The Journal of Nutrition Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions Divalent Minerals Decrease Micellarization and Uptake of Carotenoids and Digestion Products into Caco-2 Cells 1-3

Abstract Carotenoids are lipophilic, dietary antioxidants with the potential to prevent chronic and age-related diseases. Prior to their availability for physiological functions, carotenoids require micellarization and intestinal uptake, both constituting marginally understood processes. Based on an in vitro digestion model coupled to Caco-2 cells, we assessed the effect of dietary abundant divalent ions on spinach-derived carotenoid micellarization and cellular uptake: calcium (Ca) and magnesium (Mg) ranging from 7.5 to 25 mmol/L in the digesta and zinc (Zn) and iron (Fe) ranging from 3.8 to 12.5 mmol/L. Both micellarization and uptake were significantly inhibited by minerals in a concentration-dependent manner, with stronger effects for Fe and Zn compared to Ca and Mg. Compared to controls (no mineral addition), fractional micellarization and uptake were decreased to the greatest extent (to 22.5 and 5.0%, respectively; P , 0.001) by 12.5 mmol/L Fe. Effects of Mg were of the least magnitude; at 25 mmol/L, only uptake was decreased significantly to 69.2% of the control value (P , 0.001). Total cellular carotenoid uptake from test meals decreased similarly compared to micellarization; however, decreased b-carotene micellarization was counterbalanced by improved fractional cellular uptakes from the micelles for all ions. Compared to controls, fractional b-carotene uptake from the micelles was greater in samples digested in the presence of Fe, Ca, and Zn, by up to 5-10 times at the highest concentrations of each ion (P , 0.001). Like for the above carotenoids, a high cellular uptake of the epoxycarotenoid conversion products neochrome (from neoxanthin) and luteoxanthin+auroxanthin (from violaxanthin) was also observed. The present results indicate that divalent ions may inhibit carotenoid micellarization and uptake. J. Nutr

African Adansonia digitata fruit pulp (baobab) modifies provitamin A carotenoid bioaccessibility from composite pearl millet porridges

Food-to-food fortification of staple cereal products using nutrient-dense plants shows promise to address multiple micronutrient deficiencies including vitamin A, iron and zinc in Sub-Saharan Africa. However, there is limited information on the potential interaction effects that such food-to-food fortified strategies may have on individual micronutrient bioavailability. The main objective of the current study was to investigate the impact of incorporating Adansonia digitata (baobab fruit pulp), a mineral-rich plant material, on the delivery of carotenoids from a composite cereal porridge. Formulations of native fruit/vegetable-cereal composites were screened for interactions which could influence both bioaccessibility and subsequent intestinal uptake of provitamin A carotenoids. Proportions of pearl millet flour and plant materials were dry blended to provide composite cereal porridges with total provitamin A carotenoid concentrations ranging from 3590.7 ± 23.4 to 3698.5 ± 26.5 μg/100 g (fw) and baobab concentrations ranging from 0 to 25% (dw).While there were no significant differences in provitamin A carotenoid bioaccessibility from porridge formulations containing 5 or 15% baobab, inclusion of 25% baobab resulted in a significant (p < 0.05) decrease in bioaccessibility (13.3%) as compared to the control (23.8%). Despite the reduced bioaccessibility, 6 h uptake efficiency of provitamin A carotenoids by Caco-2 human intestinal cells was not significantly altered by 25% baobab inclusion. These findings suggest that the inhibitory effects on carotenoid micellarization (bioaccessibility) observed with increased baobab addition may not ultimately limit the bioavailability of carotenoids.The USAID FoodProcessing & Post Harvest Innovation Lab (FPLAID-0AA-L-14-00003) and Sorghum & Millet Innovation Lab (SMILAID-0AA-A-13-00047) through United States Agency for International Development (USAID).http://link.springer.com/journal/131972020-11-22hj2020Consumer ScienceFood Scienc

EFECTO DE LAS CARACTERÍSTICAS QUÍMICAS Y REOLÓGICAS DE LAS PECTINAS EN LA MICELARIZACIÓN DE CAROTENOIDES

Los carotenoides son pigmentos liposolubles con efectos benéficos en la salud. Su absorción es extremadamente baja y limita su bioactividad. El proceso de absorción implica su transferencia de las gotas de grasa emulsificadas hacia las micelas. Solamente los carotenoides micelarizados pueden ser absorbidos. La micelarización de carotenoides es altamente variable, dependiendo principalmente de la matriz alimenticia que los contiene. Bajo diversas premisas se hipotetizó que la cantidad y propiedades de las pectinas contenidas en alimentos ricos en carotenoides limitan su micelarización a través de diversos mecanismos involucrados en la digestión de lípidos. Esta hipótesis se intentó demostrar tomando como modelo de estudio las pectinas y carotenoides del chile Jalapeño. Se extrajeron y caracterizaron pectinas de chile Jalapeño verde y rojo, crudo y procesado, permitiendo obtener pectinas con diversas propiedades. Se seleccionaron tres pectinas con diferente peso molecular, grado de esterificación y viscosidad. Se prepararon emulsiones aceite en agua con dos niveles (0.14% y 1%) de concentración de pectina y aceite enriquecido con carotenoides libres y esterificados de chile Jalapeño rojo. Las emulsiones se sometieron a un proceso de digestión in vitro para evaluar el impacto de la cantidad y propiedades de pectina en diversos eventos involucrados en la micelarización de carotenoides. La viscosidad del medio gastrointestinal dependió de la concentración y propiedades de las pectinas adicionadas. Dicha viscosidad fue diferente a la observada en soluciones modelo. El tamaño de partícula incrementó con la concentración de pectina en los medios intestinal y micelar. Las fibras secuestraron sales biliares, pero este efecto se rejudo al incrementar la concentración de pectina. Los resultados sugirieron que esta capacidad de secuestro dependió del grado de esterificación. La lipólisis, medida por titulación (PH-STAT), dependió de la concentración y tipo de pectina. Con altas concentraciones, la lipólisis fue más tardada. La cantidad de ácidos grasos liberados pareció estar inversamente relacionada con el peso molecular de las pectinas

Carotenoid and carotenoid esters : analysis and in vitro digestion in individual and co-consumed foods

Orientadores: Adriana Zerlotti Mercadante, Lilian Regina Barros MariuttiTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: O crescente interesse no comportamento dos carotenoides e ésteres de carotenoides durante o processo de digestão e seu impacto na saúde humana, faz dos ensaios de bioacessibilidade in vitro destes compostos um dos tópicos mais atuais de estudo na área de Alimentos e Nutrição. Aspectos desafiadores relacionados à análise de carotenoides e ésteres de carotenoides nos alimentos, requisito para determinação da sua bioacessibilidade, e aos métodos de digestão in vitro utilizados para este fim, foram abordados nesta tese. A identificação do perfil nativo de ésteres de carotenoides é frequentemente negligenciada devido à complexidade desta análise. Dentre outros fatores, interferentes lipídicos que permanecem no extrato quando a saponificação não é realizada prejudicam ou mesmo impedem a identificação dos compostos. Nesse sentido, um novo procedimento de limpeza pré-cromatográfico em duas etapas foi desenvolvido para tornar viável a análise da composição nativa de carotenoides de murici, uma fruta da Amazônia. Os interferentes (principalmente triacilglicerois) foram eficientemente removidos após separação física seguida de cromatografia em coluna aberta, possibilitando a identificação de 35 carotenoides (seis carotenoides livres, 14 monoésteres, e 15 diésteres) no extrato não saponificado de murici, por HPLC-DAD(APCI)MS/MS, enquanto apenas 6 compostos foram identificados quando a limpeza não foi realizada. Além disso, a publicação recente, pela ação INFOGEST, de um novo método de digestão in vitro de consenso internacional, chamou a atenção para a necessidade de padronização das condições de digestão simulada de alimentos em todo o mundo. O método INFOGEST, no entanto, não contempla passos chave para a determinação da bioacessibilidade de carotenoides, e tem se mostrado trabalhoso, demorado e oneroso. Diante disso, este método foi adaptado para análise de carotenoides com as etapas de separação da fração micelar e extração de carotenoides das micelas, e aplicado com sucesso na determinação da bioacessibilidade de ésteres de carotenoides de murici e de carotenoides em um amplo grupo de alimentos que são fontes destes compostos. Os resultados obtidos com o método INFOGEST adaptado foram comparados aos encontrados com métodos mais simples, tradicionalmente utilizados para estimar a bioacessibilidade de carotenoides e ésteres de carotenoides. O método INFOGEST adaptado forneceu valores de bioacessibilidade de carotenoides e ésteres de carotenoides de murici maiores do que os resultados obtidos usando o método de digestão que vinha sendo utilizado em nosso laboratório, publicado em 2014. A bioacessibilidade variou de 4 a 29%, dependendo do método utilizado e da estrutura do carotenoide, e, em geral, carotenoides livres apresentaram maior eficiência de micelarização. Quando comparado ao primeiro método de digestão in vitro adaptado para carotenoides, publicado em 1999, o método INFOGEST adaptado forneceu estimativas similares de bioacessibilidade de carotenoides tanto em alimentos individuais como combinados, e uma correlação positiva foi encontrada entre os dois conjuntos de dados. Ainda, a adição de ovo cozido à salada vegetal aumentou a bioacessibilidade in vitro de luteína e licopeno, enquanto a co-digestão com salmão promoveu maior micelarização de 'alfa'-caroteno, 'beta'-caroteno e luteína, independentemente do método de digestão empregado. Com os resultados obtidos durante este período 3 artigos foram submetidos a revisão por pares e publicados em periódicos internacionais indexadosAbstract: The interest in understanding the fate of carotenoid and carotenoid esters through the digestion process and its impact in human health is growing, making their bioaccessibility assays one of the most innovative issues of study in the field of Food and Nutrition. Challenging aspects related to the analysis of carotenoids and carotenoid esters in foods, which is a requirement for assessing their bioaccessibility, and to the in vitro digestion methods used for this purpose were addressed in this thesis. The identification of the native carotenoid profile of foods is often overlooked because of the complexity of such analysis. Among other factors, interfering lipids that remain in the extract when no saponification step is carried out impair or even preclude the compound identification. In this sense, a new pre-chromatographic two-step cleanup procedure was developed to make feasible the identification of the native carotenoid composition of murici, an Amazonian fruit. Interfering compounds (mainly triacylglycerides) were efficiently removed after physical separation followed by open column chromatography, thereby allowing the identification of 35 carotenoids (six free carotenoids, 14 monoesters and 15 diesters) in non-saponified extracts from murici by HPLC-DAD(APCI)MS/MS, whereas only 6 compounds were identified when no cleanup procedure was performed. In addition, a recent publication of a new in vitro digestion method as an international consensus by the INFOGEST action drew attention to the necessity for standardization of the in vitro digestion conditions of foods at the international level. The INFOGEST method, however, does not address crucial steps needed to assess the carotenoid bioaccessibility, and it is more laborious, time-consuming and expensive than the traditionally used ones. Therefore, the INFOGEST method was adapted for carotenoid analysis by coupling the steps of micellar fraction separation and carotenoid extraction from the micelles, which allowed the successful determination of the in vitro bioaccessibility of carotenoid esters in murici and bioaccessibility of carotenoids in a large group of carotenoid-rich foods. The results obtained with the adapted INFOGEST method were compared to those found using relatively simple and consolidated in vitro digestion models, traditionally used to estimate the bioaccessibility of carotenoids and carotenoid esters. The adapted INFOGEST method provided values of carotenoid and carotenoid ester bioaccessibility in murici higher than the results obtained using the digestion method that was being used in our laboratory, published in 2014. The carotenoid bioaccessibility ranged from 4 to 29%, depending on the digestion method and carotenoid structure, and free carotenoids overall presented higher efficiencies of micellarization than free carotenoids and monoesters. Moreover, compared with the first in vitro digestion method adapted for carotenoids, published in 1999, the adapted INFOGEST method generally provided similar estimates of carotenoid bioaccessibility during the digestion of both individual and combined foods, and a positive correlation was found between the two sets of data. Furthermore, addition of cooked egg to the vegetable salad increased the in vitro bioaccessibility of lutein and lycopene, while the co-digestion of pan-fried salmon promoted the micellarization of 'alpha'-carotene, 'beta'-carotene and lutein, regardless of the digestion method employed. Three scientific papers were published in international peer-reviewed indexed journals with the results obtained during this periodDoutoradoCiência de AlimentosDoutora em Ciência de Alimentos2013/23218-1FAPES

In Vitro Models for Studying Secondary Plant Metabolite Digestion and Bioaccessibility

There is an increased interest in secondary plant metabolites, such as polyphenols and carotenoids, due to their proposed health benefits. Much attention has focused on their bioavailability, a prerequisite for further physiological functions. As human studies are time consuming, costly, and restricted by ethical concerns, in vitro models for investigating the effects of digestion on these compounds have been developed and employed to predict their release from the food matrix, bioaccessibility, and assess changes in their profiles prior to absorption. Most typically, models simulate digestion in the oral cavity, the stomach, the small intestine, and, occasionally, the large intestine. A plethora of models have been reported, the choice mostly driven by the type of phytochemical studied, whether the purpose is screening or studying under close physiological conditions, and the availability of the model systems. Unfortunately, the diversity of model conditions has hampered the ability to compare results across different studies. For example, there is substantial variability in the time of digestion, concentrations of salts, enzymes, and bile acids used, pH, the inclusion of various digestion stages; and whether chosen conditions are static (with fixed concentrations of enzymes, bile salts, digesta, and so on) or dynamic (varying concentrations of these constituents). This review presents an overview of models that have been employed to study the digestion of both lipophilic and hydrophilic phytochemicals, comparing digestive conditions in vitro and in vivo and, finally, suggests a set of parameters for static models that resemble physiological conditions

Low β-carotene bioaccessibility and bioavailability from high fat, dairy-based meal

PURPOSE : The original aim of the study was to determine, in a double-blind 3-arm crossover human trial (n = 7), the effect of supplemental levels of iron (25 mg) and zinc (30 mg) on β-carotene (synthetic) bioavailability (10 h postprandial). However, despite the high dose of supplemental β-carotene (15 mg) consumed with the high fat (18 g), dairy-based breakfast test meal, there was a negligible postprandial response in plasma and triglyceride rich fraction β-carotene concentrations. We then systematically investigated the possible reasons for this low bioavailability of β-carotene. METHODS : We determined (1) if the supplemental β-carotene could be micellised and absorbed by epithelial cells, using a Caco-2 cell model, (2) if the fat from the test meal was sufficiently bioavailable to facilitate β-carotene bioavailability, (3) the extent to which the β-carotene could have been metabolised and converted to retinoic acid/retinol and (4) the effect of the test meal matrix on the β-carotene bioaccessibility (in vitro digestion) and Caco-2 cellular uptake. RESULTS : We found that (1) The supplemental β-carotene could be micellised and absorbed by epithelial cells, (2) the postprandial plasma triacylglycerol response was substantial (approximately 75–100 mg dL−1 over 10 h), indicating sufficient lipid bioavailability to ensure β-carotene absorption, (3) the high fat content of the meal (approximately 18 g) could have resulted in increased β-carotene metabolism, (4) β-carotene bioaccessibility from the dairy-based test meal was sixfold lower (p < 0.05) than when digested with olive oil. CONCLUSION : The low β-carotene bioavailability is probably due to a combination of the metabolism of β-carotene to retinol by BCMO1 and interactions of β-carotene with the food matrix, decreasing the bioaccessibility. TRAIL REGISTRATION : The human trail was retrospectively registered (ClinicalTrail.gov ID: NCT05840848).A Georg Forster Research Fellowship from the Alexander von Humboldt Foundation. Open Access funding enabled and organized by Projekt DEAL.http://link.springer.com/journal/394hj2024Consumer ScienceFood ScienceSDG-02:Zero Hunge

Improving the Bioaccessibility and Bioavailability of Carotenoids by Means of Nanostructured Delivery Systems: A Comprehensive Review

Carotenoids are bioactive compounds provided by the diet playing a key role in maintaining human health. Therefore, they should be ingested daily in an adequate amount. However, even a varied and well-balanced diet does not guarantee an adequate intake, as both the bioaccessibility and bioavailability of the compounds significantly affect their absorption. This review summarizes the main results achieved in improving the bioaccessibility and bioavailability of carotenoids by means of nanostructured delivery systems, discussing in detail the available lipid-based and biopolymeric nanocarriers at present, with a focus on their formulation and functional efficiency. Although the toxicity profile of these innovative delivery systems is not fully understood, especially for long-term intake, these systems are an effective and valuable approach to increase the availability of compounds of nutritional interest

Improving the Bioaccessibility and Bioavailability of Carotenoids by Means of Nanostructured Delivery Systems: A Comprehensive Review

Carotenoids are bioactive compounds provided by the diet playing a key role in maintaining human health. Therefore, they should be ingested daily in an adequate amount. However, even a varied and well-balanced diet does not guarantee an adequate intake, as both the bioaccessibility and bioavailability of the compounds significantly affect their absorption. This review summarizes the main results achieved in improving the bioaccessibility and bioavailability of carotenoids by means of nanostructured delivery systems, discussing in detail the available lipid-based and biopolymeric nanocarriers at present, with a focus on their formulation and functional efficiency. Although the toxicity profile of these innovative delivery systems is not fully understood, especially for long-term intake, these systems are an effective and valuable approach to increase the availability of compounds of nutritional interest

Quality and Nutritional Impacts of Extrution on Pearl Millet and Nutrient Dense Native Plant Blends

Significant efforts have been placed on the development of strategies to address micronutrient deficiencies in Sub-Saharan parts of Africa. Improving the nutritional value of staple foods, including cereals, is a key component of such strategies as traditional cereals such as millet and sorghum are mostly consumed in these areas. Of micronutrient deficiencies, iron, zinc and vitamin A are most critical, and mostly deficient in staple cereal foods. One option to improve the nutrient density of cereal foods is to leverage natural micronutrient dense plant materials that exist locally, and can be formulated into cereals products. Such products can be combined with processing strategies to produce a diverse array of locally produced consumer products that can be adopted by at risk populations. Success of such strategies requires significant insights into the compatibility of traditional cereal and local plant products with processing strategies. This includes new and novel drying and cereal processing strategies currently being implemented in Africa, such as solar drying and extrusion. With this in mind, the objective of the current studies was to address both the development of knowledge related to the impact of a novel solar drying system on micronutrient rich plants. The same objective was oriented toward establishing insights into the impact of extrusion conditions on product quality and the stability and bioaccessibility of provitamin A carotenoids from blended millet instant thin and thick porridges. The initial studies focused on assessing a novel solar drying system as a mode to generate high quality nutrient dense plant ingredients. Our specific focus was on stability of provitamin A carotenoids from carrot and mango fruit, two products commonly available in Africa and that suffer from high post-harvest losses. With final moistures between 7 to 11%, solar dried sliced mango showed dark off color formation as a result of extended drying times compared to carrot. Grated carrot samples, showed lower content of provitamin A carotenoids (p\u3c0.05) when subjected to extended drying as compared to sliced carrot sample. However, grated carrot samples had higher content of provitamin A carotenoids than sliced carrot samples compared to standard drying (thin layer electric and dehydrator) methods suggesting the suitability of solar drying for the development of carrot and mango powders to be used as ingredients in processed foods targeting delivery of provitamin A carotenoids. The second and third studies assessed the impact of micronutrient rich plant ingredients selected based on their content of iron, and vitamin A on quality of extruded cereal products. Formulation of blends were accomplished with whole grain (WG) millet (80%), dried carrot (Carrot-15%), Adansiona digitata (Baobab-5%) or Moringa oleifera (Moringa-5%). Blends were extruded on a Technochem mini-extruder currently being commercialized in rural and urban markets in Africa. As fully gelatinized flours, water absorption indexes were significantly increased with addition of Carrot and Baobab. Addition of Baobab and Moringa decreased product final viscosities, storage (G\u27) and loss modulus (G ). Pooling results from all added plant materials in extruded WG millet, Moringa exhibited high enthalpy and Cinfinity or C ∞ (45.7 %) values, however, starch digestion was not significantly impacted by presence of these ingredients. Regarding provitamin A carotenoid stability, high recoveries (69–90%) were obtained with inclusion of Baobab and Moringa compared to WG millet Carrot blends, (~60% recovery). Results suggest that production of naturally fortified millet blends can be achieved without significant compromise to product quality and recovery of provitamin A carotenoids. The final study further examined the impact of extrusion on the bioaccessibility of provitamin A carotenoids and chemical changes from blends of millet and plant. Generally, water solubility index was significantly increased by inclusion of CRT and Baobab (P\u3c0.05), while starch digestibility was not altered by formulation. In comparing micronutrient delivery, higher relative provitamin A bioaccessibility (~20%) and higher absolute provitamin A bioaccessibility (707.13 µg/100g porridge) were obtained from extruded WG millet, carrot, and Baobab (WG/CRT/Baobab) compared to others, suggesting potential stabilization of carotenoids by Baobab and the possibility of improving bioavailability through formulation and processing of cereal blends. Taken together, these results suggested that low cost extrusion of WG millet combined with appropriate local nutrient dense plant ingredients generated by solar drying could be used to deliver provitamin A carotenoids to at risk populations. Without altering starch digestibility significantly, extruded blends of WG millet with carotenoid sources and Baobab have potential to stabilize and deliver products with improved provitamin A bioaccessibility. Further applied research focused on solar drying tests in in Africa as well as generation and assessment of consumer product acceptability and efficacy are needed to expand upon these initial finding