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Manipulation of Starch Digestibility in Particle FormDobson, Corrine 31 October 2019 (has links)
This work investigates ways to prevent and manage hyperglycemia using preventive nutrition. Uncontrolled and chronic hyperglycemia is a global health issue leading to many health problems including diabetes. This thesis details the manipulation of highly retrograded starch particles in order to produce particles that are digested slowly to release glucose at a prolonged and moderate rate to prevent this. The first section of this study utilized acid hydrolysis to alter starch structure and change digestibility. The hydrolysis treatment showed that hydrolysis of native starch prior to particle formation changed the structure in a way that increased digestibility. The second section of this work introduced polyphenols into the particles which only a marginal effect on digestion. Overall the actual process of retrograding and making the particles themselves appeared to create particles that were more resistant to digestion. These could be used in a product to deliver a moderate glycemic response.
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Development and application of an in vitro physicochemical upper gastrointestinal system (IPUGS) simulating the human digestive processesYoo, Ji Yeon January 2009 (has links)
Increased and combined knowledge of food processing, molecular biology, health and nutrition has triggered production of many different types of functional foods and pharmaceutics recently. The efficacy and safety of such products are being assessed prior to marketing by in vivo and/or in vitro studies. Traditional in vivo studies require excessive time, cost and labour, as well as ethical approvals with subject to humans or animals in some instances. Therefore excessive number of runs may be avoided if reliable in vitro system is available. During the course of this study, an in vitro physicochemical upper gastrointestinal tract system (IPUGS), the first of its kind in literature, has been developed to simulate the relevant conditions of the gastrointestinal tract (GIT) as closely as possible to the human physiology with multi-disciplinary approach, combining biology, physiology, gastroenterology, process technology, chemical engineering and automation. The IPUGS is aimed at having a high predictive capability towards the real digestion processes occurring in the human upper GIT which allows for examining of the bioavailability of nutrients and drugs, drug-nutrient interactions, viability of probiotics and case studies of gastrointestinal disorders. Digestion of rice and baby foods have been studied with the IPUGS by UV-spectrophotometer, HPLC, light microscope and pH meter under the conditions of normal state and common gastric disorders, such as gastroparesis, dumping syndrome, Zollinger-Ellison syndrome and hypochlorhydria. By comparing the data from many physiological and clinical sources in the literature, it would seem that the IPUGS was able to generate more reliable data compared to the existing in vitro digestion (mechanical) models in the literature. In future, computer-controlled and computer-recorded data by possibly designing a new software or equations would be desirable to implicate a better understanding of the digestive processes.
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Carotenoid In Planta Development, Storage, and Bioaccessibility: A Comprehensive Approach to Nutrient AnalysisJeffery, Jennifer L. 14 January 2010 (has links)
Plants contain a host of secondary metabolites that may be of dietary use to man. A comprehensive approach to plant-based nutrition would include investigating all aspects of a nutrient, from creation through storage and consumption. Here, experiments address each of these facets for a group of important antioxidant and pigment compounds, the carotenoids.
The carotenoid biosynthetic pathway regulatory mechanisms leading to lycopene accumulation are well defined in the model fruit, tomato. Those leading to accumulation of other carotenoids and flesh colors, however, are poorly understood. The variety of flesh colors available in watermelon fruit (red, orange, salmon yellow, and canary yellow) makes it an ideal candidate for investigating the regulation of the full pathway. Carotenoid accumulation was measured in ten watermelon varieties, representing the four flesh colors and three ploidy levels, throughout fruit maturation. It was found that the putative regulatory mechanisms controlling lycopene accumulation in red-fleshed fruit may be applied in a generalized fashion to each flesh color in respect to the major carotenoid accumulated at maturity. Additionally, triploid varieties were generally found to have higher accumulation levels than diploids, and tetraploids were intermediate to both. In addition to total carotenoid content, many factors are important in determining perceived benefit. Several of these factors involve components of the food matrix, cellular and subcellular species-specific characteristics of the food which act as barriers to nutrient release. Cell size, cell wall, and chromoplast (the carotenoid storage organelle) characteristics were observed in nine fruits and vegetables using light and transmission electron microscopy. Watermelon, tomato, and melon have the largest cells. Sweet potato, butternut squash, carrot, and mango have the most fibrous cell walls; mango and papaya additionally had the thickest walls. Chromoplast globular, tubular, crystalline, and membranous substructures were described for each food. These food matrix factors may be related to differences in carotenoid bioaccessibility between food sources. An in vitro digestion experiment was used to determine carotenoid bioaccessibility for each of these foods. Per serving, grapefruit yielded the most lycopene while carrot gave the most ?-carotene, ?-carotene, lutein, and phytoene, and mango proved a good source of violaxanthin.
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Evaluation of Post Harvest Technologies for Improving Strawberry Fruit QualityMisran, Azizah 17 May 2013 (has links)
Fragaria ananassa, generally known as strawberry is a nutritious fruit that is rich in polyphenols and widely consumed as part of a healthy diet. Anthocyanins have been found to be the main group of phenolic components present in strawberry with pelargonidin-3-glucoside as the major component in all strawberry cultivars studied. The main volatile component has been identified as caryophyllene oxide, a sesquiterpene compound that was present abundantly in all cultivars. The effect of preharvest spray applications of a formulation containing hexanal which is known to enhance membrane preservation, was evaluated using two strawberry cultivars, ‘Jewel’ and ‘Mira’. Our results suggest that preharvest spray application of hexanal formulation results in biochemical changes that alter the profile of phenolic compounds and the volatiles of the fruit.
An osmotic infusion treatment was implemented to generate an intermediate moisture food product with high quality and enhanced storage life that can be potentially used in processed food. Infusing strawberry with other functional ingredients such as fructoligosaccharides, lecithin, and ascorbic acid showed that the infused strawberry produced by this technique was structurally and organoleptically similar to fresh strawberry, and visually similar to the intial fruits. Drying of infused fruits could extend their shelf life up to several months, while providing fruit products with superior nutritional qualities.
Subjecting strawberry fruit extract to a simulated in vitro digestion of strawberry greatly affected the polyphenol composition and concentrations during different stages of digestion. The concentrations of most polyphenols increased during gastric digestion suggesting that the acidic environment of the stomach may help to release polyphenols that are bound to the strawberry matrix. Following incubation of the IN and the OUT fractions with colonic bacteria, the presence of urolithin B glucunoride in the IN fraction and the increase of ellagic acid deoxyhexoside in both the IN and OUT fractions after fermentation process, suggest that the polyphenols are catabolized into simple phenolic compounds in the colon. The remaining polyphenols as well as the catabolites in the gut, could potentially have a beneficial effect in enhancing colonial health. / Ministry of Higher Education Malaysia, Universiti Putra Malaysia
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Encapsulation of Curcumin in O/w Nanoemulsions and Its Bioaccessibility After In Vitro DigestionAhmed, Kashif 01 January 2010 (has links) (PDF)
The functional ingredient curcumin has a variety of biological and pharmacological actions, such as anti-tumor, anti-inflammatory, anti-virus, anti-oxidant, and anti-HIV properties coupled with low toxicity. However, curcumin possesses low bioavailability due to its poor solubility in water. The purpose of this study was to investigate the impact of different lipid-based formulations of curcumin on in vitro solubilization and bioaccessibility. Oils representing LCT, MCT, LCT:SCT mix and SCT were used to prepare O/W (nano)emulsions with droplet sizes as low as 174 nm. An in vitro digestion model simulating the small intestine milieu in the fasted and fed state was used to characterize rate, extent, and particle size associated with emulsion digestion. Rate and extent were oil dependent, but not particle size. SCT emulsions digested at the fastest initial rate, but MCT emulsions were digested to the largest extent. Bioaccessibility, a precursor to eventual bioavailability, was determined after digestion using a curcumin:lipid content dependent and independent method. MCT produced the highest bioaccessibility of curcumin for each method. Nanoemulsion digestion and bioaccessibility results were compared to conventional emulsions because an appropriate comparison was needed to determine the merits of the nanoemulsion delivery system. There was no significant difference in particle size and bioaccessibility between the conventional and nanoemulsions.
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Impact of fatty acyl composition and quantity of triglycerides on bioaccessibility of dietary carotenoidsHuo, Tianyao 10 December 2007 (has links)
No description available.
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Bioacessibilidade, atividade antioxidante e antiproliferativa de compostos bioativos fenólicos de sucos de frutos da família Myrtaceae / Bioaccessibility, antioxidant and antiproliferative activity of bioactive phenolic compounds in juices from fruits of Myrtaceae familyBeteto, Flávia Maria 16 November 2015 (has links)
Diversos estudos com compostos fenólicos têm demonstrado os efeitos benéficos destas substâncias frente a diversas patologias, incluindo alguns tipos de câncer. Considerando que os polifenóis da dieta, não absorvidos, podem permanecer no trato gastrointestinal por um período prolongado, e as células do epitélio intestinal podem ser regularmente expostas a estes compostos, é importante avaliar seu potencial efeito benéfico no trato gastrointestinal. Entretanto, é necessário determinar como o processo de digestão afeta a estabilidade e propriedades químicas destes compostos. O objetivo deste estudo foi avaliar a bioacessibilidade dos polifenóis de sucos de frutas da família Myrtaceae: cagaita (Eugenia dysenterica DC), camu-camu (Myrciaria dubia Mc Vaugh) e jaboticaba (Myrciaria cauliflora B.), o efeito da digestão gastrintestinal in vitro sobre sua atividade antioxidante, e a ação dos polifenóis dos sucos digeridos sobre a proliferação, ciclo celular e apoptose em células Caco-2 de adenocarcinoma de cólon humano. A digestão simulada in vitro causou perdas de alguns compostos, tais como os derivados de cianidina encontrados na jaboticaba, possivelmente devido às condições do pH intestinal. No entanto, o conteúdo de ácido elágico livre aumentou em todos os sucos analisados, indicando a ocorrência de hidrólise durante o processo de digestão in vitro, liberando ácido elágico a partir dos elagitaninos. A atividade antioxidante dos polifenóis foi afetada de forma diferente pela digestão in vitro, de acordo com o suco, provavelmente relacionado à composição de polifenóis. Quanto à proliferação, ciclo celular e apoptose, os polifenóis a partir da fração bioacessível do camu-camu apresentou aproximadamente 30% de inibição da proliferação, seguido pela cagaita com 24%, ambos na maior concentração testada (50 µg EAG/mL). Jaboticaba não apresentou efeito inibitório nas concentrações testadas, entretanto os compostos fenólicos de todas as frações bioacessíveis (50 µg EAG/mL) apresentaram parada no ciclo celular na fase G2/M sem induzir apoptose nas células Caco-2. Os resultados sugerem que os polifenóis das Myrtaceae podem modular a proliferação nas células Caco-2 por bloqueio da progressão do ciclo celular na fase G2/M e assim oferecer efeitos benéficos para a saúde do trato gastrointestinal. / Several studies with phenolic compounds have shown the beneficial effects of these substances across various diseases, including some types of cancer. Considering that most of the polyphenols and their conjugates, unabsorbed, can remain in the lumen for a prolonged period, and epithelial cells lining the intestine are regularly exposed to these compounds, it is important to evaluate their potential beneficial effects in the gastrointestinal tract. However it is necessary to evaluate how the digestion process affects the stability and chemical properties of these compounds. The aims of this study were to evaluate the bioaccessibility of polyphenols in juices from Brazilian native fruits of the Myrtaceae family (cagaita, camu-camu and jaboticaba), the effect of in vitro gastrointestinal digestion on their antioxidant activity, and the action of polyphenols from digested juices on proliferation, cell cycle and apoptosis in human colon cancer Caco-2 cells. The results showed that in vitro gastrointestinal digestion caused losses of some polyphenols, such as cyanidins derivatives from jaboticaba, possibly due to the exposure to conditions of intestinal pH. However, contents of free ellagic acid increased in all the juices analyzed, indicating the occurrence of hydrolysis during in vitro digestion process, releasing ellagic acid from the ellagitannins. The antioxidant activity was affected for different forms by the in vitro digestion, demonstrating be related to individual components present in each sample and the mechanisms by which they act as antioxidants. Regarding the evaluation of proliferation, cell-cycle and apoptosis, polyphenols from bioaccessible fractions of camu-camu showed about 30% of inhibition of proliferation, followed by cagaita with 24%, both at the highest concentration tested (50 µg GAE/mL). Jaboticaba did not show inhibitory effect at the concentrations tested but the phenolic compounds of all bioaccessible fractions (50 µg GAE/mL) showed arrest in G2/M phase of cell-cycle without inducing apoptosis in the Caco-2 cells. Results suggest that Myrtaceae polyphenols may modulate the proliferation of Caco-2 cells by blocking the progression of cell-cycle at G2/M phase, providing beneficial effects to gastrointestinal health.
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Caracterização química de tomate roxo e estudo da metabolização de flavonoides in vitro e in vivo / Chemical characterization of purple tomato and in vitro and in vivo study of metabolism of flavonoidsSouza, Mayara Adja da Silva 16 February 2018 (has links)
Tendo em vista a importância dos compostos bioativos (CBAs) para a promoção da saúde, foram desenvolvidos, a partir de cruzamentos do tomate cereja com espécies selvagens, os tomates laranja (rico em β-caroteno) e roxo (rico em antocianinas), por meio da técnica de introgressão de alelos. Diante disso, o objetivo deste trabalho foi caracterizar o perfil de compostos bioativos e voláteis dos tomates enriquecidos e avaliar a estabilidade e metabolização de flavonoides do tomate roxo durante digestão in vitro e em modelo animal. Os tomates foram caracterizados quanto ao conteúdo de compostos fenólicos totais; capacidade antioxidante por DPPH e ORAC; ácidos orgânicos; açúcares solúveis; perfil de carotenoides por CLAE/DAD; flavonoides por CLAE/DAD e LC/ESI/MS/MS e compostos voláteis por CG/MS. Avaliou-se ainda a estabilidade dos flavonoides da casca do tomate roxo por simulação da digestão in vitro, utilizando o Simulator of the Human Intestinal Microbial Ecosystem (SHIME), bem como a formação de AGCC por GC-MS, e excreção em ratos Wistar, com posterior identificação dos compostos fenólicos por LC/Q-TOF/MS. O tomate roxo apresentou aumento no conteúdo de fenólicos totais, capacidade antioxidante e vitamina C, com destaque para casca. A rutina foi o principal flavonol identificado em todos os frutos, e na casca do tomate roxo foi encontrado alto teor de petunidina (p-coumaroil)-rutinosídeo-hexosídeo, além da superexpressão de outros flavonoides como a quercetina-3-O-rutinosídeo e kaempferol. Não houve alteração no perfil de flavonoides do fruto laranja. Este, por sua vez, apresentou acúmulo de β-caroteno, importante pró-vitamina A, ao passo que o tomate roxo também teve seus conteúdos de β-caroteno e licopeno aumentados. Os frutos apresentaram perfil de compostos voláteis diferentes entre si, o que foi relacionado à degradação dos diferentes CBAs característicos de cada um. O extrato fenólico da casca de tomate roxo, submetido à digestão in vitro, se manteve estável na primeira porção, relativo às condições estomacais. Contudo, o conteúdo de flavonoides apresentou redução significativa (p<0,05) nas porções que simulam as condições do duodeno e do colón, com a formação de catabólitos pela ação da microbiota intestinal e/ou pela degradação química espontânea. Foi observado o aparecimento de novos ácidos fenólicos não presentes inicialmente na matriz, dentre eles o ácido 3-O-metilgálico e o ácido homovanílico, supostamente derivados da degradação da petunidina e da quercetina, respectivamente. Houve aumento na produção total de AGCC, com excessão do butirato. Na urina dos animais foram detectados diversos outros compostos fenólicos derivados do metabolismo de fase II, dentre eles o ácido hipúrico e o 3-O-metilcatecol. Nas fezes foram identificados cerca de metade dos compostos presentes na fermentação in vitro. Dessa forma, o melhoramento convencional pode ser uma alternativa para o enriquecimento, com CBAs, de alimentos amplamente consumidos pela população, como o tomate. Além disso, durante a passagem pelo trato gastrointestinal, os flavonoides presentes na casca do tomate roxo sofrem intensa degradação pela microbiota intestinal, com formação de catabólitos com reconhecido potencial benefício à saúde. / Considering the importance of bioactive compounds (BACs) for health promotion, the orange (β-carotene-rich) and purple (anthocyanin-rich) tomatoes were developed from cherry tomato interspecific crossing with wild species, using the technique of allele introgression. The objective of this work was to characterize the profile of bioactive and volatile compounds of enriched tomatoes, and to evaluate the stability and metabolism of purple tomato\'s flavonoids during in vitro and in vivo digestion. The tomatoes were characterized by its content of total phenolic compounds; antioxidant capacity (DPPH and ORAC); organic acids; soluble sugars; carotenoids (HPLC/DAD); flavonoids (HPLC/DAD and LC/ESI/MS/MS) and volatile compounds (GC/MS). The stability of the flavonoids from purple tomato peel was assessed by using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME), as well as the formation of AGCC by GC-MS, and metabolism and excretion in Wistar rats, with subsequent identification of phenolic compounds by LC/Q-TOF/MS. The purple tomato showed an increase in the total phenolic content, antioxidant capacity and vitamin C, with highlight for the peel. The rutin was the main flavonol identified in all fruits, and it was found high content of petunidin (p-coumaryl)-rutinoside-hexoside in the purple tomato peel, in addition to overexpression of other flavonoids such as quercetin-3-O-rutinoside and kaempferol. There was no change in the flavonoid profile of the orange fruit. This one, in turn, presented accumulation of β-carotene, important pro-vitamin A, while the purple tomato also showed an increase in its β-carotene and lycopene contents. The fruits presented different volatile compounds profile among them, which was related to the degradation of the different BACs composition of each one. In the in vitro digestion, the phenolic extract of the purple tomato peel remained stable in the first portion, relative to the stomach conditions. However, the content of flavonoids presented a significant reduction (p<0.05) in the portions simulating the duodenum and colon, with the formation of catabolites by the action of intestinal microbiota and/or spontaneous chemical degradation. It was observed the appearance of new phenolic acids that was not initially present in the matrix, among them 3-O-methylgalic acid and homovanilic acid, supposedly derived from the degradation of petunidin and quercetin, respectively. There was an increase in the total production of SCFAs, with the exception of butyrate. In the urine of the animals several other phenolic compounds derived from phase II metabolism were detected, among them hippuric acid and 3-O-methylcatechol. About half of the compounds present in the in vitro fermentation were identified in the feces. Conventional breeding may be an alternative for the enrichment, with BACs, of foods that are widely consumed by the population, such as tomatoes. In addition, during the passage through the gastrointestinal tract, the flavonoids present in the purple tomato peel are severely degraded by the intestinal microbiota, with formation of catabolites with recognized potential health benefit.
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Adapta??o de metodologia de digest?o in vitro e determina??o da bioacessibilidade in vitro de Beta -caroteno em tr?s variedades de batata-doce de polpa alaranjada. / Adaptation of in vitro digestion methodology and determination of in vitro Beta-carotene bioacessibility of three orange sweet potato varieties.Giori, Fernanda Peixoto 23 February 2010 (has links)
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Previous issue date: 2010-02-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior-CAPES / Brazil was adverted by the World Health Organization (WHO) as a sub-clinical area of
serious vitamin A deficiency. Thus, the Brazilian Agricultural Research Corporation
(EMBRAPA) is selecting and improving varieties of sweet potatoes with higher levels
of Beta-carotene, pro-vitamin A. The carotenoids provitamin A amount of in foods does
not necessarily correspond the amount that is absorbed and metabolized by the body.
For a better determination of these values and knowledge of the mechanisms of its
transport and absorption, it is necessary to understand the factors that lead to the food
matrix release, until the absorption and the influence on the promotion and maintenance
in human health. In order to perform preliminary studies of its absorption, this study
aims to determine the efficiency of micellization of Beta -carotene in orange sweet potato
(Ipomoea batatas, Lam), by applying a in vitro digestion as a tool for determining the
bioaccessibility, which is the first step for bioavailability determination. This approach
aims to simulate the oral, gastric and intestinal stages of human. The digestion was
performed with 10 g of fresh samples, homozeneided with 5% (w/w) of canola oil. The
extraction was performed with acetone and petroleum ether and the micellar fraction
with petroleum ether, NaCl 10% (w / v) and NaSO4 2% (w/v) and involves the use of
enzymes as -amylase , pepsin, bile, pancreatin, lipase and mucin, and inorganic
compounds such as KCl, KSCN, NaH2PO4, Na3PO4, NaOH, NaCl, CaCl2, HCl,
NaHCO3. The physiological variations are reproduced by the heating bath shaker with
orbital gyrus (37?C) and centrifugation (5000g, 45 min). Quantification and
determination of the profile of carotenoids were performed by high performance liquid
chromatography (HPLC) with YCM ? C30 Carotenoid S-3 4.6 x 250mm column.
Quantification of total carotenoids was performed by UV-VIs. The whole procedure
was performed under controlled temperature (25 ? C) and light. The Beta -carotene was
present mainly with levels of 86%, 73% and 82% for access 1, 2 and 3 and after
digestion, the profile of Beta -carotene has set levels of 96%, 89% and 100%, respectively.
The efficiency of micellization was 23.8%, 28% and 25% for 1.2 and 3 hits, indicating
Beta -carotene transfer of the food matrix to micelles, corresponding to bioaccessibility of
the compound. This methodology proved to be faster and cheaper, since the in vivo
studies are costly, complex and require more time. / O Brasil foi classificado pela Organiza??o Mundial da Sa?de (OMS) como ?rea de
car?ncia sub-cl?nica grave de vitamina A. Assim, a Empresa Brasileira de Pesquisa
Agropecu?ria (EMBRAPA) v?m selecionando e melhorando variedades de batata-doce
com teores maiores de Beta-caroteno, composto pr?-vitamina A. A quantidade de
caroten?ides pr?-vitamina A presentes nos alimentos n?o corresponde necessariamente
?quela quantidade absorvida e metabolizada pelo organismo. Para uma melhor
determina??o destes valores e conhecimento dos mecanismos de transporte e absor??o
deste composto, faz-se necess?rio, o entendimento dos fatores que levam ? sua libera??o
da matriz do alimento, at? a extens?o de sua absor??o, bem como a influ?ncia na
promo??o e manuten??o da sa?de humana. A fim de realizar estudos preliminares de
sua absor??o, este trabalho visa determinar a efici?ncia de miceliza??o de Beta-caroteno de
batata-doce de polpa alaranjada (Ipomoea batatas, Lam.), atrav?s da aplica??o de
digest?o in vitro, como ferramenta de determina??o da bioacessibilidade, etapa
preliminar para a determina??o da biodisponibilidade. Esta metodologia visa simular as
etapas de digest?o oral, g?strica e intestinal humana. Foram pesados 10g de amostra in
natura e adicionados 5% (p/p) de ?leo de canola. A extra??o do alimento foi realizada
com acetona e ?ter de petr?leo e a da fra??o micelar, com ?ter de petr?leo, NaCl
10%(p/v) e NaSO4 2%(p/v) . Enzimas como: a-amilase, pepsina, bile, pancreatina,
lipase e mucina, bem como compostos inorg?nicos, tais como KCl, KSCN, NaH2PO4,
Na3PO4, NaOH, NaCl, CaCl2, HCl, NaHCO3. As varia??es fisiol?gicas foram
reproduzidas pelo banho de aquecimento com giro orbital (37?C) e centrifuga??o
(5000g;45 min). A quantifica??o e determina??o do perfil de caroten?ides foi realizada
por cromatografia l?quida de alta efici?ncia (CLAE), com coluna YCM? Carotenoid C30
S-3 de 4,6 x 250mm. A quantifica??o de caroten?ides totais foi realizada por
espectrofotometria UV-VIs. Todo o procedimento foi executado sob temperatura (25?C)
e luz controlada. O Beta-caroteno estava presente majoritariamente, com teores de 86%,
73% e 82%, para as variedades 1, 2 e 3 e ap?s a digest?o, o perfil do Beta-caroteno passou
a configurar teores de 96%, 89% e 100%, respectivamente. A efici?ncia de miceliza??o
foi de 23,8%, 28% e 28,9% para as variedades 1,2 e 3, indicando a transfer?ncia do -
caroteno da matriz do alimento para as micelas, correspondendo a bioacessibilidade
deste composto. Esta metodologia demonstrou-se mais r?pida e mais barata, quando
comparada aos estudos in vivo, que s?o mais onerosos, complexos e demandam mais
tempo.
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Simula??o de digest?o in vitro acoplada a modelos de transporte g?strico e intestinal para estimar a capta??o e absor??o de antocianinas em frutos / Simulation of in vitro digestion coupled to gastric and intestinal transport models to estimate the uptake and absorption of anthocyanins in fruitsPEIXOTO, Fernanda Marques 08 December 2016 (has links)
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Previous issue date: 2016-12-08 / A lot of interest in the consumption of anthocyanins increased after the association of their intake and reduced risk of chronic diseases. Despite of in vitro evidences of anthocyanins benefits to health, there is still a gap in the knowledge of the mechanisms of absorption of anthocyanins by the human body. It is known that concentration of food anthocyanins doesn't reflect the amount of these compounds which are absorbed, metabolized, distributed and biologically active in humans. Some in vitro models have been developed to evaluate the steps of cell release and transport ( uptake) of these compounds from food. The objective of this study was to evaluate the in vitro absorption of food anthocyanins using the in vitro digestion followed by uptake and transport in Caco-2 human intestinal cell line and MKN-28 human gastric cell line. Initially, anthocyanins bioaccessibility of diverse fruits was evaluated in order to select the better sources for transport assays. The bioaccessibility assays were performed using an in vitro digestion model, which mimics the human oral, gastric and intestinal stages. Quantification and characterization of anthocyanins profile were performed by high-performance liquid chromatography (HPLC) with Thermo Scientific? C1s 2.4 (4.6 x 10mm) column. After selection of the most promising fruits, the bioaccessibility tests were followed by transport assays. To assess gastric absorption, the product from gastric digestion was applied on the MKN-28 cell monolayer, which was obtained after 7 days of culture of 2.5 x 10^5 MKN-28 cells seeded in RPMI culture media in transwell? plates. The permeate was collected after 30, 60, 120 andl80 minutes oftransport. For evaluation of intestinal absorption after digestion, the digesta from the intestinal phase was applied on the Caco-2 cell monolayer, which was obtained after 21 days of culture of 2.5 x 105 Caco-2 cells seeded in DMEM culture media in TRANSWELL? plates. The permeate was collected after 30, 60 and 120 minutes of transport. All analyses were made by forming CLUE / photodiode array detector (Thermo? Scientific) at 520nm. Peel powder from jabuticaba, jambo and Jamel?o were the most promising sources. The bioaccessibility of anthocyanins after gastric digestion was 13% for jabuticaba, 45 % for jambo and 65 % for jamel?o. In addition, the intestinal bioaccessibility was 1 O % for jabuticaba, 15 % for jambo and 45 % for jamel?o. The transport assay with the MKN-28 gastric cell line, revealed 19.7%, 9.7 % and 14.1 % of transport efficiency, respectively, for jambo, jabuticaba and jamel?o digestion products. While Caco-2 intestinal cell model showed 0.8 %, 0.2 % and 0.3 % oftransport efficiency, respectively, for jambo, jabuticaba and jamel?o. These results suggest food anthocyanins are preferentially absorbed by the human gastric mucosa and to a lesser extent by the human intestinal epithelium. / O interesse pelo consumo das antocianinas aumentou ap?s o surgimento da rela??o entre o seu consumo e a redu??o do risco de doen?as cr?nicas. Apesar das evid?ncias in vitro quanto a esses beneficios ? sa?de, ainda h? uma lacuna que permanece sob investiga??o: o mecanismo de absor??o das antocianinas pelo organismo humano. Sabe-se que a quantidade desses compostos, nos alimentos, n?o reflete a quantidade absorvida, metabolizada, distribu?da e biologicamente ativa em humanos. Alguns modelos in vitro t?m sido desenvolvidos para avaliar as etapas de digest?o e transporte celular (absor??o) de compostos dos alimentos. Assim, o objetivo deste trabalho foi avaliar o transporte in vitro de antocianinas em alimentos utilizando modelos de digest?o in vitro seguido do transporte em c?lulas intestinais Caco-2 e c?lulas g?stricas MKN-28. Na 1? etapa, oito frutos foram analisados quanto aos valores de bioacessibilidade (BCSS) fornecidos pelas antocianinas presentes, para posterior sele??o para os ensaios de transporte. Os ensaios de BCSS foram realizados com um modelo de digest?o in vitro, para simula??o das fases oral, g?strica e intestinal humana. A quantifica??o e determina??o do perfil de antocianinas foram realizadas por Cromatografia l?quida de alta efici?ncia (CLAE), com coluna Thermo? Scientific C1s 2,4 (4,6 x 100mm). Na 2? etapa, realizou-se os ensaios de BCSS, anteriormente aos ensaios de transporte, nos frutos potencialmente mais promissores. Para a avalia??o do transporte g?strico, na sequ?ncia, o digerido g?strico foi aplicado sobre a monocamada de c?lulas MKN-28, com 2,5 x 10^5 c?lulas, em meio RPMI, em placa transwell? e, ap?s 7 dias de cultivo, o permeado foi coletado nos tempos 30, 60, 120, 180 minutos. Para o transporte intestinal, sequencial, o digerido intestinal foi aplicado sobre a monocamada celular Caco-2, com 2,5 x 105 c?lulas, em meio DMEM, em placas transwell? e, ap?s 21 dias de cultivo, o permeado foi coletado nos tempos 30, 60 e 120 minutos de transporte. Todas as an?lises foram realizadas por CLUE/detector de arranjo fotodiodo (Thermo? Scientific), a 520 nm. Os p?s da casca da jabuticaba, jambo e jamel?o foram as matrizes mais promissoras. A BCSS das antocianinas, ap?s a digest?o g?strica, foi de 13 % parajabuticaba, 45 % parajambo e 65 % parajamel?o, enquanto a BCSS intestinal foi de 10% para jabuticaba, 15 % para jambo e 45 % para jamel?o. Os ensaios de transporte (ET) com os modelos de c?lula MKN-28 resultaram em 19,7; 9,7 e 14,1 % de ET, respectivamente, para os p?s do jambo, jabuticaba, e jamel?o, enquanto que o modelo Caco-2, resultaram em 0,8, 0,2 e 0,3 % de ET, respectivamente. Estes resultados sugerem que as antocianinas s?o preferencialmente absorvidas pela mucosa g?strica.
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