Changes in phytochemical composition, bioactivity and in vitro digestibility of guayusa leaves (Ilex guayusa Loes.) in different ripening stages.

BACKGROUND: Guayusa (Ilex guayusa Loes.) leaves, native of the Ecuadorian Amazon, are popularly used for preparing teas. This study aimed to assess the influence of leaf age on the phenolic compounds and carotenoids and the bioactivity and digestibility (in vitro) of aqueous and hydroalcoholic leaf extracts. RESULTS: In total, 14 phenolic compounds were identified and quantified. Chlorogenic acid and quercetin-3-O-hexose were the main representatives of the hydroxycinnamic acids and flavonols respectively. Seven carotenoids were quantified, lutein being the main compound. Ripening affected phenolic content significantly, but there was no significant difference in carotenoid content. Antioxidant capacity, measured by the DPPH• method, was also significantly affected by leaf age. The measurement of in vitro digestibility showed a decrease in phenolic content (59%) as well as antioxidant capacity, measured by the ABTS•+ method, in comparison with initial conditions of the guayusa infusion. Antibacterial and anti-inflammatory activities were assayed with young leaves owing to their higher phenolic contents. Guayusa did not show any antibacterial activity against Escherichia coli ATCC 25922 or Staphylococcus aureus ATCC 25923. Finally, the hydroalcoholic and aqueous extracts exhibited high in vitro anti-inflammatory activity (>65%). CONCLUSION: Young guayusa leaves have potential applications as a functional ingredient in food and pharmaceutical industries. © 2017 Society of Chemical Industry.

Banana Passion Fruit (Passiflora mollissima (Kunth) L.H. Bailey): Microencapsulation, Phytochemical Composition and Antioxidant Capacity.

Passiflora mollissima (Kunth) L.H. Bailey is an exotic fruit native to South America, known as taxo in Ecuador. This paper characterizes its flavonoid and carotenoid composition and antioxidant capacity and evaluates the effect of the spray-drying process on its phytochemical composition and antioxidant capacity. A total of 18 flavonoid compounds, nine proanthocyanidins and nine flavan-3-ol monomers, were identified and quantified. Glycosides of (epi)-afzelechin stood out as the most abundant flavonoid. Three carotenoids were identified, with ß-carotene having the highest concentration. The DPPH· and ORAC assay methods indicated a high antioxidant capacity. Furthermore, the bioactive content showed a positive and direct correlation with antioxidant capacity. On the other hand, the spray-drying process produced a stable phytochemical composition and antioxidant activity of taxo. These results demonstrate the potential applicability of microencapsulated taxo as a functional ingredient in the food industry.

Guayusa (Ilex guayusa L.) new tea: phenolic and carotenoid composition and antioxidant capacity.

BACKGROUND: Guayusa (Ilex guayusa Loes) is an evergreen tree native of South America that grows particularly in the upper Amazon region of Ecuador. For its health benefits, it has been cultivated and consumed since ancient times by Amazon indigenous tribes. RESULTS: A total of 14 phenolic compounds were identified and quantified. Chlorogenic acid and quercetin-3-O-hexose were the main representatives of the hydroxycinnamic acids and flavonols, respectively. Five carotenoids were identified, showing lutein the highest concentration. Guayusa leaves revealed high antioxidant capacity determined by two analytical methods, DPPH and ORAC. The industrial processing applied to the leaves modified the composition of bioactive compounds and antioxidant capacity of guayusa. In general, blanched guayusa retained the concentration of phenolic compounds and some carotenoids and similar antioxidant capacity as untreated green leaves. In contrast, fermentation reduced the content of bioactive compounds and showed the lowest antioxidant capacity. CONCLUSION: Therefore, blanched guayusa has potential for product development as a functional ingredient in the food industry. © 2017 Society of Chemical Industry.

A comprehensive review on the colorless carotenoids phytoene and phytofluene.

Carotenoids and their derivatives are versatile isoprenoids involved in many varied actions, hence their importance in the agri-food industry, nutrition, health and other fields. All carotenoids are derived from the colorless carotenes phytoene and phytofluene, which are oddities among carotenoids due to their distinct chemical structure. They occur together with lycopene in tomato and other lycopene-containing foods. Furthermore, they are also present in frequently consumed products like oranges and carrots, among others. The intake of phytoene plus phytofluene has been shown to be higher than that of lycopene and other carotenoids in Luxembourg. This is likely to be common in other countries. However, they are not included in food carotenoid databases, hence they have not been linked to health benefits in epidemiological studies. Interestingly, there are evidences in vitro, animal models and humans indicating that they may provide health benefits. In this sense, the study of these colorless carotenes in the context of food science, nutrition and health should be further encouraged. In this work, we review much of the existing knowledge concerning their chemical characteristics, physico-chemical properties, analysis, distribution in foods, bioavailability and likely biological activities.

Towards more sustainable cooking practices to increase the bioaccessibility of colourless and provitamin A carotenoids in cooked carrots.

The effect of different cooking methods (boiling, baking, steaming and microwaving) on the colour and texture of carrots, as well as on the bioaccessibility of carotenoids, was investigated in order to identify the more "sustainable cooking" methods. Cooking resulted in statistically significant increases in total carotenoid bioaccessibility, both with intensity and duration of treatments. In particular, significant increases in carotenoid bioaccessible content (CBC) were observed, ranging from 6.03-fold (microwave) to 8.90-fold (baking) for the most intense cooking conditions tested. Although the relative concentration of the colourless carotenoids (phytoene and phytofluene) in raw carrots is lower than that of provitamins A α- and ß-carotene, the bioaccessible content of the colourless ones is much higher. From an energy consumption standpoint and considering samples with the same tenderness, the highest CBC values per kWh decreased in the order microwaving > baking > water cooking > steaming. Our findings are important to help combat vitamin A deficiency since increases of up to ∼40-fold and ∼70-fold in the CBCs of the vitamin A precursors α- and ß-carotene, respectively, were observed. These results provide a basis for defining "sustainable cooking" as "cooking practices that optimize intensity, duration and other parameters leading to a more efficient use of energy to maximize the bioavailability of nutrients and other beneficial food components (such as bioactives) while ensuring food appeal and safety".

Assessment of in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage.

Mandarine juice is one of the richest sources of ß-cryptoxanthin and flavonoids, which have been positively associated with bone mineral density. Carotenoids are lipophilic isoprenoid compounds with a complex absorption process that can be affected by different factors. In this study, we have evaluated the effect of the food matrix on the in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage (MMB). MMBs were formulated with mandarine juice and different dairy products to achieve three fat levels (0.2%, 1.7% and 3.2%) and three calcium levels (120, 310 and 500 mg Ca2+ per 100 ml). The bioaccessibility was evaluated using a harmonised in vitro digestion method. The results showed that the content of milk fat increased the bioaccessibility in vitro of phenolic compounds (p < 0.05), while a moderate fat level (1.7%) resulted in the highest bioaccessibility for bioactive carotenoids. On the other hand, calcium fortification at the highest level (500 mg Ca2+ per 100 mL) decreased the bioaccessibility of bioactive carotenoids from 76% to 43% (66% for the major ß-cryptoxanthin) compared to the lower calcium fortification level (120 mg Ca2+ per 100 mL). The bioaccessibility of hesperidin, the main flavanone in mandarine juice, was significantly (p < 0.05) reduced in the MMB with the highest calcium level. The bioaccessibility of carotenoids and phenolic compounds is affected by fat and calcium levels. When formulating functional beverages, the impact of the formulation should be carefully considered to optimize the bioaccessibility of the bioactive compounds.

Assessment of Food Sources and the Intake of the Colourless Carotenoids Phytoene and Phytofluene in Spain.

Phytoene (PT) and phytofluene (PTF), colorless carotenoids, have largely been ignored in food science studies, food technology, and nutrition. However, they are present in commonly consumed foods and may have health-promotion effects and possible uses as cosmetics. The goal of this study is to assess the most important food sources of PT and PTF and their dietary intakes in a representative sample of the adult Spanish population. A total of 62 food samples were analyzed (58 fruit and vegetables; seven items with different varieties/color) and carotenoid data of four foods (three fruits and one processed food) were compiled. PT concentration was higher than that of PTF in all the foods analyzed. The highest PT content was found in carrot, apricot, commercial tomato juice, and orange (7.3, 2.8, 2.0, and 1.1 mg/100 g, respectively). The highest PTF level was detected in carrots, commercial tomato sauce and canned tomato, apricot, and orange juice (1.7, 1.2, 1.0, 0.6, and 0.04 mg/100 g, respectively). The daily intakes of PT and PTF were 1.89 and 0.47 mg/person/day, respectively. The major contributors to the dietary intake of PT (98%) and PTF (73%) were: carrot, tomato, orange/orange juice, apricot, and watermelon. PT and PTF are mainly supplied by vegetables (81% and 69%, respectively). Considering the color of the edible part of the foods analyzed (fruit, vegetables, sauces, and beverages), the major contributor to the daily intake of PT and PTF (about 98%) were of red/orange color.

Distribution of Polyphenolic and Isoprenoid Compounds and Biological Activity Differences between in the Fruit Skin + Pulp, Seeds, and Leaves of New Biotypes of Elaeagnusmultiflora Thunb.

The purpose of this study was to determine the distribution of polyphenolic and isoprenoid compounds and organic acids in the fruit skin + pulp, seeds, and leaves of six new biotypes of Elaeagnus multiflora Thunb., as well as their in vitro biological potency. The polyphenols and isoprenoids were determined with UPLC-PDA-MS/MS (ultra-performance liquid chromatography coupled to photodiode array detection and electrospray ionization tandem mass spectrometry) and RRLC-MS/MS (rapid resolution liquid chromatography/tandem mass spectrometry) methods, the organic acid with HPLC-RID (high-performance liquid chromatography coupled to a Refractive Index Detector), and the antioxidant capacity using ABTS and FRAP assays. Enzymatic activity was established as the ability to inhibit α-amylase, α-glucosidase, and pancreatic lipase. Owing to such an effective technique, 88 compounds were recorded, with 17 polyphenolic compounds and 3 isoprenoids identified for the first time in the seeds and leaves of cherry silverberry. In total, 55 compounds were identified in the leaves, 36 in the seeds, and 31 in the fruit skin + pulp. The predominant polyphenol was polymeric procyanidin (66-95% of total polyphenolics), whereas the predominant isoprenoids were chlorophyll b and (all-E)-lycopene. The results of our work noted that there are significant differences in the profiles of several secondary metabolites between the analyzed parts of the plant, and depending on the need, the compounds can be used to develop different innovative food or cosmetic products.

Applications of Visible Spectroscopy and Color Measurements in the Assessments of Carotenoid Levels in Foods.

The color of the food can be evaluated visually or instrumentally. The instrumental measurement allows to measure the color objectively and can be carried out by different techniques, most of them fast, cheap, and simple. Carotenoids are responsible for the color of many plants and foods. The measurement of color can be an effective tool to determine indirectly the concentration of carotenoids in plants and foods. In this chapter, we describe both traditional and recent methodologies for color measurement.

In Vitro Biological Activities of Fruits and Leaves of Elaeagnus multiflora Thunb. and Their Isoprenoids and Polyphenolics Profile.

The objective of this study was in-depth identification of carotenoids and polyphenolic compounds in leaves and fruits of Elaeagnus multiflora Thunb. An additional aim was to assay their antioxidant and in vitro biological activities (the ability to inhibit pancreatic lipase, α-amylase, and α-glucosidase activity) of two cultivars: 'Sweet Scarlet' and 'Jahidka'. Study results showed the presence of 70 bioactive compounds, including 20 isoprenoids and 50 polyphenols. The profile of identified bioactive compounds had not been examined in this respect until now. The total carotenoid, chlorophyll, and polyphenol levels and antioxidant activity of the foliar samples were virtually identical in both cultivars and clearly higher relative to those in the fruits. On the other hand, the ability to inhibit pancreatic lipase, α-amylase, and α-glucosidase activity of the fruits was clearly higher as compared to the leaves. The highest amount of phenolic acids, flavonols, and polymeric procyanidins was in the 'Sweet Scarlet' for fruit and leaves, while the highest amount of chlorophylls and carotenoids was in the 'Jahidka'. The inhibition of α-amylase, α-glucosidase, and pancreatic lipase activities appeared to be better correlated with the carotenoid content, which warrants further studies of the possible anti-diabetic and anti-obesity actions of the major carotenoids found in the fruits (lycopene, phytoene, and lutein). In addition, strong correlation between antioxidant activity and phenols of E. multiflora Thunb. components can be effective in removing reactive oxygen species. The results of our study show that both the fruits and leaves of E. multiflora Thunb. can be important for health promotion through the diet and for innovating in the industry of functional food and (nutri)cosmetics.

Influence of high pressure homogenization and pasteurization on the in vitro bioaccessibility of carotenoids and flavonoids in orange juice.

Production of high-quality healthy foods through sustainable methodologies is an urgent necessity. High pressure homogenization (HPH) is an interesting alternative to obtain premium citrus juices, but its effects on bioactive compounds are unclear. There was studied the influence of HPH (150 MPa) and pasteurization (92 °C for 30 s and 85 °C for 15 s) processing on physicochemical properties and in vitro bioaccessibility of carotenoids and flavonoids in orange juices. Regarding fresh juice, physicochemical properties of samples remained unchanged although cloudiness was improved by homogenization. Pasteurization did not affect total carotenoids content and retinol activity equivalents (RAE) of juices whereas homogenization yielded a significant reduction (1.37 and 1.35-fold, respectively). Interestingly, particle size reduction from homogenization drastically enhanced (about 5-fold) bioaccessibility of carotenoids including hardly bioaccessible epoxycarotenoids, finding unaltered rates in pasteurized samples. Bioaccessibility of flavonoids was constant in all cases. Results can promote HPH as an efficient option to obtain health-enhanced foods.

Characterization of Andean Blueberry in Bioactive Compounds, Evaluation of Biological Properties, and In Vitro Bioaccessibility.

The aim of this study was to evaluate Andean blueberries (Vaccinium floribundum Kunth) from Ecuador as a potential functional ingredient for the food and pharmaceutical industries. The analysis of bioactive compounds by HPLC-DAD-MSn determined a high content of (poly)phenols, mainly anthocyanins, and the presence of the carotenoid lutein. Regarding its biological properties, Andean blueberry did not show toxicity by the zebrafish embryogenesis test, showing also a lack of the antinutrients lectins. Moreover, the results of in vitro and in vivo antioxidant capacity evaluation suggested its possibility to be used as natural antioxidant. This fruit also exhibited antimicrobial activity toward Staphylococcus aureus and Escherichia coli in low doses. Finally, in vitro gastrointestinal (GI) digestion showed a partial bioaccessibility of (poly) phenols (~50% at the final step), showing high antioxidant capacity in the different GI phases. These results revealed Andean blueberry as an interesting candidate for being used as a functional ingredient and the development of further in vivo and clinical assays.

Skin Carotenoids in Public Health and Nutricosmetics: The Emerging Roles and Applications of the UV Radiation-Absorbing Colourless Carotenoids Phytoene and Phytofluene.

In this work, the importance of dietary carotenoids in skin health and appearance is comprehensively reviewed and discussed. References are made to their applications in health-promoting and nutricosmetic products and the important public health implications that can be derived. Attention is focused on the colourless UV radiation (UVR)-absorbing dietary carotenoids phytoene and phytofluene, which are attracting increased interest in food science and technology, nutrition, health and cosmetics. These compounds are major dietary carotenoids, readily bioavailable, and have been shown to be involved in several health-promoting actions, as pinpointed in recent reviews. The growing evidence that these unique UVR-absorbing carotenoids with distinctive structures, properties (light absorption, susceptibility to oxidation, rigidity, tendency to aggregation, or even fluorescence, in the case of phytofluene) and activities can be beneficial in these contexts is highlighted. Additionally, the recommendation that the levels of these carotenoids are considered in properly assessing skin carotenoid status is made.

Free carotenoids and carotenoids esters composition in Spanish orange and mandarin juices from diverse varieties.

The carotenoid profiles of citrus fruit have been well studied, but knowledge about the xanthophylls esters pattern is more limited. In this study, the native carotenoid profiles of mandarin and orange juices were studied. Xanthophylls in oranges were esterified with lauric, myristic, palmitic and stearic acids. The fatty acids esterifying ß-cryptoxanthin in mandarins were lauric, myristic, palmitic and capric acid. In oranges, the average relationships of total free carotenoids to the sum of mono-esters and di-esters were 0.82 and 1.67, respectively, and the average relationships total mono-esters to total of di-esters was 2.51. In general, the mono-ester fraction was the most significant quantitatively. In mandarins, the average relationship of total free carotenoids to the sum of mono-esters was 0.17, which is in line with the fact that ß-cryptoxanthin was mostly esterified and not free (unesterified).

Simultaneous determination of dietary isoprenoids (carotenoids, chlorophylls and tocopherols) in human faeces by Rapid Resolution Liquid Chromatography.

An analytical method was validated for the quantitative determination of isoprenoids compounds in faecal samples, based on liquid-liquid extraction from a small aliquot (0.3-0.5 g of sample) and subsequent analysis by Rapid Resolution Liquid Chromatography (RRLC) on a C30 column. An excellent linear response was observed over the range specified for all dietary isoprenoids, as confirmed by the correlation coefficient, which ranged from 0.9977 to 0.9999. LODs ranged from 0.002 µg to 0.036 µg for lutein and α-tocopherol, respectively. Depending on the compound, LOQs ranged from 0.001 µg (lutein) to 0.120 µg (α-tocopherol). For accuracy testing, spiking of faeces samples with trans-ß-apo-8'-carotenal, α-tocopherol and chlorophyll a were performed (three concentration levels). Excellent recoveries were obtained in all levels (>90%). The intra-day RSD% ranged from 0.86 to 9.78%. The inter-day RSD% was not higher than 10%, except to α-tocopherol (11.34%). In order to assess the applicability of the method faecal samples from a baby fed with different purees formulated from various vegetables were analysed during a six month period. α-carotene, ß-carotene, capsanthin, lycopene, lutein, phytoene, phytofluene, violaxanthin, zeaxanthin and ζ-carotene), and their isomers were identified and quantified using this method. Besides, 2 tocopherols and 9 chlorophylls and derivatives were identified and quantified in the faecal samples analysed. This method is suitable to determine dietary isoprenoids from complex matrices such as human faeces within 28 min.

Effect of high-pressure processing on carotenoids profile, colour, microbial and enzymatic stability of cloudy carrot juice.

The objective of this work was to assess the impact of high-pressure processing (HPP) on the carotenoid profile, colour as well as the microbial and enzymatic stability of cloudy carrot juice. The predominant carotenoids in the fresh juices were by far the provitamin A carotenoids ß-carotene and α-carotene. Others were ζ-carotene, phytofluene, phytoene and lutein. HPP at 300 MPa in three cycles caused the highest carotenoids degradation (41%) whereas the lowest degradation (26%) was achieved at 600 MPa. The highest inactivation of POD (31%) and PPO (57%) was achieved with 600 MPa and 300 MPa applied in three cycles, respectively what indicates that POD is more responsible for carotenoids degradation. The colour differences (ΔE*ab) between fresh juice and HPP-treated juices ranged from 3.02 to 4.15 CIELAB units. As far as the impact on microorganism was concerned, there was a clear trend between the applied pressure and the microbial reduction achieved.

Study of commercial quality parameters, sugars, phenolics, carotenoids and plastids in different tomato varieties.

The aim of this study was to assess commercial quality parameters, sugars, phenolics, carotenoids and plastid in diverse and little studied tomato varieties to gain insight into their commercial and functional quality and reveal possible noticeable differences. Five cherry tomato varieties and six common (i.e., non-cherry) tomatoes were evaluated. The highest levels of lycopene were detected in 'Tigerella' and 'Byelsa', and those of phytoene in 'Orange', those of phenolics in 'Green Zebra', all of them common tomatoes. The levels of sugars in both groups of tomatoes were comparable. Interesting differences in plastid carotenoid-accumulating sub-structures as a function of the carotenoid profile were observed. Given the importance of chromoplasts in the deposition of carotenoids in plants and their release during digestion, this information can be valuable in investigations on the regulation of the biosynthesis and the bioavailability of tomato carotenoids.

Comparative study of the bioaccessibility of the colorless carotenoids phytoene and phytofluene in powders and pulps of tomato: microstructural analysis and effect of addition of sunflower oil.

The objective was to assess the potential bioavailability of phytoene (PT) and phytofluene (PTF) from tomato powders used as raw materials for supplements as compared to the pulp of a common tomato and a cherry tomato. PT and PTF are attracting much interest nowadays as they can provide health and cosmetic benefits. PT and PTF levels in the more concentrated powder were up to 1000 times higher than in the tomatoes. The bioaccessibility from the powders was lower as compared to the tomato fruits and increased markedly when sunflower oil was added. However, the best source of potentially absorbable PT and PTF (0.5 and 2 mg g-1 respectively) was by far the powder with higher levels of them. This result could be due to the higher carotenoid concentration in the powder, the reduction of the particle sizes, and the rupture of cell structures compared to the pulps.

Internal preference mapping of milk-fruit beverages: Influence of color and appearance on its acceptability.

The individual preferences of 100 consumers between 20 and 30 years old for the color of 16 milk-fruit juice beverages (MFJB) were investigated by preference mapping technique. Consumers were asked to evaluate, just by looking at the samples, how much they liked them (from "Extremely dislike" to "Extremely like"). The color of the samples was analyzed by two different instrumental techniques. Results obtained from the instrumental color measurement showed the wide diversity in hues of the beverages available in the market, and correlations between techniques proved that both of them were appropriate to analyze color. Results showed that participants preferred samples with orangish appearance instead of those with a whiter look. Anyway, punctuations given by the consumers suggest that generally, color of these products is not highly evaluated by consumers, as the best mean punctuation was 6.6.