: A new approach to obtain carotenoids from orange peel.

The aim of this study was to develop a new method for carotenoid extraction from orange peel, using ionic liquid (IL) to replace conventional organic solvents, assisted by ultrasound. Four different IL were tested: 1-butyl-3-methylimidazolium chloride ([BMIM][Cl]), 1-n-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]), 1-n-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]), and 1-hexyl-3-methylimidazolium chloride ([HMIM][Cl]). Response surface methodology was applied in order to optimize the carotenoid extraction conditions, and Amberlite XAD-7HP resin was used to separate the carotenoids from the IL, allowing their recovery. Determination of carotenoids was carried out by high-performance liquid chromatography coupled to photodiode array and mass spectrometry detectors (HPLC-DAD-MSn). Thermal stability at different temperatures (60 °C and 90 °C) and peroxyl radical scavenging activity of the carotenoid extracts obtained with acetone and IL were evaluated. [BMIM][Cl] was the most effective IL, leading to a total carotenoid content of 32.08 ±â€¯2.05 µg/g, while 7.88 ±â€¯0.59 µg/g of dry matter was obtained by acetone extraction. IL and carotenoid recoveries using XAD-7HP resin were in the range of 59.5-63.8% and 52.2-58.7%, respectively. A carotenoid extract was successfully obtained with IL, finally isolated just by using ethanol, besides being more stable and presenting higher antioxidant activity than that obtained with acetone.

Chemical constituents from Valeriana polystachya Smith and evaluation of their effects on the acetylcholinesterase and prolyl oligopeptidase activities.

Two new iridoids (1-2) and a new decomposition product of valepotriates (3), together with fifteen known compounds (4-18) were isolated from the roots and rhizomes of Valeriana polystachya Smith, a native species from the Pampa Biome. Their structures were elucidated by means of NMR spectroscopy, mass spectrometry and optical rotation. The structures of 3 and 18 were further confirmed by single crystal X-ray diffraction analysis. In the group of the isolated compounds, 6ß-hydroxysitostenone, hydroxymaltol and isovillosol were isolated from the Valeriana genus for the first time. The extracts and isolated compounds were evaluated for their in vitro activities against acetylcholinesterase (AChE) and prolyloligopeptidase (POP). Compounds 7, 9 and 11 showed weak inhibitory activity against AChE, while 3 and 5 displayed exceptional POP inhibitory activity, with IC50 values of 5.3 ±â€¯0.07 and 7.9 ±â€¯0.4 µM, respectively.

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.

Carotenoid profile of three microalgae/cyanobacteria species with peroxyl radical scavenger capacity.

Carotenoids from cyanobacteria Aphanothece microscopica Nageli and green microalgae Chlorella vulgaris and Scenedesmus obliquus were identified. The total carotenoid content, based on dry weight of biomass, of A. microscopica Nägeli, C. vulgaris and S. obliquus were 1398.88µg/g, 1977.02µg/g and 2650.70µg/g, respectively. A total of 23 different carotenoids were separated in all the extracts, the major ones being all-trans-ß-carotene (29.3%) and all-trans-lutein (28.1%) in Scenedesmus; all-trans-echinenone (22.8%) and all-trans-ß-carotene (17.7%) in Chlorella; all-trans-echinenone (28.3%) and all-trans-ß-carotene (26.2%) in Aphanothece. The carotenoid extracts were shown to be a potent scavenger of peroxyl radical, with values of 31.1 (Chlorella), 14.0 (Scenedesmus) and 7.3 (Aphanothece) times more potent than α-tocopherol.

Identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale using ultrasound-assisted extraction.

The identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale was determined. The ultrasound-assisted extraction was utilized for obtaining the chlorophyll compounds from biomass. A total of eleven molecules were separated in microalgae chlorophyll extract, with pheophytin a' (371µg·g-1) and chlorophyll a (159.3µg·g-1) as the major ones. The chlorophyll extract was shown to be a potent scavenger of peroxyl radical, being almost 200 times more potent than α-tocopherol. These facts suggest the microalgae Phormidium autumnale as potential source of bioactive tetrapyrrole compounds.

Effect of Pasteurization on Flavonoids and Carotenoids in Citrus sinensis (L.) Osbeck cv. 'Cara Cara' and 'Bahia' Juices.

Orange juice is considered an excellent dietary source of several bioactive compounds with beneficial properties for human health. Citrus sinensis Osbeck cv. 'Cara Cara' is a bud mutation originated from 'Washington' navel orange, also known as 'Bahia' navel orange. The ascorbic acid, flavonoid, and carotenoid contents in pasteurized and nonpasteurized Bahia and Cara Cara juices using two LC-MS/MS platforms were investigated. Higher ascorbic acid content was observed in Bahia compared to Cara Cara in both pasteurized and nonpasteurized juices. Total flavanones content as well as hesperidin levels were higher in Cara Cara with respect to Bahia pasteurized juice. Cara Cara was also characterized by a significantly higher and diversified carotenoid content compared to Bahia juice with a mixture of (Z)-isomers of lycopene, all-E-ß-carotene, phytoene, and phytofluene isomers accounting for the highest carotenoid proportion. The exceptionally high carotenoid content of Cara Cara may be particularly interesting for nutritional or functional studies of uncommon carotenes in a citrus food matrix.

Composition by LC-MS/MS of New Carotenoid Esters in Mango and Citrus.

Interest in the composition of carotenoid esters of fruits is growing because esterification may affect their bioavailability. Thus, the aim was to provide a detailed identification of carotenoid esters in citrus and mango. Orange cv. 'Valencia' and cv. 'Pera' presented 9 free carotenoids, 38 monoesters, and 60 diesters. Violaxanthin and luteoxanthin derivatives were the major ones, followed by antheraxanthin, lutein, zeaxanthin, ß-cryptoxanthin, and zeinoxanthin esters, many of them reported for the first time in orange pulp. The carotenoid ester composition of tangor cv. 'Murcott', reported for the first time, showed 8 free carotenoids, 34 monoesters, and 33 diesters, with ß-cryptoxanthin esters as major compounds, followed by violaxanthin and zeaxanthin esters. In citrus, carotenoids were acylated mainly with capric, lauric, myristic, myristoleic, palmitic, palmitoleic, and oleic acids. In mango, 5 free carotenoids, 2 monoesters, and 19 diesters were identified, from which many violaxanthin and neoxanthin esters were reported for the first time.

Changes in antioxidant capacity and colour associated with the formation of ß-carotene epoxides and oxidative cleavage derivatives.

In this study HPLC-DAD-MS/MS was applied for the identification of compounds derived from (all-E)-ß-carotene following epoxidation and oxidative cleavage. The consequences on the CIELAB colour parameters and antioxidant capacity (AC) were also evaluated. Five apocarotenoids, three secocarotenoids, seven Z isomers and two epoxides were detected as a result of the oxidative cleavage. Four epoxides and three Z isomers were detected as a consequence of the epoxidation reaction. Some compounds were detected for the first time as a result of oxidation reactions. Both treatments led to a marked decrease in b(∗) and Cab(∗) values, indicating that these colour parameters can be used for the rapid assessment of ß-carotene oxidation. The oxidative cleavage of ß-carotene resulted in increased capacity to both scavenge ABTS(+) and quench singlet oxygen. These results suggest that the study of the AC of these oxidative derivatives and their possible usefulness as food ingredients deserves further attention.

Metabolomic investigation of human diseases biomarkers by CE and LC coupled to MS.

Metabolomics is one of the most recent trends in the "omics" era that investigates the end products of an organism activity, that is, all metabolites in a biological system, which are small molecules (less than 1000 Da) from different chemical classes. Metabolomics represents a tool to assess the biochemical activity of a living system through the analysis of substrates and products processed during the metabolism. The analysis of the metabolic profile (nontargeted analysis, i.e. a comparison between samples profiles of individuals) and of specific metabolites (targeted analysis, which quantifies a selected group of metabolites) in biological samples provides an insight into the metabolic state and the biochemical processes of the organism and, therefore, may indicate the onset and the stage of different diseases. An early and accurate diagnosis is essential for successful treatment and probable cure of most illnesses; hence, the investigation of metabolites as disease biomarkers has increased considerably in recent years. This review aims to present the most relevant works that address the nontargeted and targeted analysis of metabolites in different diseases for the past 10 years, including kidney and neurological disorders, cardiovascular diseases, diabetes, and cancer, using CE and LC coupled with the accurate detection of mass spectroscopy.

Carotenoids and phenolic compounds from Solanum sessiliflorum, an unexploited Amazonian fruit, and their scavenging capacities against reactive oxygen and nitrogen species.

The composition of carotenoids and phenolic compounds from mana-cubiu (Solanum sessiliflorum), a fruit native to Amazonia, was determined by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-DAD-MS(n)). The antioxidant capacities of the hydrophilic and carotenoid extracts against some reactive oxygen (ROO(•), H(2)O(2), HOCl, and HO(•)) and nitrogen (ONOO(-)) species were also determined. Seventeen carotenoids and three phenolic compounds were found in mana-cubiu. The major carotenoids were (all-E)-ß-carotene (7.15 µg/g of dry weight) and (all-E)-lutein (2.41 µg/g of dry weight). The 5-caffeoylquinic acid (1351 µg/g of dry weight) was the major phenolic compound, representing more than 78% (w/w) of the total phenolic compounds. Moreover, two dihydrocaffeoyl spermidines were found in the hydrophilic extract. Both mana-cubiu extracts were able to scavenge all the tested reactive species. The carotenoid extract was shown to be a potent scavenger of peroxyl radical, while the hydrophilic extract was a potent hydrogen peroxide and hypochlorous acid scavenger.

Soybean extracts enriched with free isoflavones promote nitric oxide synthesis and affect the proliferation of breast adenocarcinoma cells

Although soybean isoflavones naturally accumulate in their conjugated forms, the beneficial effects on human health of soybean-containing foods have been credited to their aglycone forms. In the present study we analyzed the effects of a chemical agent, sodium nitroprusside (SNP), in eliciting the exudation of non-conjugated isoflavones from intact soybean seeds, embrionary axes and cotyledons. The isoflavones in the exudates were determined by high performance liquid chromatography and mass spectrometry. The effect of the exudates on the emission of nitric oxide (NO) and on the proliferation of breast carcinoma cells (MCF-7) was also evaluated. SNP elicitation increased the production of the aglycone forms dose- and time-dependently. Exudates of embrionary axes and cotyledons stimulated NO emission and showed biphasic effects on viability of MCF-7 cells. At lower concentrations both extracts presented proliferative effects (10-25%), and at higher concentrations inhibited (15%) cell proliferation. The biphasic effect might be due to the action of isoflavone aglycones in activating estrogen receptors which in turn stimulate the production of NO. Overall, the results suggest that soybean extracts enriched in isoflavone aglycones by elicitation with SNP could be exploited as a functional ingredient in the food industry.

Microcapsules containing antioxidant molecules as scavengers of reactive oxygen and nitrogen species.

The antioxidant capacities of gum arabic and maltodextrin microcapsules containing antioxidant molecules (trolox, α-tocopherol, ß-carotene, apo-8'-carotenal and apo-12'-carotenal) against reactive oxygen and nitrogen species were evaluated. The scavenging capacities were influenced by the wall material, the reactive species, namely ROO(), H(2)O(2), HO(), HOCl and ONOO(-), and the antioxidant molecule. In general, a more pronounced enhancement of the antioxidant capacity due to incorporation of antioxidant molecules was observed in gum arabic microcapsules. The empty microcapsules showed capacity to scavenge all the studied ROS and RNS, being gum arabic a more potent antioxidant than maltodextrin. Apo-8'-carotenal incorporation promoted the highest increase in the scavenging capacities among the evaluated antioxidants, varying from 50% to 132% and from 39% to 85% for gum arabic and maltodextrin microcapsules, respectively, suggesting that this carotenoid presented the best balance between the molecule localization inside the microcapsules and the reactivity against the specific reactive species.

Scavenging capacity of marine carotenoids against reactive oxygen and nitrogen species in a membrane-mimicking system.

Carotenoid intake has been associated with the decrease of the incidence of some chronic diseases by minimizing the in vivo oxidative damages induced by reactive oxygen (ROS) and nitrogen species (RNS). The carotenoids are well-known singlet oxygen quenchers; however, their capacity to scavenge other reactive species, such as peroxyl radical (ROO•, hydroxyl radical (HO•), hypochlorous acid (HOCl) and anion peroxynitrite (ONOO⁻), still needs to be more extensively studied, especially using membrane-mimicking systems, such as liposomes. Moreover, the identification of carotenoids possessing high antioxidant capacity can lead to new alternatives of drugs or nutritional supplements for prophylaxis or therapy of pathological conditions related to oxidative damages, such as cardiovascular diseases. The capacity to scavenge ROO•, HO•, HOCl and ONOO⁻ of seven carotenoids found in marine organisms was determined in liposomes based on the fluorescence loss of a fluorescent lipid (C11-BODIPY58¹/59¹) due to its oxidation by these reactive species. The carotenoid-bearing hydroxyl groups were generally more potent ROS scavengers than the carotenes, whilst ß-carotene was the most efficient ONOO⁻ scavenger. The role of astaxanthin as an antioxidant should be highlighted, since it was a more potent scavenger of ROO•, HOCl and ONOO⁻ than α-tocopherol.

Development of a novel micro-assay for evaluation of peroxyl radical scavenger capacity: application to carotenoids and structure-activity relationship.

A micro-assay was developed and validated, using a microplate reader in 96-well format, C(11)-BODIPY(581/591) as fluorescent probe and AIBN as ROO() generator. The structure-activity relationship was established for 15 carotenoid standards, indicating that the opening of the ß-ionone ring and the increase of chromophore extension in the carotenoid structure were the major factors leading to the increase of ROO() scavenging capacity. The values for ROO() scavenging capacity were calculated using α-tocopherol as reference compound. Among the studied carotenoids, all-trans-lycopene was the most efficient ROO() scavenger (8.67±0.74) followed by all-trans-astaxanthin (6.50±0.62). All the carotenoids showed to be more effective ROO() scavengers than α-tocopherol and some hydrophilic compounds. Finally, the method was successfully applied to assay the ROO() scavenging capacity of carotenoid extracts from two Amazonian fruits, peach palm (7.83±0.21) and mamey (6.90±0.44).

Identification of carotenoids with high antioxidant capacity produced by extremophile microorganisms.

In this study, the carotenoids produced by the extremophile microorganisms Halococcus morrhuae, Halobacterium salinarium and Thermus filiformis were separated and identified by high-performance liquid chromatography connected to a diode array detector and a tandem mass spectrometer. The in vitro scavenging capacity of the carotenoid extracts against radical and non-radical species was evaluated. In halophilic microorganisms, the following carotenoids were identified: bacterioruberin, bisanhydrobacterioruberin, trisanhydrobacterioruberin and their derivatives. In the thermophilic bacterium, the carotenoids all-trans-zeaxanthin, zeaxanthin monoglucoside, thermozeaxanthins and thermobiszeaxanthins were identified. The antioxidant capacities of the carotenoid extracts of H. morrhuae (trolox equivalent antioxidant capacity = 5.07 and IC(50) = 0.85 µg mL(-1)) and H. salinarium (trolox equivalent antioxidant capacity = 5.28 and IC(50) = 0.84 µg mL(-1)) were similar and higher than those of the bacterium T. filiformis (trolox equivalent antioxidant capacity = 2.87 and IC(50) = 2.41 µg mL(-1)). This difference is related to the presence of acyclic carotenoids with both large numbers of conjugated double bounds and of hydroxyl groups in the major carotenoid of the halophilic microorganisms.