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      An in vitro digestion method adapted for carotenoids and carotenoid esters: moving forward towards standardization

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          Abstract

          The consensus INFOGEST in vitro digestion method was adapted for carotenoids. The systematic identification of carotenoid esters before and after in vitro digestion is presented for the first time.

          Abstract

          In vitro digestion methods are a useful approach to predict the bioaccessibility of food components and overcome some limitations or disadvantages associated with in vivo methodologies. Recently, the INFOGEST network published a static method of in vitro digestion with a proposal for assay standardization. The INFOGEST method is not specific for any food component; therefore, we aimed to adapt this method to assess the in vitro bioaccessibility of carotenoids and carotenoid esters in a model fruit ( Byrsonima crassifolia). Two additional steps were coupled to the in vitro digestion procedure, centrifugation at 20 000 g for the separation of the aqueous phase containing mixed micelles and exhaustive carotenoid extraction with an organic solvent. The effect of electrolytes, enzymes and bile acids on carotenoid micellarization and stability was also tested. The results were compared with those found with a simpler method that has already been used for carotenoid bioaccessibility analysis. These values were in the expected range for free carotenoids (5–29%), monoesters (9–26%) and diesters (4–28%). In general, the in vitro bioaccessibility of carotenoids assessed by the adapted INFOGEST method was significantly higher ( p < 0.05) than those assessed by the simplest protocol, with or without the addition of simulated fluids. Although no trend was observed, differences in bioaccessibility values depended on the carotenoid form (free, monoester or diester), isomerization ( Z/ E) and the in vitro digestion protocol. To the best of our knowledge, it was the first time that a systematic identification of carotenoid esters by HPLC-DAD-MS/MS after in vitro digestion using the INFOGEST protocol was carried out.

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          Bioaccessibility of carotenoids and vitamin E from their main dietary sources.

          Victor Pirisi, Patrick Borel, Myriam Richelle … (2006)
          Vitamin E and carotenoids are fat-soluble microconstituents that may exert beneficial effects in humans, including protection against cancer, cardiovascular diseases, and age-related eye diseases. Their bioavailability is influenced by various factors including food matrix, formulation, and food processing. Since human studies are labor-intensive, time-consuming, and expensive, the in vitro model used in this study is increasingly being used to estimate bioaccessibility of these microconstituents. However, the ability of this model to predict bioavailability in a healthy human population has not yet been verified. The first aim of this study was to validate this model by comparing model-derived bioaccessibility data with (i) human-derived bioaccessibility data and (ii) published mean bioavailability data reported in studies involving healthy humans. The second aim was to use it to measure alpha- and gamma-tocopherol, beta-carotene, lycopene, and lutein bioaccessibility from their main dietary sources. Bioaccessibility as assessed with the in vitro model was well correlated with human-derived bioaccessibility values (r = 0.90, p beta-carotene and alpha-carotene > lycopene and alpha-tocopherol generally > gamma-tocopherol), (ii) food matrix, and (iii) food processing.
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            In VitroModels for Studying Secondary Plant Metabolite Digestion and Bioaccessibility

            T Requena, G.J. McDougall, A. Gomes … (2014)
            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.
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              Development of an in vitro digestion method to assess carotenoid bioavailability from meals.

              Laura Garrett, J Sarama, Valérie Failla (1999)
              The objective of this study was to develop a model for assessing the bioavailability of carotenoids from meals using an in vitro digestion procedure. A meal was prepared using baby food carrots, spinach, and a meat, plus tomato paste. The aqueous fraction was isolated from digesta to determine the quantity of carotenoids transferred from the food to micelles. The micellarization of lutein (25-40%) exceeded (p < 0.01) that of alpha- and beta-carotene (12-18%) and lycopene (<0.5%). Micellarization of carotenoids was not affected by elimination of the gastric phase of the digestive process. The absence of bile extract prevented the transfer of carotenoids from foods to micelles, whereas omission of pancreatin only reduced the micellarization of the carotenes. Differentiated cultures of Caco-2 human intestinal cells accumulated 28-46% of micellarized carotenoids from the medium after 6 h. These results support the usefulness of the in vitro digestion process as a rapid and cost-effective model for screening the bioavailability of carotenoids from meals.
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                Author and article information

                Journal
                Journal ID (publisher-id): FFOUAI
                Title: Food & Function
                Abbreviated Title: Food Funct.
                Publisher: Royal Society of Chemistry (RSC)
                ISSN (Print): 2042-6496
                ISSN (Electronic): 2042-650X
                Publication date Created: 2016
                Publication date (Electronic): 2016
                Volume: 7
                Issue: 12
                Pages: 4992-5001
                Affiliations
                [1 ]Department of Food Science
                [2 ]Faculty of Food Engineering
                [3 ]University of Campinas
                [4 ]Brazil
                Article
                DOI: 10.1039/C6FO01293K
                SO-VID: 504b5586-c312-4595-bbb2-837080706ef1
                Copyright © © 2016
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