Bioavailability of Lutein from Marigold Flowers (Free vs. Ester Forms): A Randomised Cross-Over Study to Assess Serum Response and Visual Contrast Threshold in Adults.

Lutein (Lut) and zeaxanthin (Zeax) are found in the blood and are deposited in the retina (macular pigment). Both are found in the diet in free form and esterified with fatty acids. A high intake and/or status is associated with a lower risk of chronic diseases, especially eye diseases. There is a large global demand for Lut in the dietary supplement market, with marigold flowers being the main source, mainly as lutein esters. As the bioavailability of Lut from free or ester forms is controversial, our aim was to assess the bioavailability of Lut (free vs. ester) and visual contrast threshold (CT). Twenty-four healthy subjects (twelve women, twelve men), aged 20-35 and 50-65 years, were enrolled in a cross-sectional study to consume 6 mg lutein/day from marigold extract (free vs. ester) for two months. Blood samples were taken at baseline and after 15, 40, and 60 days in each period. Serum Lut and Zeax were analysed using HPLC, and dietary intake was determined with a 7-day food record at the beginning of each period. CT, with and without glare, was at 0 and 60 days at three levels of visual angle. Lut + Zeax intake at baseline was 1.9 mg/day, and serum lutein was 0.36 µmol/L. Serum lutein increased 2.4-fold on day 15 (up to 0.81 and 0.90 µmol/L with free and ester lutein, respectively) and was maintained until the end of the study. Serum Zeax increased 1.7-fold. There were no differences in serum Lut responses to free or ester lutein at any time point. CT responses to lutein supplementation (free vs. ester) were not different at any time point. CT correlated with Lut under glare conditions, and better correlations were obtained at low frequencies in the whole group due to the older group. The highest correlations occurred between CT at high frequency and with glare with serum Lut and Lut + Zeax. Only in the older group were inverse correlations found at baseline at a high frequency with L + Z and with Lut/cholesterol and at a low frequency with Lut/cholesterol. In conclusion, daily supplementation with Lut for 15 days significantly increases serum Lut in normolipemic adults to levels associated with a reduced risk of age-related eye disease regardless of the chemical form of lutein supplied. Longer supplementation, up to two months, does not significantly alter the concentration achieved but may contribute to an increase in macular pigment (a long-term marker of lutein status) and thus improve the effect on visual outcomes.

Sustainable extraction methods of carotenoids from mango (Mangifera indica L. 'Kent') pulp: Ultrasound assisted extraction and green solvents.

Mango is a good source of carotenoids for use in food, cosmetic, and pharmaceutical products because of their organoleptic and health-promoting properties. Safe and sustainable methods for their extraction is required. The present investigation was aimed to study concentration and carotenoid profile of 'Kent' mango pulp through a conventional extraction (CE) and ultrasound-assisted extraction (UAE) using traditional solvents (tetrahydrofuran-THF and diethyl ether: petroleum ether-DE:PE) and green solvents (GS) (2-metiltetrahydrofuran, 2 m-THF; cyclopentyl methyl ether, CPME). Mango showed (µg/g d.w.) ß-carotene (29.4), zeaxanthin (1.28), ß-cryptoxanthin (2.8), phytoene (18.68) and phytofluene (7.45) in a CE using DE:PE. Similar results were obtained applying DE:PE in UAE and GS in a CE, so CPME and 2-mTHF seem suitable solvents to replace DE:PE in CE. The yield of total carotenes, xanthophylls and carotenoids using GS combined with UAE was lower than with CE, but important enough to be used as a sustainable procedure for obtaining carotenoids from mango pulp.

Assessment of 24 h Sodium and Potassium Urinary Excretion in Normotensive and Hypertensive Dominican Adults.

Higher salt (sodium) intake has been associated with higher blood pressure (BP). The degree of association may be influenced by factors such as age, origin, and dietary components. This study aimed to evaluate the 24 h urinary sodium (Na) and potassium (K) excretion in normotensive and hypertensive Dominican adults and estimate their salt intake. 163 volunteers (18-80 years old) participated in a cross-sectional study. The 24 h Na and K urinary excretion were measured using an ion-selective electrode technique. Na and K urinary excretion (99.4 ± 46.5 and 35.0 ± 17.5 mmol/24 h) did not correlate with BP, except in the normotensive group, in which K correlated with SBP (0.249, p = 0.019). Na and K excretion were similar in normotensive and hypertensive subjects. When considering two age groups (18-45, 46-80 years), the Na-to-K molar ratio (3.1 ± 1.3) was higher in younger subjects (p = 0.040). Na-to-K ratio was associated with DBP in the total group (r = 0.153, p = 0.052), in the hypertensive group (r = 0.395, p < 0.001), and in the older group with SBP (0.350, p = 0.002) and DBP (0.373, p < 0.001). In the older group, Na-to-K ratio and DBP correlated after controlling for subjects with hypertension controlled by treatment (r = 0.236, p = 0.041). The Na-to-K ratio correlated, when salt intake was over 5 g/day (52.2%), with SBP (rho = 0.219, p = 0.044) and DBP (rho = 0.259, p = 0.017). Determinants of BP in the total sample were age (SBP, beta: 0.6 ± 0.1, p < 0.001; DBP, beta: 0.2 ± 0.1, p < 0.002), sex (SBP, beta: 11.2 ± 3.5, p = 0.001), body mass index (BMI) (SBP, beta: 1.0 ± 0.3, p < 0.001; DBP, beta: 0.4 ± 0.2, p = 0.01), and Na-to-K ratio (SBP, beta: 3.0 ± 1.1, p = 0.008; DBP, beta: -12.3 ± 4.0, p = 0.002). Sex and BMI were determinants in the younger group. Na-to-K molar ratio was determinant in the older group (SBP, beta: 6.7 ± 2.4, p = 0.005; DBP, beta: 3.8 ± 1.1, p < 0.001). The mean Na and salt intakes (2.3 and 5.8 g/day) were slightly higher and the K intake lower (1.4 g/day) than WHO recommendations.

Carotenoid Markers of Dietary Exposure and Nutritional Status.

Carotenoids are lipophilic isoprenoid compounds synthesized by photosynthetic organisms and some non-photosynthethic prokaryotes and fungi [...].

Natural pigments of plant origin: Classification, extraction and application in foods.

Color is the first attribute that influences the acceptance of foods as they become more attractive to consumers when they are more intense and uniform. In recent years, the interest of producers and consumers in purchasing products containing natural ingredients have constantly increased. Natural pigments are commonly extracted from fruits and vegetables, and have been proven to provide health benefits to reduce the risk of diseases such as type-1 diabetes, obesity, and coronary diseases. Additionally, advanced extraction and encapsulation technologies that make food matrices more efficient are useful tools for improving the use of these pigments. In this review, the state-of-the-art of pigments such as carotenoids, anthocyanins, and betalains of plant origin are discussed, including their main sources of production and the factors that affect their physicochemical stability. In addition, different extraction methods are discussed, listing their advantages and disadvantages, and providing some applications of natural pigments in food.

Status and Dietary Intake of Phytoene and Phytofluene in Spanish Adults and the Effect of a Four-Week Dietary Intervention with Lutein-Rich Fruits or Vegetables.

Phytoene (PT) and phytofluene (PTF) are colourless carotenoids presents in the human diet and in blood, faeces and tissues and are biologically active. However, there is very little data on these carotenoids. This study aims to assess PT and PTF concentrations in serum from healthy Spanish normolipemic subjects (n = 101, 45-65 years) and the effect of a fruit and vegetable dietary intervention (4 weeks, n = 29) on PT and PTF concentration in serum and faeces and dietary intake. Serum and faecal concentrations were analysed by HPLC and dietary intake by 3 × 24 h recalls. PT showed higher concentrations than PTF in serum, faeces and in the dietary intake. Considering both studies, PT and PTF concentrations in serum were 0.16 ± 0.07 and 0.05 ± 0.04 µmol/L, respectively, in faeces 17.7 ± 20.3 and 6.5 ± 7.9 µg/g, respectively, and in dietary intake the median was 2.4 and 0.6 mg/p/day, respectively. Carrots and tomatoes were the major dietary contributors of these carotenoids. The dietary intervention did not cause significant variations in the PT and PTF intake or serum concentrations, but a lower concentration in faeces was observed for the fruit group (PT: p = 0.024; PTF isomer-3: p = 0.034). These data highlight the need for further research on the activities of these carotenoids in humans.

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.

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.

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.

Changes in Lutein Status Markers (Serum and Faecal Concentrations, Macular Pigment) in Response to a Lutein-Rich Fruit or Vegetable (Three Pieces/Day) Dietary Intervention in Normolipemic Subjects.

Lutein is mainly supplied by dietary fruit and vegetables, and they are commonly jointly assessed in observational and interventional studies. Lutein bioavailability and health benefits depend on the food matrix. This study aimed to assess the effect of dietary intervention with lutein-rich fruit or vegetables on lutein status markers, including serum and faecal concentrations (by high pressure liquid chromatography), dietary intake (24 h recalls ×3), and macular pigment optical density (MPOD) and contrast threshold (CT) as visual outcomes. Twenty-nine healthy normolipemic subjects, aged 45-65 y, consumed 1.8 mg lutein/day supplied from fruits (14 subjects, 500 g/day of oranges, kiwi and avocados) or vegetables (15 subjects, 180 g/day of green beans, pumpkin, and sweet corn) for four weeks. Serum lutein concentration increased by 37%. The effect of the food group intervention was statistically significant for serum lutein+zeaxanthin concentration (p = 0.049). Serum α- and ß-carotene were influenced by food type (p = 0.008 and p = 0.005, respectively), but not by time. Serum lutein/HDL-cholesterol level increased by 29% (total sample, p = 0.008). Lutein+zeaxanthin/HDL-cholesterol increased, and the intervention time and food group eaten had an effect (p = 0.024 and p = 0.010, respectively) which was higher in the vegetable group. The MPOD did not show variations, nor did it correlate with CT. According to correlation matrixes, serum lutein was mainly related to lutein+zeaxanthin expressed in relation to lipids, and MPOD with the vegetable group. In faecal samples, only lutein levels increased (p = 0.012). This study shows that a relatively low amount of lutein, supplied by fruit or vegetables, can have different responses in correlated status markers, and that a longer intervention period is needed to increase the MPOD. Therefore, further study with larger sample sizes is needed on the different responses in the lutein status markers and on food types and consumption patterns in the diet, and when lutein in a "pharmacological dose" is not taken to reduce a specific risk.

Evaluation of the potential of total proanthocyanidin content in feces as an intake biomarker.

Due to the health benefits associated with proanthocyanidins (PAs), it is useful to identify dietary PA biomarkers that can be determined by simple methods. Since increased levels of circulating PA metabolites are associated with increased fecal PA content, this study explores the spectrophotometric measurement of fecal PA content and its use as a biomarker of PA intake. To this end, fecal PA content was measured using an adaptation of Porter's spectrophotometric method in samples from a preclinical study and an observational study. In the former, excretion of 250-400 mg PA polymer equivalents/100 g feces was observed during supplementation and the day after, together with a significant association (p < 0.05) between PA intake and the excretion of both intact PAs and some PA metabolites, i.e., (+)-catechin, (-)-epicatechin and syringic acid. No relationship between intake and excretion was found in the observational study, either for the entire group (mean excretion of 240 ± 226 mg PA polymer equivalents/100 g feces) or after stratification into tertiles of consumption. In conclusion, the spectrophotometric determination of total PA content in feces proved to be a valid compliance marker in a preclinical study, but it was not associated with PA intake in free-living subjects. The potential of total PA excretion in observational studies, determined in fecal samples collected the day before dietary recall or in several fecal samples from the same subject, remains to be elucidated, as does a complete validation of the method proposed here.

Predictors of macular pigment and contrast threshold in Spanish healthy normolipemic subjects (45-65 years) with habitual food intake.

INTRODUCTION: The dietary carotenoids lutein (L) and zeaxanthin (Z) are transported in the bloodstream by lipoproteins, sequestered by adipose tissue, and eventually captured in the retina where they constitute macular pigment. There are no L&Z dietary intake recommendations nor desired blood/tissue concentrations for the Spanish general population. Our aim was to assess the correlation of L&Z habitual dietary intake (excluding food supplements), resulting serum concentrations and lipid profile with macular pigment optical density (MPOD) as well as the contrast sensitivity (CT), as visual outcome in normolipemic subjects (n = 101) aged 45-65. METHODS: MPOD was measured by heterochromatic flicker photometry, serum L&Z by HPLC, the dietary intake by a 3-day food records and CT using the CGT-1000-Contrast-Glaretester at six stimulus sizes, with and without glare. RESULTS: Lutein and zeaxanthin concentrations (median) in serum: 0.361 and 0.078 µmol/L, in dietary intake: 1.1 mg L+Z/day. MPOD: 0.34du. L+Z intake correlates with their serum concentrations (rho = 0.333, p = 0.001), which in turn correlates with MPOD (rho = 0.229, p = 0.000) and with fruit and vegetable consumption (rho = 0.202, p = 0.001), but not with lutein+zeaxanthin dietary intake. MPOD correlated with CT, with and without glare (rho ranges: -0.135, 0.160 and -0.121, -0.205, respectively). MPOD predictors: serum L+Z, L+Z/HDL-cholesterol (ß-coeficient: -0.91±0.2, 95%CI: -1.3,-0.5) and HDL-cholesterol (R2 = 15.9%). CT predictors: MPOD, mainly at medium and smaller visual angles (corresponding to spatial frequencies for which sensitivity declines with age) and gender (ß-coefficients ranges: -0.95,-0.39 and -0.13,-0.39, respectively). CONCLUSION: A higher MPOD is associated with a lower ratio of L+Z/HDL-cholesterol and with a lower CT (higher contrast sensitivity). The HDL-cholesterol would also act indirectly on the CT improving the visual function.

European Database of Carotenoid Levels in Foods. Factors Affecting Carotenoid Content.

Many studies indicate that diets including carotenoid-rich foods have positive effects on human health. Some of these compounds are precursors of the essential nutrient vitamin A. The present work is aimed at implementing a database of carotenoid contents of foods available in the European market. Factors affecting carotenoid content were also discussed. Analytical data available in peer-reviewed scientific literature from 1990 to 2018 and obtained by HPLC/UHPLC were considered. The database includes foods classified according to the FoodEx2 system and will benefit compilers, nutritionists and other professionals in areas related to food and human health. The results show the importance of food characterization to ensure its intercomparability, as large variations in carotenoid levels are observed between species and among varieties/cultivars/landraces. This highlights the significance of integrating nutritional criteria into agricultural choices and of promoting biodiversity. The uncertainty quantification associated with the measurements of the carotenoid content was very rarely evaluated in the literature consulted. According to the EuroFIR data quality evaluation system for food composition tables, the total data quality index mean was 24 in 35, reflecting efforts by researchers in the analytical methods, and less resources in the sampling plan documentation.

A Randomized Study of Nutritional Supplementation in Patients with Unilateral Wet Age-Related Macular Degeneration.

The purpose of this study is evaluate the efficacy and safety of medicinal products containing the original Age-Related Eye Disease group (AREDS) formulation at doses approved in Europe (EU, control group; n = 59) with a product that adds DHA, lutein, zeaxanthin, resveratrol and hydroxytyrosol to the formula (intervention group; n = 50). This was a multicenter, randomized, observer-blinded trial conducted in patients aged 50 years or older diagnosed with unilateral exudative Age related Macular Degeneration AMD. At month 12, the intervention did not have a significant differential effect on visual acuity compared with the control group, with an estimated treatment difference in Early Treatment Diabetic Retinopathy Study (ETDRS) of -1.63 (95% CI -0.83 to 4.09; p = 0.192). The intervention exhibited a significant and, in most cases, relevant effect in terms of a reduction in some inflammatory cytokines and a greater improvement in the fatty acid profile and serum lutein and zeaxantin concentration. In patients with unilateral wet AMD, the addition of lutein, zeaxanthin, resveratrol, hydroxytyrosol and DHA to the AREDS EU recommended doses in the short-term did not have a differential effect on visual acuity compared to a standard AREDS EU formula but, in addition to improving the fatty acid profile and increasing carotenoid serum levels, may provide a beneficial effect in improving the proinflammatory and proangiogenic profile of patients with AMD.

An engineered extraplastidial pathway for carotenoid biofortification of leaves.

Carotenoids are lipophilic plastidial isoprenoids highly valued as nutrients and natural pigments. A correct balance of chlorophylls and carotenoids is required for photosynthesis and therefore highly regulated, making carotenoid enrichment of green tissues challenging. Here we show that leaf carotenoid levels can be boosted through engineering their biosynthesis outside the chloroplast. Transient expression experiments in Nicotiana benthamiana leaves indicated that high extraplastidial production of carotenoids requires an enhanced supply of their isoprenoid precursors in the cytosol, which was achieved using a deregulated form of the main rate-determining enzyme of the mevalonic acid (MVA) pathway. Constructs encoding bacterial enzymes were used to convert these MVA-derived precursors into carotenoid biosynthetic intermediates that do not normally accumulate in leaves, such as phytoene and lycopene. Cytosolic versions of these enzymes produced extraplastidial carotenoids at levels similar to those of total endogenous (i.e. chloroplast) carotenoids. Strategies to enhance the development of endomembrane structures and lipid bodies as potential extraplastidial carotenoid storage systems were not successful to further increase carotenoid contents. Phytoene was found to be more bioaccessible when accumulated outside plastids, whereas lycopene formed cytosolic crystalloids very similar to those found in the chromoplasts of ripe tomatoes. This extraplastidial production of phytoene and lycopene led to an increased antioxidant capacity of leaves. Finally, we demonstrate that our system can be adapted for the biofortification of leafy vegetables such as lettuce.

From carotenoid intake to carotenoid blood and tissue concentrations - implications for dietary intake recommendations.

There is uncertainty regarding carotenoid intake recommendations, because positive and negative health effects have been found or are correlated with carotenoid intake and tissue levels (including blood, adipose tissue, and the macula), depending on the type of study (epidemiological vs intervention), the dose (physiological vs supraphysiological) and the matrix (foods vs supplements, isolated or used in combination). All these factors, combined with interindividual response variations (eg, depending on age, sex, disease state, genetic makeup), make the relationship between carotenoid intake and their blood/tissue concentrations often unclear and highly variable. Although blood total carotenoid concentrations <1000 nmol/L have been related to increased chronic disease risk, no dietary reference intakes (DRIs) exist. Although high total plasma/serum carotenoid concentrations of up to 7500 nmol/L are achievable after supplementation, a plateauing effect for higher doses and prolonged intake is apparent. In this review and position paper, the current knowledge on carotenoids in serum/plasma and tissues and their relationship to dietary intake and health status is summarized with the aim of proposing suggestions for a "normal," safe, and desirable range of concentrations that presumably are beneficial for health. Existing recommendations are likewise evaluated and practical dietary suggestions are included.

Dietary ß-Cryptoxanthin and α-Carotene Have Greater Apparent Bioavailability Than ß-Carotene in Subjects from Countries with Different Dietary Patterns.

ß-carotene, α-carotene and ß-cryptoxanthin are greater contributors to vitamin A intake than retinol in the human diet for most people around the world. Their contribution depends on several factors, including bioavailability and capacity of conversion into retinol. There is an increasing body of research showing that the use of retinol activity equivalents or retinol equivalents could lead to the underestimation of the contribution of ß-cryptoxanthin and of α-carotene. The aim is to assess their apparent bioavailability by comparing concentrations in blood to their dietary intakes and identifying the major food contributors to their dietary intake. Dietary intake (3-day 24-h records) and serum concentrations (by HPLC) were calculated in normolipemic subjects with adequate retinol status (≥1.1 µmol/L) from our studies (n = 633) and apparent bioavailability calculated from 22 other studies (n = 29,700). Apparent bioavailability was calculated as the ratio of concentration in the blood to carotenoid intake. Apparent bioavailabilities for α-carotene and ß-cryptoxanthin were compared to those for ß-carotene. Eating comparable amounts of α-carotene, ß-cryptoxanthin and ß-carotene foods resulted in 55% greater α-carotene (95% CI 35, 90) and 686% higher ß-cryptoxanthin (95% CI 556, 1016) concentrations than ß-carotene in blood. This suggests differences in the apparent bioavailability of α-carotene and ß-cryptoxanthin and even larger differences with ß-cryptoxanthin, greater than that of ß-carotene. Four fruits (tomato, orange, tangerine, red pepper) and two vegetables (carrot, spinach) are the main contributors to their dietary intake (>50%) in Europeans.

Assessment of carotenoid concentrations in red peppers (Capsicum annuum) under domestic refrigeration for three weeks as determined by HPLC-DAD.

Red peppers (Capsicum annuum) are rich in carotenoids and are widely grown and consumed all over the world. Today's consumption patterns are characterized by periodical purchases of food and longer food storage periods, including raw fruits and vegetables, which could have a negative effect on healthy components. This study aims to investigate the individual carotenoid content in Lamuyo-variety red peppers in cool storage (7 °C) for three weeks. Carotenoid concentrations expressed in µg/100g of the edible portion were; lutein (1203), zeaxanthin (853), α-carotene (272), ß-carotene (2167), ß-cryptoxanthin (525), violaxanthin (770), capsanthin (9667), phytoene (348) and phytofluene (143). Carotenoid concentrations did not significantly vary after 21 days under household refrigeration conditions and thus the nutritional supply of provitamin A carotenoids and of carotenoids with eye health benefits such as lutein and zeaxanthin, as well as others with potential health benefits in humans such as capsanthin, violaxathin, phytoene and phytofluene.

Extraction and Analysis by HPLC-DAD of Carotenoids in Human Faeces from Spanish Adults.

Carotenoids are bioactive compounds with widely accepted health benefits. Their quantification in human faeces can be a useful non-invasive approach to assess their bioavailability. Identification and quantification of major dietary carotenoids in human faeces was the aim of the present study. Faeces and dietary intake were obtained from 101 healthy adults (45-65 years). Carotenoid concentrations were determined by HPLC in faeces and by 3-day food records in dietary intake. Carotenoids quantified in faeces (µg/g dry weight, median) were: ß-carotene (39.5), lycopene (20), lutein (17.5), phytoene (11.4), zeaxanthin (6.3), ß-cryptoxanthin (4.5), phytofluene (2.9). α-carotene (5.3) and violaxanthin were found 75.5% and 7.1% of the faeces. The carotenoids found in the highest concentrations corresponded to the ones consumed in the greatest amounts (µg/d): lycopene (13,146), phytoene (2697), ß-carotene (1812), lutein+zeaxanthin (1148). Carotenoid concentration in faeces and in dietary intake showed correlation for the total non-provitamin A carotenoids (r = 0.302; p = 0.003), phytoene (r = 0.339; p = 0.001), phytofluene (r = 0.279; p = 0.005), lycopene (0.223; p = 0.027), lutein+zeaxanthin (r = 0.291; p = 0.04) and ß-cryptoxanthin (r = 0.323; p = 0.001). A high proportion of dietary carotenoids, especially those with provitamin A activity and some of their isomers, reach the large intestine, suggesting a low bioavailability of their intact forms.

Intervention Studies in Humans.

Intervention studies in humans provide the highest level of evidence in the assessment of the effect of nutrients or bioactive food components on human health /disease. There are different intervention study designs, and their selection depends on the objectives of the study. Here we present the main designs and the factors that should be considered when selecting and developing them.