Oral intake of cadmium, cobalt, copper, iron, lead, nickel, manganese and zinc in the university student's diet.

A duplicate diet meal study was carried out with a group of university students living in a hostel, in order to estimate the intake of Zn, Cd, Co, Cu, Fe, Mn, Ni and Pb. Zn, Cu, Fe, Mn and Ni were determined by flame atomic absorption spectrophotometry and Cd, Co and Pb by graphite furnace atomic absorption spectrophotometry after a nitric acid wet digestion procedure. The estimated intake values from the contents of breakfast, lunch, dinner and drinks were compared with the values of the Provisional Tolerable Daily Intake (PTDI) in the case of Cd and Pb, Recommended Dietary Allowances (RDA) of Co, Fe and Zn and Estimated Safe and Adequate Dietetic Daily Intake (ESADDI) of Cu and Mn. Neithe…

Isocratic high-performance liquid chromatographic determination of tryptophan in infant formulas.

The application to infant formulas of a method for tryptophan determination by isocratic HPLC with UV detection at 254 nm, after derivatization with phenyl isothiocyanate, was studied. Protein was hydrolysed by barium hydroxide at 120 degrees C for 8 h, followed by derivatization with phenyl isothiocyanate, HPLC and UV detection at 254 nm. The optimum chromatographic conditions (pH, ionic strength of elution solvent and eluent ratio) were established. The analytical parameters (linearity, precision, accuracy of derivatization and limits of detection and quantification) were determined. The values obtained demonstrated that the method is useful for determining the tryptophan content of infan…

Lactoferrin and Its Possible Role in Iron Enrichment of Infant Formulas

Se han descrito las propiedades y funciones de la lactoferrina respecto a su utilizacion en el enriquecimiento de las formulas para lactantes. Se ha prestado especial atencion a las funciones de la lactoferrina directamente relacionadas con su capacidad para fijar hierro: poder bacteriostatico, efecto sobre la absorcion de hierro y poder antioxidante. De los resultados obtenidos por diferentes autores se desprende que la adicion de lactoferrina, generalmente bovina, a las formulas para lactantes no afecta a la absorcion de hierro. No obstante, debido a su capacidad de fijacion de hierro su adicion a las formulas podria ser de utilidad por los siguientes motivos: su efecto protector en el tr…

Bioaccessibility of inorganic arsenic species in raw and cookedHizikia fusiformeseaweed

Samples of Hizikia fusiforme edible seaweed, a commercially available dried food with high concentrations of total arsenic (t-As) and inorganic arsenic (i-As), both raw and cooked (boiling at 100 °C, 20 min), were selected for the bioaccessibility study. Cooking caused a significant reduction in the concentrations of t-As (30–43%) and i-As (46–50%), despite which the i-As contents in the cooked product were high (42.7–44.6 µg g−1 seaweed). An in vitro gastrointestinal digestion (pepsin, pH 2, and pancreatin–bile extract, pH 7) was applied to the seaweed to estimate arsenic bioaccessibility (maximum soluble concentration in gastrointestinal medium) of t-As, i-As, arsenic(III) and arsenic(V).…

Biosorption of green and black tea polyphenols into Saccharomyces cerevisiae improves their bioaccessibility

Infusions of green tea (GT) and black tea (BT) and the use of Saccharomyces cerevisiae as a natural matrix were employed to check the impact of biosorption on the possible fate of tea polyphenols in the gastrointestinal tract in terms of bioaccessibility and total antioxidant capacity (TEAC and ORAC assays). The maximum biosorption yields obtained were 47.61 ± 11.57 and 99.68 ± 5.25 mg/g from GT and BT infusions, respectively. A significant increase (p < 0.05) in the recovery of phenolic compounds was shown after in vitro digestion. The bioaccessible fractions generally exhibited higher antioxidant capacities in both tea infusions and suspensions of S. cerevisiae versus non-digested samples…

High-performance liquid chromatographic determination of tocopherols in infant formulas

A method for the simultaneous determination of alpha-tocopherol acetate and alpha-, delta-, and gamma-tocopherols by normal-phase high-performance liquid chromatography (HPLC) with a fluorescent detector in infant formula is proposed. The values obtained in the determination of the analytical parameters: linearity, precision, limit of detection and accuracy (analysis of a standard reference material, SRM 1846), confirm the quality of the method. The proposed method is useful for the determination of alpha-, delta-, and gamma-tocopherols and alpha-tocopherol acetate in infant formulas at a low cost and in a total time of 2 h.

Labeling and nutritional education

Abstract Nutritional food labeling has two components, a nutrient declaration on the back-of-package (usually mandatory) and supplementary simple graphical nutrition information on the front-of-package (commonly voluntary), which are aimed at assisting consumers to be aware of the nutritional value of foods in order to make healthy food choices. This is an important tool for nutritional education that can aid consumers in the fight against diet-related chronic diseases that currently constitute a global public health concern worldwide. In this chapter, the main factors involved in the use and understanding of nutrition labels by consumers, and a detailed explanation of the characteristics o…

Effects of different infant formula components on calcium dialysability

The in vitro methods proposed for estimating calcium bioavailability can be useful in studying the effect of dietetic factors. The aim of our work was to study the possible influence of different infant formula components – calcium, phosphorus, lactose, citric acid, ascorbic acid and proteins – on calcium bioavailability, considering dialysability to be an estimate of bioavailability. The 1981 method of Miller et al. with slight modifications (concerted action no 10 – FLAIR project) was applied to 18 commercial infant formulas of six different types: adapted, follow-up, preterm, hydrolysates, lactose-free and soy-based. Significant positive linear correlations between the amount of calcium …

Apoptotic effect of a phytosterol-ingredient and its main phytosterol (β-sitosterol) in human cancer cell lines

Dietary interventions may effectively control cancer development, with phytosterols (PS) being a class of cancer chemopreventive dietary phytochemicals. The present study, for the first time, evaluates the antiproliferative effects of a PS-ingredient used for the enrichment of several foods and its main PS, β-sitosterol, at physiological serum levels, in the most prevalent cancer cells in women (breast (MCF-7), colon (HCT116) and cervical (HeLa)). In all three cell lines, these compounds induced significant cell viability reduction without a clear time- and dose-dependent response. Moreover, all treatments produced apoptotic cell death with the induction of DNA fragmentation through the app…

Antioxidant effect derived from bioaccessible fractions of fruit beverages against H2O2-induced oxidative stress in Caco-2 cells

Abstract This work evaluates the effect of bioaccessible fractions from fruit beverages against oxidative stress (OS) in Caco-2 cells. A fruit beverage (grape + orange + apricot) (with/without milk and/or iron/zinc) was subjected to in vitro gastrointestinal digestion, and bioaccessible fractions were incubated with Caco-2 cell cultures. Following preincubation, OS was induced with 5 mM H 2 O 2 . Intracellular reactive oxygen species (ROS), mitochondrial potential (Δ ψ m ), mitochondrial metabolism (MTT test), intracellular reduced glutathione (GSH) and superoxide dismutase activity (SOD) were measured. The data evidenced viable cultures with increased mitochondrial metabolism and GSH-Rd ac…

Impact of high-pressure processing on the stability and bioaccessibility of bioactive compounds in Clementine mandarin juice and its cytoprotective effect on Caco-2 cells.

Mandarin juice is a rich source of antioxidant bioactive compounds. While the content and profile of bioactives are known, the impact of high-pressure processing (HPP) on their stability and bioaccessibility (BA) is unknown, but may allow obtaining safe, nutritious, and fresh-tasting juices with highly extractable bioactive compounds. The stability and BA of bioactive antioxidant compounds in untreated and HPP-treated (400 MPa/40 °C/1 min) Clementine mandarin juices, and the cytoprotective effect of its bioaccessible fractions (BF) obtained after simulated gastrointestinal digestion against H2O2-induced oxidative stress in differentiated Caco-2 cells were investigated. The BF of HPP-treated…

The impact of galactooligosaccharides on the bioaccessibility of sterols in a plant sterol-enriched beverage: adaptation of the harmonized INFOGEST digestion method.

The effect of the addition of galactooligosaccharides (GOS) on sterol bioaccessibility in three plant sterol (PS)-enriched milk-based fruit beverages (without GOS addition (MfB) and with 2.5 g (MfB-G2) and 5.0 g (MfB-G5) GOS per 250 mL) was evaluated after micellar gastrointestinal digestion. Cholesterol bioaccessibility was very similar among beverages, though a slight significant increase (from 80% to 85%) was observed by the addition of 5.0 g GOS. The addition of GOS did not affect total PS bioaccessibility (≈37%). Based on the results obtained after micellar digestion, it has been demonstrated that these beverages could be a suitable food matrix for simultaneous enrichment with PS and G…

Availability of iron from milk-based formulas and fruit juices containing milk and cereals estimated by in vitro methods (solubility, dialysability) and uptake and transport by Caco-2 cells

Abstract Iron solubility, dialysability and transport and uptake (retention + transport) by Caco-2 cells as indicators of iron availability have been estimated in the in vitro gastrointestinal digests of infant foods (adapted, follow-up and toddler milk-based formulas and fruit juices containing milk and cereals (FMC)). Low correlation coefficients (in all cases R -squared ⩽ 37.1%) were obtained between iron solubility or dialysability versus transport or uptake efficiency – a fact emphasizing the importance of incorporating Caco-2 cell cultures to in vitro systems in order to adapt the conditions to those found in in vivo assays. The highest uptake efficiency corresponded to FMC (25.6–26.1…

Influence of storage and in vitro gastrointestinal digestion on totqal antioxidant capacity of fruit beverages

Eight fruit beverages containing grape, orange and apricot, with/without iron and/or zinc and with/without milk added were analyzed for total antioxidant capacity (ORAC and TEAC methods), ascorbic acid content, and total polyphenols. The influence of cold storage (2-4 degrees C) during the product shelf-life (135 days) and antioxidant capacity after an in vitro gastrointestinal digestion were evaluated. Antioxidant capacity for all beverages together increased significantly (p < 0.05) at the end of storage (16.4% and 12.8% for ORAC and TEAC, respectively), whereas ascorbic acid remained stable. Regarding in vitro digestion, antioxidant values of bioaccessible fractions of fruit beverages in…

Evaluation of the Cytotoxicity of Cholesterol Oxides in Human Colon Cancer Caco-2 Cells

The content of cholesterol oxides (COPs) in foods varies between 0.1 and 294.3 mg/g. These oxides are formed by auto-oxidative enzymatic processes promoted by the heat treatment of food and/or to exposition of them to the presence of oxygen and sunlight during storage. Their importance is that they are associated with pathological processes like apoptosis, dyslipidemia and pro-oxidative states, among others. The objective was to evaluate the cytotoxicity, by means MTT assay, of 7-keto cholesterol (7KC), cholestane-triol (Triol), α-epoxy cholesterol (α-epoxy C), β-epoxy cholesterol (β-epoxy C) in differentiated Caco-2 cells at 120μM for 24, 48 and 72h.Triol was the most cytotoxic COP, with a…

Bioaccessibility of calcium, iron and zinc from three legume samples

Legumes can be a source of mineral elements but also of antinutritional factors which can affect their absorption. An in vitro method including gastrointestinal digestion was used to estimate mineral bioavailability. Soluble (bioaccessible) and insoluble calcium, iron and zinc from white beans, chickpeas and lentils were determined after gastrointestinal digestion. The influence of the original sample weight on the soluble mineral fraction was also estimated. The results obtained show that white beans are the legumes with the highest bioaccessible calcium and iron contents. Lentils have a high iron content but its bioaccessibility is much lower than that of iron from white beans and chickpe…

Determination of mercury in dry-fish samples by microwave digestion and flow injection analysis system cold vapor atomic absorption spectrometry

Abstract Flow injection analysis system cold vapor atomic absorption spectrometry (FIAS-CV-AAS) preceded by a wet digestion in a microwave oven, as a method for measuring mercury in fish was studied. The digestion process and conditions of the FIAS (carrier concentration: HCl 3% v v ; reducing agent: SnCl2 2% w v : filling and injection times: 8 and 25 min, respectively; and sample volumes) were optimized. The analytical parameters of the proposed method (detection limit = 7.7 ng/g; precision intraassay = 6.7%; interassay = 14.0%) demonstrates its adequacy and are similar to the ones (detection limit = 19.4 ng/g; precision intraassay = 11.2%; interassay = 15.9%) obtained using a conventiona…

7keto-stigmasterol and 7keto-cholesterol induce differential proteome changes to intestinal epitelial (Caco-2) cells

Abstract Recent studies have expanded the appreciation of the roles of oxysterols triggering inflammatory, immune cytotoxic and apoptotic processes, but have not been considered for proteome analysis. A comparative proteomic study in intestinal epithelial cell cultures incubated (60 μM/24 h) with 7keto-cholesterol or 7keto-stigmasterol was performed. The influence of both compounds was studied following the nLC-TripleTOF analysis. Findings were compared to results for control cultures. In the principal component analysis (PCA) of proteome patterns, two components were extracted accounting for 99.8% of the variance in the protein expression. PCA analysis clearly discriminated between the per…

Stability of fatty acids and tocopherols during cold storage of human milk

The stability of fatty acids and tocopherols from human milk from 8 healthy lactating mothers was determined in fresh milk and after cold storage. Refrigeration at 4 °C for 48 h or freezing at −20 °C for 30 days did not significantly decrease fatty acid contents (mg per 100 g human milk), or concentrations of α-, β-, γ- and δ-tocopherol isomers, compared with fresh milk. Results obtained in the present study showed that cold storage under the usual storage conditions in neonatal units, at home or in milk banks did not have a detrimental effect upon fatty acids and tocopherols contents in human milk. Thus, infants can receive stored human milk without losses in the nutritional value of these…

GFAAS determination of selenium in infant formulas using a microwave digestion method.

A method for determining the selenium content of infant formulas is proposed. It includes wet digestion with nitric acid and hydrogen peroxide in medium pressure teflon bombs in a microwave oven and determination by graphite furnace atomic absorption spectrometry (GFAAS). The absence of interferences is checked. Values obtained for the limit of detection (19.4 ng/g), precision (RSD = 2.2%) and accuracy by analysis of a reference material show that the method is reliable.

Uptake and retention of calcium, iron, and zinc from raw legumes and the effect of cooking on lentils in Caco-2 cells

Abstract This study examined calcium, iron, and zinc uptake in Caco-2 cells (retention plus transport) from white beans, chickpeas, and lentils subjected to prior in vitro gastrointestinal digestion and the effect of cooking (traditional and industrial—ready to eat) on the uptake of these minerals from lentils. The highest cell uptake of calcium, iron, and zinc corresponded to raw chickpeas, which had the lowest soluble oxalate content and intermediate phenolic and tannin contents. From these results, raw chickpeas would be the best dietary source of calcium, iron, and zinc, although consumption in this form (ie, raw) is low. Cooking affects the calcium, iron, and zinc contents of lentils a…

Determination of sialic acid and gangliosides in biological samples and dairy products: A review

Gangliosides are sphingolipids containing one or more moieties of sialic acid in their structure. Both gangliosides and sialic acid are bioactive compounds related to animal physiology. Due to their biological relevance, analytical methods adapted to each type of matrix have been developed over time. The present study reviews the main methods applied to the analysis of sialic acid and gangliosides in biological samples and dairy products.

Dialyzability of iron, zinc, and copper of different types of infant formulas marketed in Spain

The bioavailability of trace elements in infant formulas is affected by different physiological and dietetic factors. In vitro methods based on element dialyzability have been proposed to estimate the bioavailability. Infant formulas of the same type but from different manufacturers can differ in the salt used for supplementation and in the contents of other components that can affect mineral bioavailability. The aim of our study is to estimate the dialyzability of iron, zinc, and copper of formulas marketed in Spain, in order to detect possible differences in formulas of the same type coming from different manufacturers. At the same time, the effects of the type of formula, the composition…

Effects of Plant Sterols or β-Cryptoxanthin at Physiological Serum Concentrations on Suicidal Erythrocyte Death.

The eryptotic and hemolytic effects of a phytosterol (PS) mixture (β-sitosterol, campesterol, stigmasterol) or β-cryptoxanthin (β-Cx) at physiological serum concentration and their effect against oxidative stress induced by tert-butylhydroperoxide (tBOOH) (75 and 300 μM) were evaluated. β-Cryptoxanthin produced an increase in eryptotic cells, cell volume, hemolysis, and glutathione depletion (GSH) without ROS overproduction and intracellular Ca2+influx. Co-incubation of both bioactive compounds protected against β-Cx-induced eryptosis. Under tBOOH stress, PS prevented eryptosis, reducing Ca2+influx, ROS overproduction and GSH depletion at 75 μM, and hemolysis at both tBOOH concentrations. β…

Effects of legume processing on calcium, iron and zinc contents and dialysabilities

Legumes are a good source of calcium, iron and zinc, but are also a source of phytates and dietary fibre components that can negatively affect the bioavailability of these minerals. To estimate the latter, an in vitro dialysis method can be applied that gives the dialysability of a mineral as an estimate of its availability for absorption. Calcium, iron and zinc contents and dialysabilities in three legumes (beans, chickpeas and lentils) and the effects of cooking treatments and industrial processing on these parameters were studied. Beans had the highest calcium content (1.54 g kg -1 dry matter (DM)) and chickpeas the lowest iron content (46.9mg kg -1 DM), whilst the zinc contents were sim…

Determinación del colesterol en leche humana: una alternativa a los métodos cromatográficos

Introduction: human milk (HM) is considered the best option for feeding healthy infants. Cholesterol (CHOL) is important for proper development of the nervous system, and for hormone and vitamin synthesis in growing infants. Breastfeeding and dietary CHOL intake during infancy have been suggested to affect blood lipid levels and the risk of cardiovascular disease in adulthood. Gas chromatography is the technique most widely used to determine CHOL in HM. Chromatographic methods are specific for the determination of CHOL and other sterols present in HM, but are extremely time consuming, and the costs and equipment requirements mean that they are not accessible to all laboratories. Aim: the pr…

Sterol stability in functional fruit beverages enriched with different plant sterol sources

Abstract Two different plant sterol (PS) sources (free PS from tall oil and esterified PS from vegetable oils) were used for manufacturing two types of functional beverages (fruit and milk-based fruit beverages), and their PS and phytosterol oxidation product (POP) contents were determined. Gas chromatography–tandem mass spectrometry (GC–MS/MS) was used for identification and gas chromatography–flame ionization detection (GC–FID) for quantitation purposes. Brassicasterol, campesterol, campestanol, stigmasterol, β-sitosterol and sitostanol were the quantified PS, conforming a profile in order with current legislation. The relative percentages of PS differed according to the enrichment source…

In vitrobioaccessibility of iron and zinc in fortified fruit beverages

Summary Iron and zinc bioaccessibility was estimated in the in vitro gastrointestinal digests of six different fortified fruit beverages (Fb) containing iron and/or zinc and/or skimmed milk (M). Solubility values can be used to establish trends in relative bioavailability of iron and zinc, as the first stage towards mineral bioavailability comprises solubility in the intestinal tract. FbFe, FbFeM and FbFeZnM samples showed iron bioaccessibility above 88%, differing (P < 0.05) from those of FbFeZn (53%). In turn, FbZn, FbFeZn and FbZnM samples presented higher zinc bioaccessibility (above 68%), differing (P < 0.05) from those of FbFeZnM (48%). The presence of milk-derived caseinophosphopepti…

Stability of tocopherols in adapted milk-based infant formulas during storage

Abstract An evaluation is made of the effects of added α -tocopherol ( α -T) or α -tocopherol acetate ( α -TAc) combined with iron lactate or sulfate, storage time (up to 17 months) and storage temperature (22°C and 37°C) on the α -TAc, α -T, γ -tocopherol ( γ -T) and δ -tocopherol ( δ -T) content of infant formulas. Tocopherols were measured by normal phase HPLC and fluorescence detection after direct extraction, which allows the determination of both α -TAc and α -T. The α -TAc content was not affected by temperature or storage time, although the α -, γ - and δ -T contents, which were lower at 37°C than at 22°C, were significantly affected. The type of iron salt added (lactate or sulfate)…

Impact of Lipid Components and Emulsifiers on Plant Sterols Bioaccessibility from Milk-Based Fruit Beverages.

Sterol bioaccessibility (BA) of three plant sterol (PS)-enriched milk-based fruit beverages (MFb) with different fat contents (1.1–2.4%), lipid sources (animal or vegetable), and without or with emulsifiers (whey proteins enriched with milk fat globule membrane (MFGM) or soy lecithin) was evaluated after simulated gastrointestinal digestion. The BA of total PS followed the order 31.4% (MFbM containing milk fat and whey proteins enriched with MFGM) = 28.2% (MFbO containing extra virgin olive oil and soy lecithin) > 8.7% (MFb without fat addition). Total and individual PS content in the bioaccessible fractions followed the order MFbM > MFbO > MFb. Consequently, formulation with MFGM is propos…

Effect of lactoferrin addition on the dialysability of iron from infant formulas

A possible enhancing effect of lactoferrin (Lf) on iron absorption by breast-fed infants has been suggested, however the available results failed to confirm this hypothesis. Nevertheless, Lf could be useful in protecting the lipid fraction of infant formulas against oxidation. Concerning the possibility of adding Lf to infant formulas with this aim, we considered it necessary to evaluate the effect of this addition on iron dialysability, which was used as a parameter indicator of bioavailability. An in vitro dialysability method was applied to three types of infant formulas, with and without Lf added, respectively. In none of the analysed formulas did the added Lf have a negative effect on …

Calcium, iron and zinc uptakes by Caco-2 cells from white beans and effect of cooking

White beans (Phaseolus vulgaris L.) have an interesting content of essential elements, calcium, iron and zinc, but they content also phytates, oxalates, proteins, polyyphenols and complex polysaccharides that are known to interact with minerals and to affect their bioavailability. The bioavailability of calcium, iron and zinc from raw and cooked white beans was estimated using their uptake by Caco-2 cells as the criteria. Previously, the mineral fraction (soluble or dialysable) to be added to the Caco-2 cell monolayer was selected. The results obtained show that cooking increases the Caco-2 cells' uptake percentages (calcium, 18.8 versus 3.6; iron, 33.7 versus 1.7; and zinc, 17.2 versus 2.1…

Does the addition of caseinophosphopeptides or milk improve zinc in vitro bioavailability in fruit beverages?

Abstract The influence of caseinophosphopeptides (CPPs) added to fruit beverage versus milk based fruit beverage upon zinc retention, transport, and uptake, as well as the influence of Fe supplementation, were studied using a combined simulated gastrointestinal digestion/Caco-2 cell system. Zinc retention, transport, and uptake of milk based fruit beverage was 4- to 5-fold greater than that of fruit beverages with or without CPPs – no statistically significant differences being observed in relation to the presence or absence of CPPs. Possibly, a slow release of CPPs throughout the digestive tract, as can be expected to take place during the digestion of casein, has a more beneficial effect …

Determination of Plasma Lipid Hydroperoxides by an NADPH/NADP + Coupled Enzyme Reaction System. Evaluation of a Method

Summary: Several techniques based on different principles have been proposed to measure lipid hydroperoxides. Enzymatic methods are sensitive and can be quite specific but they are subject to interference by inhibitors and not all are stoichiometric. The present work proposes some modifications of the Heath & Tappel (Anal Biochem 1976; 7:184—91) enzymatic method of determination of lipid hydroperoxides in order to standardize and automate it and to meet the analytical criteria required for a biological assay. The proposed new protocol and the automated assay give acceptable within-run and between-run precisions, with coefficients of variation of 3.34% and 5.80%, respectively, at the usual p…

Identification of Casein Phosphopeptides in β-casein and Commercial Hydrolysed Casein by Mass Spectrometry

Casein phosphopeptides (CPPs) in commercial hydrolysed casein (CE90CPP) and in β-CN (β-CN) after simulated gastrointestinal digestion (gastric stage pepsin, pH =2, 37°C 2h) and intestinal stage (pancreatic-bile extract, pH =5.2, 37°C 2h) were sequenced by on-line reversed-phase high performance liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (RP-HPLC-ESIMS/MS). In β-CN digest five peptides that contained four to five phosphate groups and the cluster sequence SpSpSpEE (residues 17-21) were identified. All CPPs with one exception β-CN(1-24)4P, had the protein fragment β-CN(1-25)4P, which is one of the main CPPs produced in vivo digestion of casein and the re…

Antiproliferative activity of green, black tea and olive leaves polyphenols subjected to biosorption and in vitro gastrointestinal digestion in Caco-2 cells.

Olive (Olea europaea L.) leaves and tea (Camellia sinensis) are rich sources of bioactive compounds, especially polyphenols. Our previous studies have evidenced the potential use of Saccharomyces cerevisiae as a natural delivery system for these antioxidants and a means to improve their bioaccessibility in the human gut. In the present work, the antiproliferative effect of green tea (GT), black tea (BT) and olive leaves (OL) infusions and suspensions of S. cerevisiae were evaluated, for the first time, in human colon cancer cells (Caco-2) after biosorption and in vitro gastrointestinal digestion. The bioaccessible fractions (BF) were not overtly cytotoxic, not affecting cell viability. ROS …

Comparison of spectrophotometric and HPLC methods for determining sialic acid in infant formulas

Abstract Two methods for determining sialic acid in infant formulas – spectrophotometry and HPLC with fluorescence detection – have been optimised and validated, the first one allows to determine total sialic acid while the second allows to differentiate the two main forms of sialic acid (N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc)). A common sample preparation procedure (hydrolysis and purification) for both methods has been proposed. The linearity (from 6 to 150 μg of total sialic acid in the assay for spectrophotometry, and from 12.5 to 250 ng and 1 to 5 ng of Neu5Ac and Neu5Gc, respectively, for HPLC) is adequate. The detection and quantification limits (0.29…

Protective effect of antioxidants contained in milk-based fruit beverages against sterol oxidation products

Abstract Sterol oxidation products (SOPs) have shown cytotoxic effect in human intestinal cells; however, their effect within a food matrix has not been assayed yet. This study evaluated the possible cytotoxic effect of SOPs within bioaccessible fractions (BFs) of two milk-based fruit beverages with (BFA)/without (BFB) plant sterols in differentiated Caco-2 cells and if the BFs counteracted the cytotoxic effect induced by COPs mixture (30 and 60 μM). BFs did not evoke cytotoxic effect in any of the tests carried out and they protected against the loss of intestinal cohesion, mitochondrial depolarization and necrosis induced by COPs mixture. Moreover, BFB sample protected from cell cycle arr…

Cytokines profiles in intestinal epithelial (Caco-2) cells exposed to 7-ketostigmasterol or 7-ketocholesterol

Identification of Novel Phosphopeptides After Simulated Digestion of αs2-casein by Tandem Mass Spectrometry

Casein phosphopeptides (CPPs) are encrypted in αs1-, αs2-and β-casein (CN) and can be released by in vitro, in vivohydrolysis or food processing of dairy foods. Bovine αs2-CN contains two cluster sequences of anionic phosphoseryl and glutamyl residues SpSpSpEE in its structure (residues 8–12 and 56–63), which can modulate mineral bioavailability. In this study αs2-casein (αs2-CN) was subjected to simulated gastrointestinal digestion. CPPs released were sequenced by on-line reversed-phase high performance liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (RP-HPLC-ESIMS/MS). Six novel αs2-CN derived CPPs, Three of them (αs2-CN(54–87)4P,αs2-CN(24–70)4P and αs2-…

Iron bioavailability in iron-fortified cereal foods: The contribution of in vitro studies

Iron deficiency anemia is the most common nutritional deficiency in humans. Not all dietary ingested iron, heme or nonheme, will be available to absorption and negative imbalance between iron requirements and absorption leads to iron deficiency and/or anemia. The recommended iron values usually are based on the genetic and on diet iron-bioavailability, which can be considered as the principal factor that change among the cultures and influences the distinct levels of recommendation among countries. Dietary changes present practical limitations due to be difficult to change food habits. The iron food fortification is considered more cost effective and economically more attractive than iron s…

A positive impact on the serum lipid profile and cytokines after the consumption of a plant sterol-enriched beverage with a milk fat globule membrane: a clinical study

The hypocholesterolemic effect and the modification of serum biomarkers of a dietary plant sterol (PS) intake, cholesterol precursors and cytokines after the consumption of milk-based fruit beverages with a milk fat globule membrane were evaluated by a randomized, double-blind, crossover, multiple dose bioavailability study. Postmenopausal women (n = 38) consumed daily 250 mL of a beverage with or without 2 g of PS added during 6 weeks in each of the study periods. With the intake of the PS-added beverage, significant decreases (mg dL-1) in serum total cholesterol (pre-treatment: 220.0 ± 27.8 vs. post-treatment: 212.9 ± 25.8; p < 0.05) and LDL-cholesterol (129.4 ± 28.5 vs. 121.7 ± 24.4; p <…

Whole blood selenium content in pregnant women

The selenium content in the blood of pregnant women in the Valencian Community, Spain was determined until the end of gestation in order to assess its evolution and detect possible differences in relation to the values corresponding to non-pregnant women of fertile age. A total of 158 blood samples were obtained from 137 pregnant women. Samples were classified as: (a) first; (b) second; and (c) third trimester. Selenium was determined by a flow injection hydride atomic absorption spectrometric method (Seronorm whole blood was used as a quality control check). The selenium values obtained for the three trimesters followed a Gaussian distribution. The intervals for mean found were: (a) 75.7-8…

Effect of simulated gastrointestinal digestion on sialic acid and gangliosides present in human milk and infant formulas.

The effects of simulated gastrointestinal digestion upon sialic acid and gangliosides in infant and follow-on formulas and human milk, as well as their bioaccessibility, have been evaluated. The gastric stage is the step that causes a greater decrease in sialic acid and ganglioside contents. The intestinal stage only decreases the total and individual contents of gangliosides. After gastrointestinal digestion, neither sialic acid nor gangliosides were found in the nonbioaccessible fraction. The highest bioaccessibility (100 × content in soluble fraction after gastrointestinal digestion/total content) of sialic acid is found in human milk (87%), followed by infant formula (77%) and follow-on…

Direct determination of lead in cola beverages by electrothermal atomic absorption spectrophotometry

Amino Acid Contents of Infant Formulas

Abstract The amino acid supply in an infant's first month of life must be sufficient in quantity and quality to fulfil the needs of this period of life. Guidelines, recommendations and minimum and maximum values have been established regarding to amino acid composition of infant formulas. The aim of this study was to determine the amino acid profile of infant formulas of different types marketed in Spain in order to assess the compliance with the recommendations (ESPGAN) and to detect possible differences in the amino acid profiles depending on the protein source. Amino acids were determined by HPLC in ion-pair reversed phase after hydrolysis and derivatization with phenylisothiocyanate. Ne…

Bioavailability of zinc from infant foods byin vitro methods (solubility, dialyzability and uptake and transport by Caco-2 cells)

The aim of this study was to evaluate the bioavailability of zinc from infant foods (adapted, follow-up and toddler milk-based formulas and fruit juices containing milk and cereals, FMC) using solubility, dialyzability and a model combining simulated gastrointestinal digestion and zinc uptake and transport by Caco-2 cells. The greater solubility of zinc from infant formulas compared with fruit juices (FMC) could be due to the greater casein phosphopeptide content resulting from casein hydrolysis. The highest zinc dialysis percentage corresponded to FMC, which on the other hand had the lowest zinc contents of the analyzed samples. The presence of organic acids in samples of this kind favors …

COPPER | Properties and Determination

Direct determination of calcium, magnesium, sodium, potassium and iron in infant formulas by atomic spectroscopy. Comparison with dry and wet digestions methods

An optimization and comparison of three atomic absorption spectroscopic methods for measuring Ca, Mg, Na, K and Fe contents of infant formulas are carried out. The first method is a direct measure of a sample dispersion in water. The second method involves a previous wet digestion (HNO3 + H2O2) in a microwave oven. And finally a dry mineralization at 450 °C is assayed. In order to measure Ca and Mg lanthanum at 0.4% concentration and in the case of Na and K cesium at 0.1% were added. The analytical parameters of the three methods assayed were determined. Results obtained show that the three studied methods are accurace and reliable and no significant difference among them at a probability l…

Improved bioaccessibility and antioxidant capacity of olive leaf ( Olea europaea L.) polyphenols through biosorption on Saccharomyces cerevisiae

Abstract Olive ( Olea europaea L.) leaves (OL) are natural by-products that can be used as an advantageous rich source of bioactive compounds, especially polyphenols. This study investigated the potential use of Saccharomyces cerevisiae as a new carrier of OL antioxidants and a means to protect their bioactivity during simulated gastrointestinal digestion. The biosorption on S. cerevisiae allowed the recovery of 25.17 mg/g and 49.40% of the polyphenols from olive leaf (var. Chemlali ) infusions. Both infusions and suspensions before and after biosorption were analyzed for total soluble polyphenols and total antioxidant capacity (oxygen radical absorbance capacity (ORAC) and trolox equivalen…

In vitro interactions between calcium, zinc, copper and iron in milk- and soy-based infant formulas / Interacciones in vitro entre calcio, cinc, cobre e hierro en formulas de base láctea y de soja para lactantes

Interactions among calcium, zinc, copper and iron in milk- and soy-based infant formulas were as sessed in order to evaluate the influence of supplementation with one trace element on the bioavailability of the others. The criterion used to estimate the bioavailability is the percentage of dialysis, estimated by applying an in vitro method. Infant formulas were supplemented with CaCl2, ZnO, CuCO3 Cu(OH) 22H2O and FeSO4 7H2O at two levels (maximum (+) and minimum (-) allowed by the European Union). A factorial design at two levels with three factors (Zn, Cu and Fe addition) was carried out. The main effects and the second-order interactions of the studied factors on the bioavailability of a…

Impact of Fruit Beverage Consumption on the Antioxidant Status in Healthy Women

&lt;i&gt;Background:&lt;/i&gt; Epidemiologic studies suggest that antioxidant-rich foods might reduce the risk of cancer and cardiovascular diseases. &lt;i&gt;Aim:&lt;/i&gt; To test the health-protective potential of three fruit beverages, Fb (grape-orange-apricot), FbM (Fb with skimmed milk) and FbMFe [FbM + Fe(II)], in healthy women. &lt;i&gt;Methods:&lt;/i&gt; The influence of fruit beverage consumption (500 ml/day) upon serum antioxidant capacity determined by ORAC and TEAC methods and erythrocyte superoxide dismutase (SOD) activity was assessed in 32 healthy female volunteers. In the intervention study, each subject received the fruit beverages during three periods (3 weeks for Fb and …

Selenium content of infant formulas and estimation of the intake of bottle fed infants.

Calcium, magnesium, sodium, potassium and iron content of infant formulas and estimated daily intakes

The calcium, magnesium, sodium, potassium and iron content of a total of 22 different infant formulas marketed in Spain were measured by atomic spectrometry, and the mineral intake of infants fed exclusively with these formulas was estimated. The contents (mg/100kJ) are in the following ranges: Ca, 14-24; Mg, 1.1-2.8; Na, 5.6-9.8; K, 19-35; Fe, 0.02-0.50. These values coincide with those recommended by the Codex and European Society for Paediatric Gastroenterology and Nutrition (ESPGAN), and do not exceed the limits established by the European Union (EU). The mean values and ranges of estimated intakes for each formula type and period of infancy (0-1, 1-2, 2-3, 3-4 and 4-5 months) expressed…

Effect of plant sterol and galactooligosaccharides enriched beverages on oxidative stress and longevity in Caenorhabditis elegans

Abstract This study evaluates the impact of two plant sterol (PS) enriched fruit beverages (0.6 g /100 ml), without (MfB) or with GOS (MfB-G) (1.2 g/100 ml) on the resistance against oxidative stress induced by non-conventional (cholesterol oxidation products (COPs)) and conventional (H2O2) oxidant compounds, as well as their impact on C. elegans longevity. Nematodes fed with both beverages (0.005–0.25%, v/v) showed similar improved oxidative stress resistance against COPs and H2O2. This effect was dependent on daf-16 transcription factor, although GOS showed an additional beneficial effect independent to daf-16. In addition, both beverages extended nematode lifespan, independently of the p…

Environmental cadmium, lead and nickel contamination: possible relationship between soil and vegetable content

The cadmium, lead and nickel content of soils of four agricultural areas exposed to different degrees of environmental pollution and vegetables grown there were measured by Atomic Absorption Spectrophotometry (AAS). In order to ascertain the possible relationship between the heavy metal content of soils (total and extractable) and of vegetables grown in them, the correlation between the two was calculated. The highest correlation values between metal content of soil and vegetables are these corresponding to nickel: the total metal content in soils and leaves-stems are linear (p<0.01). The extractable metal content of soils and leaves-stems are linear (p<0.1). Therefore, soil seems to be the…

Evaluation of the Bioaccessibility of Antioxidant Bioactive Compounds and Minerals of Four Genotypes of Brassicaceae Microgreens

Microgreens constitute an emerging class of fresh, healthy foods due to their nutritional composition. In this study the content of minerals and antioxidant bioactive compounds, and for the first time bioaccessibility, were evaluated in broccoli (Brassica oleracea L. var. italica Plenck), green curly kale (Brassica oleracea var. sabellica L.), red mustard (Brassica juncea (L.) Czern.) and radish (Raphanus sativus L.) hydroponic microgreens. Macro- (K, Ca, Mg) and oligo-elements (Fe, Zn), ascorbic acid, total soluble polyphenols, total carotenoids, total anthocyanins, total isothiocyanates and total antioxidant capacity (Trolox Equivalent Antioxidant Capacity and Oxygen Radical Absorbance Ca…

CHEMICAL ANALYSIS FOR SPECIFIC COMPONENTS | Micronutrients and Other Minor Meat Components

The chemical analysis of the many micronutrients (minerals and vitamins) and other components such as cholesterol and enzymes in meat and meat products are addressed in this article. The sample preparation and main methods for the determination of these components are reviewed and described as well as their respective major advantages and disadvantages.

Determination of cholesterol in human milk: an alternative to chromatographic methods

Introduction: human milk (HM) is considered the best option for feeding healthy infants. Cholesterol (CHOL) is important for proper development of the nervous system, and for hormone and vitamin synthesis in growing infants. Breastfeeding and dietary CHOL intake during infancy have been suggested to affect blood lipid levels and the risk of cardiovascular disease in adulthood. Gas chromatography is the technique most widely used to determine CHOL in HM. Chromatographic methods are specific for the determination of CHOL and other sterols present in HM, but are extremely time consuming, and the costs and equipment requirements mean that they are not accessible to all laboratories. Aim: the pr…

Antiproliferative effect of plant sterols at colonic concentrations on Caco-2 cells

Abstract Plant sterols (PS) have been incorporated to foods due to their cholesterol-lowering effect. Because of their low intestinal absorption (0.5–2%), they can reach the colon and exert local actions. The aim of this study was to evaluate the antiproliferative effect of individual (β-sitosterol, campesterol and stigmasterol) and combined PS in colon cancer cells (Caco-2) at human colonic concentrations after simulated gastrointestinal digestion of a PS enriched milk-based fruit beverage. β-Sitosterol, campesterol and stigmasterol induced significant cell viability reduction (13–59% vs control), but only stigmasterol produced an overproduction of reactive oxygen species (92% vs control).…

Methylmercury and inorganic mercury determination in fish by cold vapour generation atomic absorption spectrometry

Abstract Given that organic mercury is more dangerous than the inorganic form and that it is converted into methylmercury by biological methylation, we have studied and optimized a simple method for measuring both organic and inorganic mercury contents in fish, using a spectroscopic vapour generation technique, with a sequential reduction of the digested sample with stannous chloride and sodium tetrahydroborate. Prior to applying the method the sample was subjected to alkaline wet digestion. Due to the matrix interferences calibration curves with matrix addition were needed for mercury determinations. The analytical parameters of the method were: linearity from 10 to 200 ng of Hg in the red…

Direct determination of lead in human milk by electrothermal atomic absorption spectrometry

Abstract Infants are particularly sensitive to the toxic effects of lead. Since milk is their only or main food it is important to know the contribution of milk to lead intake. The purpose of this study was to develop a direct method for determining the lead content of human milk by electrothermal atomic absorption spectrometry when a deuterium lamp is the only background correction available. The optimum conditions for lead determination in breast milk: sample dilution in Triton ×-100, modifier (75 μg of palladium) and nitric acid contents (2%) and the graphite furnace program (mineralization: 1100°C; atomization: 2300°C) were selected. The analytical parameters are: linearity (20–300 ng m…

Cytotoxic effect of As(III) in Caco-2 cells and evaluation of its human intestinal permeability.

Inorganic arsenic has been classified as a carcinogen for humans (Group I). However, its transit across the human intestinal epithelium has not been characterized. Using Caco-2 cells, the thiol-redox balance and apparent permeability coefficients (P(app)) for As(III) in the apical to basolateral (AP-BL) and basolateral to apical (BL-AP) direction were evaluated. After As(III) exposure, GSH-induced synthesis was observed, increasing the GSH/GSSG ratio by elevating the As(III) concentration. The AP-BL permeabilities decreased as the As(III) concentrations increased, indicating the existence of a mediated transport mechanism. The (BL-AP)/(AP-BL) permeability ratios were higher than unity, sugg…

Milk versus caseinophosphopeptides added to fruit beverage: Resistance and release from simulated gastrointestinal digestion

The influence of simulated gastrointestinal digestion on caseinophosphopeptides (CPPs) formation in milk-based fruit beverage was evaluated, together with resistance of a pool of CPPs added to fruit beverage. In milk-based fruit beverage, four CPPs were identified that can be justified by their presence in raw milk or due to processing. When it was subjected to simulated gastrointestinal digestion, 10 CPPs were identified, and only 1 presented the cluster (SpSpSpEE) (3 phosphoseryl group followed by 2 glutamic acid residues), which corresponded to αs2-CN(1-19)4P. CPPs added to fruit beverage are resistant to simulated gastrointestinal digestion, and 16 CPPs were identified originating from …

Evaluation of methylmercury cytotoxicity at intestinal level

Development of Functional Beverages: The Case of Plant Sterol-Enriched Milk-Based Fruit Beverages

Abstract Beverages are by far the most active functional food category, since they are excellent means for delivering nutrients and bioactive compounds such as plant sterols (PS). These phytochemicals are used as functional food ingredients due to their well-known cholesterol-lowering activity. The PS-enriched milk-based fruit beverages seem appropriate for complying with health recommendations (restrictions of saturated fatty acids, cholesterol, and energy value), as good sources of other bioactive compounds (polyphenols, carotenoids, and vitamins), and as a good option for obtaining the recommended daily amount of PS in subjects with moderate hypercholesterolemia. This chapter focuses on …

Antiproliferative Effect of Bioaccessible Fractions of Four Brassicaceae Microgreens on Human Colon Cancer Cells Linked to Their Phytochemical Composition

The antiproliferative effect of the bioaccessible fractions (BFs) of four hydroponic Brassicaceae microgreens (broccoli, kale, mustard and radish) was evaluated on colon cancer Caco-2 cells vs. normal colon CCD18-Co cells after 24 h treatment with BFs diluted 1:10 v/v in cell culture medium. Their bioactivity was compared with the digestion blank, while the colon cancer chemotherapeutic drug 5-fluorouracil was used as a positive control. Cell viability (mitochondrial enzyme activity assay (MTT test) and Trypan blue test) and mechanisms related to antiproliferative activity (cell cycle, apoptosis/necrosis, mitochondrial membrane potential, reactive oxygen species (ROS) production, Ca2+ and g…

Addition of milk or caseinophosphopeptides to fruit beverages to improve iron bioavailability?

Abstract A study has been made of the influence of caseinophosphopeptides (CPPs) added to a fruit beverage versus milk based fruit beverages upon iron retention, transport and uptake, using a combined simulated gastrointestinal digestion/Caco-2 cell system. Grape concentrate, orange concentrate, and apricot puree were used for sample formulation. Eight samples were assayed with/without added Fe sulphate (3 mg/100 ml fruit beverage) and/or added Zn sulphate (1.6 mg/100 ml fruit beverage), with/without skimmed milk (11% v/v). The addition of milk to fruit beverages exerted a positive effect on iron retention, transport and uptake versus fruit beverages, and this effect was greater than that o…

Effect of processing and food matrix on calcium and phosphorous bioavailability from milk-based fruit beverages in Caco-2 cells

9 páginas, 4 figuras, 4 tablas.-- et al.

Physiological concentrations of phytosterols enhance the apoptotic effects of 5-fluorouracil in colon cancer cells

Abstract Combining natural products as co-adjuvants in 5-fluorouracil (5-FU) chemotherapy might enhance the effectiveness of 5-FU by avoiding a high dosage and/or reducing treatment times. We explored the anticancer efficacy of the phytosterols (PS) at concentrations achievable in the human colon, as well as their potential as sensitizing agents of human colon cancer cells (Caco-2 and HT-29) to 5-FU treatment. Cells proliferation, combination index, cell cycle, apoptosis, caspases activation, ROS production, and ΔΨm were determined. Co-treatment (PS+5-FU) had an antiproliferative additive effect, and moreover, in general a significantly improved efficacy was observed on cell cycle arrest at…

Selenium, Copper, and Zinc Indices of Nutritional Status : Influence of Sex and Season on Reference Values

The objective was to estimate the possible influence of season or season and sex on the selenium, glutathione peroxidase (GSH-Px), copper, zinc, and superoxide dismutase (SOD) values in plasma with a view to establishing reference values. For this purpose, 55 healthy volunteers (36 women and 19 men) were selected and their whole-blood samples were collected four times a year at the beginning of each season. In the population as a whole, no statistically significant seasonal differences were detected in copper, selenium, and GSH-Px values in plasma, whereas zinc and SOD values in plasma depended on the season. In general, the variability was higher among the women. In view of the differences…

Mathematic predictive models for calculating copper, iron and zinc dialysability in infant formulas

Differences in the dialysability of a mineral element from infant formulas of the same type were detected in a previous study. As these may be due to the effects that different levels of the components of the formulas could have on dialysability, we attempted to establish mathematical models to predict the dialysability of Cu, Fe and Zn from infant formulas according to their ascorbic, citric, or selected amino acid contents. A simple linear regression was applied between the ascorbic acid, citric acid and amino acid contents and the Cu, Fe and Zn dialysability of 18 powdered infant formulas of different types. Significant correlations (P<0.05) were obtained between the dialysability of Fe …

Effect of proteins, phytates, ascorbic acid and citric acid on dialysability of calcium, iron, zinc and copper in soy-based infant formulas.

The possible effect of ascorbic acid, citric acid, proteins and phytate on dialysability of Ca, Fe, Zn and Cu in soy-based infant formulas is studied, taking dialysability as a measure of the amount of element available for absorption. Different dialysis percentages for similar element contents in different formulas are found. A regression analysis was applied between Ca, Zn, Cu and Fe dialysis percentages and soy-based formula components to estimate the possible influence of the latter on the dialysability of the elements. Significant correlations were found between citric acid contents and dialysability of Zn and Fe. No correlations were found between protein, ascorbic acid and phytic aci…

Influence of orange cultivar and mandarin postharvest storage on polyphenols, ascorbic acid and antioxidant activity during gastrointestinal digestion.

Polyphenols, ascorbic acid content and antioxidant activity of two sweet oranges (Navel-N and Cara Cara-CC) and mandarin (Clementine-M) as well as their bioaccessibilities were evaluated in pulps and compared to those in fresh juice. Thus, pulps of oranges and mandarins displayed higher hesperidin (HES), narirutin (NAR), total flavonoids (TF), total phenols (TP) and antioxidant activity (AAC) than their corresponding juices. Also, CC products presented higher bioactive compounds content than N ones. Bioaccessibility of bioactive compounds and AAC were higher in pulps of both oranges and mandarin than in their corresponding juices. Oranges (N and CC) pulps and juices presented higher bioacce…

Effect of a Milk-Based Fruit Beverage Enriched with Plant Sterols and/or Galactooligosaccharides in a Murine Chronic Colitis Model

The potential anti-inflammatory effect of plant sterols (PS) enriched milk-based fruit beverages (PS, 1 g/100 mL) (MfB) with/without galactooligosaccharides (GOS, 2 g/100 mL) (MfB-G) in an experimental mice model of chronic ulcerative colitis was evaluated. Beverages were orally administered to mice every day by gavage to achieve PS and GOS doses of 35 and 90 mg/kg, respectively, and experimental colitis was induced by giving mice drinking water ad libitum containing 2% (w/v) dextran sulphate sodium (DSS) for 7 days, alternating with periods without DSS up to the end of the study (56 days). MfB beverage showed significant reduction of symptoms associated to ulcerative colitis and improved t…

Review: Effect of Some Components of Milk- and Soy-Based Infant Formulas on Mineral Bioavailability

Infants' high nutritional needs are fulfilled by mother's milk or infant formulas to provide all the necessary nutrients, among them minerals. Minerals uptake depends not only on mineral content but also on their bioavailability which, in turn, is affected by the different components of the infant formulas. An understanding of these effects would help to improve mineral bioavailability. This work reviews the influence of endogenous (proteins and phytates) and added (ascorbic and citric acid) components in infant formulas on the bioavailability of nutritionally important mineral elements (calcium, zinc, iron and copper) and their interactions. Special attention is given to the influence of p…

Gangliosides and sialic acid effects upon newborn pathogenic bacteria adhesion: An in vitro study

The effect of the main gangliosides (GM(1), GM(3), GD(3)) and free sialic acid (Neu5Ac) upon the adhesion of pathogenic bacteria implicated in infant diarrhoea is assessed in vitro using the Caco-2 cell line. Concentrations of the bioactive compounds found in the bioaccessible (soluble) fraction of infant formula and human milk are employed. Bacterial adhesion behaviour included enterotoxigenic Escherichia coli (ETEC), enteropathogenic E.coli (EPEC), Listeria monocytogenes, Salmonella entericaserovartyphi, Shigella sonnei, Campylobacter jejuni and Helicobacter pylori. Three different approaches were assayed: pre-incubation of bacteria and compounds before addition to cells (competition); pr…

Anti-Inflammatory and Cytoprotective Effect of Plant Sterol and Galactooligosaccharides-Enriched Beverages in Caco-2 Cells

Plant sterol (PS) (1 g/100 mL) enriched milk-based fruit beverages with or without galactooligosaccharides (GOS) (1.8 g/100 mL) were used in differentiated Caco-2 cells. Their potential cytopreventive effect against oxidative stress induced by cholesterol oxidation products (COPs) and their anti-inflammatory properties were evaluated. Preincubation (24 h) with bioaccessible fractions from beverages without and with GOS (MfB and MfB-G) completely prevented the COPs (60 μM/4 h) induced oxidative stress independent to GOS presence with exception to calcium influx and GSH content, where a partial protective effect was observed. Besides, MfB produced a significant (p < 0.05) reduction of IL-8 (4…

Relative expression of cholesterol transport-related proteins and inflammation markers through the induction of 7-ketosterol-mediated stress in Caco-2 cells.

Human diets contain sterol oxidation products that can induce cytotoxic effects, mainly caused by cholesterol oxides. However, phytosterol oxides effects have been less extensively investigated. This study evaluates the production of inflammatory biomarkers (IL-1β, IL-8, IL-10, TNFα) and the influence of gene expression transporters and enzymes related to cholesterol absorption and metabolism (NPC1L1, ABCG5/8, HMGCoA, ACAT) produced by 7-ketosterols (stigmasterol/cholesterol) in Caco-2 cells. These effects were linked to intracellular signaling pathways by using several inhibitors. Results showed 7-ketostigmasterol to have a greater proinflammatory potential than 7-ketocholesterol. In non-p…

Phospholipids in Human Milk and Infant Formulas: Benefits and Needs for Correct Infant Nutrition.

The composition of human milk has served as a basis for the development of infant formulas, which are used when breastfeeding is not possible. Among the human milk nutrients, 50% of the total energetic value corresponds to fat, with a high level of fatty acids and 0.2-2.0% present in the form of phospholipids (PLs). The PL contents and fatty acid distribution in PL species have been investigated as bioactive elements for the production of infant formulas, since they offer potential benefits for the optimum growth and health of the newborn infant. The differences in the amount of PLs and in fatty acid distribution in PL species between human milk and infant formulas can imply biologically si…

Fortification of milk with calcium: effect on calcium bioavailability and interactions with iron and zinc.

Calcium solubility, dialysability, and transport and uptake (retention + transport) by Caco-2 cells as indicators of calcium bioavailability have been estimated in the in vitro gastrointestinal digests of milk and calcium fortified milk. A significant linear correlation (p < 0.05) was obtained between calcium uptake and the amount of soluble calcium added to the cells, and also between percentage calcium uptake and the calcium measured in the analyzed samples. The solubility, dialysis, transport, and uptake values are higher (p < 0.05) for calcium fortified milks than for nonfortified milks; that is, calcium fortification increases not only calcium content but also its bioavailability. An i…

Bioaccessibility of minerals in school meals: Comparison between dialysis and solubility methods

Abstract Determinations have been made of content and bioaccessibility of Ca, Fe, Zn and Cu in 13 dishes collected from a catering service delivering to a school. Bioaccessibility was estimated by measuring the soluble or dialyzable mineral fraction resulting from in vitro gastrointestinal digestion of the meal. The analyzed dishes had mineral contents (μg/g) in the following ranges: Ca (74.1–913), Fe (2.8–17.9), Zn (2.8–13.1), Cu (0.28–1.90). Mineral solubility and dialysis percentages were as follows: Ca (1.7–96.2; 0.75–61.3), Fe (16.0–97.8; 0.23–19.0), Zn (22.6–93; 5.78–31.45), Cu (35.7–92.3; 0.66–25.0). The highest bioaccessible Ca content corresponded to fish-based dishes, while vegeta…

Gene-diet interaction in plasma lipid response to plant sterols and stanols: A review of clinical trials

Abstract Plant sterols and stanols (PS) are well known for their cholesterol-lowering effect by reducing intestinal absorption of cholesterol. However, genetic factors modulate the low-density lipoprotein cholesterol (LDL-C) response to PS therapy. This review examines clinical trials evaluating the impact of the main genes associated with response of plasma lipid concentrations to PS intake: APOE, CYP7A1, ABCG5/G8, NPC1L1, CETP, APOA4/A5, SCARBI, HMGCR, PPARα, LIPC, MTHFR and LPA. Evidence indicates that carriers of mutant allele of the CYP7A1 c. −204 A > C variant experience a greater plasma cholesterol reduction after PS intake, although there is discrepancy for the rest of genetic varia…

Vitamin E as an IgE inhibitor: stability during cold storage of human milk

Human breast milk is recommended as the unique food for neonates based on its known properties. When the production of milk by themother is not sufcient or the mother is not able to feed her child for professional reasons, milk banks or the mothers’ practice ofcollecting their own milk are the existing alternatives for breast-feeding. In both situations cold storage (refrigeration or freezing) can beused in neonatal units, at home and in human milk banks

Determination of antimony in drinking waters by an inexpensive, reproducible hydride generator for atomic spectroscopy.

A method for determining antimony in drinking waters is described. In order to prevent a substantial error caused by the different oxidation states of antimony, Sb(V) is reduced to Sb(III) with potassium iodide-ascorbic acid. Covalent hydride is generated with a home made device by adding NaBH4. The hydride is then atomized in a flame-heated silica tube and atomic absorption is measured spectrophotometrically. The optimal conditions for this determination are discussed and interference effects are described. Results obtained by determining linearity range (0-200 ng), detection (LOD) and quantitation (LOQ) limits (LOD = 0.347 ng/ml, LOQ = 1.158 ng/ml), precision (instrumental CV 4.08% and me…

Effects of phytosterol ester-enriched low-fat milk on serum lipoprotein profile in mildly hypercholesterolaemic patients are not related to dietary cholesterol or saturated fat intake

Phytosterols (PS) are recommended to reduce LDL-cholesterol. However, the influence of cholesterol and fat intake on the lipid-lowering effect of PS in mildly hypercholesterolaemia is unclear. Thus, the aim of the present study was to evaluate whether the efficacy of PS is related to the composition of saturated fat and dietary cholesterol intake. Additionally, serum carotenoid content was analysed to evaluate to what extent it was undermined by PS. This was a 3-month randomised, parallel trial with a three-arm design. Patients were divided into three groups: healthy diet (n 24), healthy diet + PS (n 31) and free diet + PS (n 29), receiving 2 g/d of PS. Healthy and free diets were character…

In Vitro Dialyzability of Zinc from Different Salts Used in the Supplementation of Infant Formulas

Seven zinc salts--acetate, chloride, lactate, sulfate, citrate, gluconate, and oxide--were added to milk--and soy-based infant formulas to estimate possible differences in zinc availability depending on the type of salt used. For this purpose, an in vitro method that estimates the dialyzability of the element (i.e., the fraction available for absorption) was applied. Zinc dialyzability is always higher in milk-based products than in soy products, even when the total zinc contents are higher in the latter. The salts can be classified according to the zinc dialyzability in the two types of formulas as follows: oxidegluconate = chloride = lactatecitrate = acetatesulfate. Therefore, according t…

An approach to As(III) and As(V) bioavailability studies with Caco-2 cells

Foods and drinking water are the main sources of human exposure to inorganic arsenic [As(III) and As(V)]. After oral ingestion, the intestinal epithelium is the first barrier to absorption of these species. A human intestinal cell line (Caco-2) was used to evaluate cell retention and transport of As(III) (15.6-156.0 microM) and/or As(V) (15.4-170.6 microM). Cell monolayer integrity, cell viability, membrane damage and effects on cell metabolism were evaluated. Only the highest concentrations assayed [As(III): 156.0 microM; As(V): 170.6 microM] produced a cytotoxic effect with different cellular targets: As(III) altered the permeability of tight junctions, and As(V) caused uncoupling of the …

Optimization of selenium determination in human milk and whole blood by flow injection hydride atomic absorption spectrometry.

abstract A flow injection hydride atomic absorption spectrometry (FI-HAAS) method was developed for determining selenium in human milk and whole blood after microwave digestion of the sample. The sample (2 mL human milk or 0.25 mL blood) was introduced into the microwave vessel with 1.5 mL HN03 and 0.25 mL H202 and 300 W (4 min) and 600 W (4 min) were applied. The digestion was completed by heating to 140 C (2-3 h). Se (VI) was reduced to Se (IV) with hydrochloric acid. The instrumental conditions for FI-HAAS (concentrations of reducing agent and carrier acid, flow rate of argon carrier gas, and sample volume injected) were optimized. The detection limit of the proposed method was 0.23 ng/m…

Antiproliferative effects of bioaccessible fractions of honeys from Sicilian black honeybee (Apis mellifera ssp. sicula) on human colorectal carcinoma cells

The aim of this study was to evaluate the antiproliferative activity of bioaccessible fractions (BFs) obtained by the internationally standardized INFOGEST static digestion method to Sicilian honeys of three distinct floral origins (Sulla, Thistle and Dill) and the Manuka honey (gold standard), and to compare their effects based on total polyphenol content (TPC). Differentiated CaCo-2 cells (intestinal-like) and non-differentiated CaCo-2 and HCT-116 colonic tumour-like cells were incubated for 24 h with BFs of honeys to test viability, apoptosis, mitochondrial membrane potential (MMP), ROS and cell cycle. TPC after digestion ranked in the following order: Dill &gt; Thistle &gt; Sulla &gt; M…

Stability of Plant Sterols in Ingredients Used in Functional Foods

The content of plant sterol (PS) and their oxidation products (POPs) in eight ingredients used to enrich functional foods was studied. A gas chromatographic (GC) technique with mass-spectrometric detection was used for identification, while GC with a flame ionization detector (GC-FID) was used for quantification. β-Sitosterol was the most abundant phytosterol, and the main POPs found were derived from this compound (7α/β-hydroxysitosterol, 7-ketositosterol, and sitostanetriol). The total amount of POPs found in the ingredients ranged from 29.03 to 110.02 μg/100 g PS. The β-sitosterol oxidation rates ranged from 10 to 50 μg β-sitosterol oxides/100 g of β-sitosterol. In view of this low rate …

Caseinophosphopeptides exert partial and site-specific cytoprotection against H2O2-induced oxidative stress in Caco-2 cells

Abstract Caseinophosphopeptides can sequester prooxidant metals and scavenge free radicals, and may thus be used as functional food ingredients. The total antioxidant capacity (TEAC and ORAC) of two pools of caseinophosphopeptides (1–3 mg/ml), obtained from casein subjected to simulated gastrointestinal digestion (at two different pH values) and selective precipitation, was evaluated to determine dose–response activity. Pool B (which showed the highest antioxidant capacity due to the presence of more antioxidant amino acids) was used to test its cytoprotective effect against H2O2-induced oxidative stress in Caco-2 cells. Caseinophosphopeptides protected the cells against oxidative damage by…

Plant sterol oxides in functional beverages: Influence of matrix and storage

Three plant sterol (PS)-enriched beverages, milk based fruit juice (MFJPS), fruit juice (FJPS) and milk beverage (MPS), were stored at 4, 24, or 37 °C and analysed at regular time intervals of 2 months until 6 months. PS stability was analysed from the production of phytosterol oxidation products (POPs). The β-sitosterol oxides (7α/7β-hydroxy, β/α-epoxy, triol, and 7-keto) and campesterol oxides (β/α-epoxy, and 7-keto) were detected in all beverages and at all storage times and temperatures. Total POP contents followed the order MPS≫FJPSMFJPS. In general, the beverages showed low PS oxidation levels (0.17%). Predictive models of POP content versus storage time were established. These models…

Effect of caseinophosphopeptides added to fruit beverages upon ferritin synthesis in Caco-2 cells

Ferritin synthesis was evaluated in iron sulphate solutions and in fruit beverages supplemented with iron, with/without zinc in the presence of caseinophosphopeptides (CPPs), based on a system combining simulated gastrointestinal digestion and the Caco-2 cell model. In iron sulphate solutions with/without zinc, the addition of CPPs had no effect upon ferritin synthesis. When CPPs were added to the bioaccessible fraction of fruit beverages supplemented with iron, ferritin synthesis increased in the same proportion as with the addition of milk. The addition of zinc to fruit beverages supplemented with iron and to iron sulphate solutions decreased ferritin synthesis. Nevertheless, in fruit bev…

HPLC Method for Cyst(e)ine and Methionine in Infant Formulas

Cyst(e)ine and methionine were converted into cysteic acid and methionine sulfone by oxidizing with performic acid. The oxidized samples were then subjected to acid hydrolysis (6N HCl, 105-110°C/24 hr). After derivatization with phenylisothiocyanate, reverse phase HPLC separation was carried out at 48°C and with UV detection. Different gradients and pH values in the eluent were assayed to determine the best resolution. Analytical parameters, detection and quantification limits, linearity, precision and accuracy, were determined. The method was reliable and accurate for measuring cyst(e)ine and methionine in infant formulae.

Lipid peroxidation and antioxidant enzyme activities in patients with type 1 diabetes mellitus

To evaluate oxidative stress in type I diabetes mellitus, two antioxidant enzymes in erythrocytes, copper-zinc superoxide dismutase (SOD EC 1.15.1.1.) and seleno-dependent glutathione peroxidase (GSH-Px; EC 1.11.19), and two indexes of peroxidation in plasma, thiobarbituric acid reactive substances (TBARS) and organic hydroperoxides (OHP), were measured in 118 patients with insulin-dependent diabetes mellitus (IDDM), classified in accordance with the presence or absence of vascular complications and the degree of metabolic control established by the HbA1c level. Ninety healthy subjects made up the control group. According to our results, plasmatic TBARS and OHP concentrations are significan…

Anti-proliferative effect of main dietary phytosterols and β-cryptoxanthin alone or combined in human colon cancer Caco-2 cells through cytosolic Ca+2 – and oxidative stress induced apoptosis

β-cryptoxanthin (β-Cx) and phytosterols (Ps) have potential against different cancer types, including colon cancer. However, their combined action has not been reported so far. Human colon cancer Caco-2 cells were treated 24 h with β-Cx and/or main dietary Ps (β-sitosterol, campesterol and stigmasterol), alone or in combination, at concentrations compatible with physiological human serum levels. A decrease in cell viability due to apoptosis (rise in sub-G1 population and exposure of membrane phosphatidylserine) was accompanied with dephosphorylation of BAD, mitochondrial depolarization and caspase 3-dependent PARP cleavage, with intracellular Ca2+ influx and increase of RONS levels as initi…

Mercury and selenium in fish and shellfish: Occurrence, bioaccessibility and uptake by Caco-2 cells

This study evaluates Hg and Se concentrations and bioaccessibility (element solubilised after simulated gastrointestinal digestion) in 16 raw seafood species consumed in Spain. The concentrations varied greatly (Hg, 3.8-1621 ng/g wet weight, ww; Se, 84-1817 ng/g ww). Only one sample of swordfish exceeded the Hg limit permitted in Spain (1mg/kg), and for this sample the Hg/Se molar ratio and Se Health Benefit Value food safety criteria also indicated the presence of a risk. Bioaccessibility of Hg (35-106%) and Se (17-125%) was very variable and the Hg/Se molar ratio in the bioaccessible fraction was less than one for all samples. Transport by Caco-2 cells, an intestinal epithelium model, was…

Bioavailability of calcium from milk-based formulas and fruit juices containing milk and cereals estimated by in vitro methods (solubility, dialyzability, and uptake and transport by caco-2 cells).

An adequate calcium intake during the first years of life is needed for normal growth and development and to prevent rickets. The bioavailability of calcium from infant foods (milk-based formulas and fruit juices containing milk and cereals, FMC), the dietary sources of calcium in these stages of life, has been estimated on the basis of simulated gastrointestinal digestion and calcium solubility and dialyzability values and on the efficiency of transport and uptake by Caco-2 cells. The ranking of samples according to calcium bioavailability depends on the use of solubility or dialyzability as criterion. On the basis of the former, the highest value corresponded to adapted formulas and the l…

Selenium, zinc and copper in plasma of patients with type 1 diabetes mellitus in different metabolic control states

The Studies of selenium (Se), zinc (Zn) and copper (Cu) levels in diabetic patients have led to contradictory findings as the possible relationship between the degree of diabetic control and the changes in mineral contents. In the present study the plasma Cu, Se, and Zn contents of diabetic patients and healthy people were measured and the relationship between these contents and diabetic metabolic control, as determined by glycosylated hemoglobin (HbA1c), was studied. The mean plasma Se content in diabetic patients was significantly lower than in controls (p < 0.01) and a negative correlation between the plasma contents of Se and HbA1c was found. No statistically significant differences in …

Calcium bioavailability in human milk, cow milk and infant formulas—comparison between dialysis and solubility methods

Abstract The percentages of total, soluble and dialysable calcium of human milk, cow milk and milk and soy based infant formulas were determined in order to detect possible differences in the calcium bioavailability of the samples. For this purpose an in vitro method was applied to these four calcium sources. The ranking of the analysed samples in terms of calcium bioavailability depends on the criteria applied. Calcium ranked dialysis percentage was: cow milk>human milk>soy based formula>milk based formulas. Calcium ranked solubility percentage was: human milk>cow milk>soy-based formula>milk-based formulas. Comparison of the results of the in vitro assay with the information available on i…

Effect of β-cryptoxanthin plus phytosterols on cardiovascular risk and bone turnover markers in post-menopausal women: a randomized crossover trial.

Abstract Background and aim Post-menopausal women are at higher risk of cardiovascular disease and bone demineralization. Phytosterols (PS) may be used for hypercholesterolemia in some groups and β-cryptoxanthin (β-Cx) displays a unique anabolic effect on bone. Our aim was to assess the changes in cardiovascular and bone turnover markers from the oral intake of β-Cx and PS in post-menopausal women. Methods and results A randomized, double-blind, crossover study with β-Cx (0.75 mg/day) and PS (1.5 g/day), single and combined, was performed in 38 postmenopausal women. Diet was supplemented with 1 × 250 mL milk-based fruit drink/day for 4 weeks with a wash-out period of 4-weeks in between. Ser…

Dialysability of Calcium, Iron, and Zinc in Beans, Chick Peas, and Lentils

As2O3-induced oxidative stress and cycle progression in a human intestinal epithelial cell line (Caco-2)

Foods and drinking water are the main routes for human exposure to inorganic arsenic, the intestinal epithelium being the first barrier against such exogenous toxicants. The present study evaluates the effect of As(III) (0.5-25 microM) upon Caco-2 cells as an intestinal epithelia model. Cell viability, intracellular formation of reactive oxygen species (ROS), mitochondrial membrane potential (Deltapsim) changes, and cell cycle distribution in exposed cultures were evaluated. The intracellular production of ROS was seen to increase in a non-dose dependent manner at all concentrations tested, with impairment of cell mitochondrial enzyme function secondary to a loss of Deltapsim. Concentration…

Simultaneous quantification of serum phytosterols and cholesterol precursors using a simple gas chromatographic method

Determination of the main phytosterols (Ps, β-sitosterol and campesterol) and cholesterol precursors (desmosterol and lathosterol) in human serum using a simple GC-FID method has been validated. Direct saponification, without lipid extraction, sterols extraction, and further derivatization was applied to samples prior to GC analysis. To evaluate the method, a pool of serum samples from eight healthy women was used. Good linearity (r>0.99) was found in the assay range: β-sitosterol (0.99–17.82 µg/mL), campesterol (0.14–10.8 µg/mL), desmosterol (0.17–2.6 µg/mL), and lathosterol (0.6–5.97 µg/mL). Limits of detection (ng/mL) were: 86 (β-sitosterol), 42 (campesterol), 4 (desmosterol), and 44 (la…

Effect of processing on the bioaccessibility of bioactive compounds – A review focusing on carotenoids, minerals, ascorbic acid, tocopherols and polyphenols

Health benefits of bioactive compounds depend not only on the intake levels but also on their bioavailability (BAv). In vitro methods to simulate gastro-intestinal digestion allow to determine the bioaccessibility (BAcs) of these compounds, as a first step of BAv, and can be used to evaluate the effect of processing on them to design functional foods with increased health-promoting effects. The impact of traditional processing technologies such as thermal treatment and novel emerging non-thermal technologies such as high pressure processing, high-intensity pulsed electric fields and ultrasound on BAcs of bioactive compounds as carotenoids, minerals, ascorbic acid, tocopherols, polyphenols a…

Current methodologies for phytosterol analysis in foods

Abstract Plant sterols (PS) (phytosterols and phytostanols) are bioactive compounds of all vegetable foods where can be found as free sterol alcohols and as conjugates. These latter forms have been less studied, although they may have potential beneficial effects, whereas some PS have several approved health claims, especially in lowering blood cholesterol levels. In this context, it is necessary to establish analytical methods for food authentication, assessing PS health benefits and unfolding future applications. Several extraction methodologies have been employed for the extraction of PS from food matrixes (usually solvent extraction and saponification) although solid-phase extraction, s…

Calcium dialysability as an estimation of bioavailability in human milk, cow milk and infant formulas

Abstract The calcium dialysability of human milk, cow milk and infant formulas is estimated by applying the in vitro method (Miller, Schricker, Rasmussen, & Van Campen, (1981). American Journal of Clinical Nutrition, 34, 2248–2256) slightly modified, as a measure of its bioavailability. The influence of the protein composition of infant formula samples on calcium bioavailability is also studied. Whole cow milk, a pool of human milk and 18 commercial infant formulas of 6 different types are analysed. The highest values of calcium dialysability (mg Ca/100 ml infant formula) correspond to premature (13.6) and follow-up formulas (11.8) and the lowest ones to adapted formulas (3.7); the latter v…

Impact of processing on mineral bioaccessibility/bioavailability

Abstract A review is made of the influence of processing on the bioaccessibility/bioavailability of macrominerals (Ca, P, Mg and K) and microminerals (Fe, Cu, Zn and Se). In vegetable foods, household processes (dehulling, soaking, germination and fermentation) alone or with thermal processing increase mineral bioaccessibility, mainly referred to Fe, Cu, Zn, Se and Ca. The application of thermal processing to vegetable foods has different effects upon mineral bioaccessibility, with an overall increase in Ca and Fe and a decrease in Se. In turn, there is a general positive impact upon mineral bioaccessibility in animal foods when conventional thermal processing is used with shorter times and…

In vitro bioavailability of iron and calcium in cereals and derivatives: A review

Cereals are a staple food in both developed and developing countries, and are considered to be the best vehicle for iron and calcium fortification, as an important strategy for combating dietary deficits. Inadequate dietary intake of iron and calcium is related to a number of disease conditions such as anemia, osteoporosis, hypertension, and different cancers. From a nutritional point of view, it is interesting to know not only the amount of minerals consumed, but also their bioavailability. The present study reviews the current knowledge on the in vitro bioavailability of iron and calcium in cereals, placing emphasis on the methodologies used and on the influence of dietary factors and foo…

Hypercholesterolemic patients have higher eryptosis and erythrocyte adhesion to human endothelium independently of statin therapy

BACKGROUND Phosphatidylserine (PS) externalization out of the membrane facilitates the eryptotic erythrocytes (EE) binding to endothelial cells (EC), potentially leading to atherosclerosis. Thus, the levels of eryptosis and interactions of EE-EC in hypercholesterolemic patients, either non-medicated or medicated, compared with healthy subjects were studied. METHODS A total of 56 subjects clustered into three groups: (control (n = 20), hypercholesterolemic non-treated (HCNT) (n = 15), and statin-treated (HCT) (n = 21)) were enrolled in this cross-sectional study. Biochemical parameters were determined with validated and standard methods. PS exposure was estimated from annexin-V-binding, cell…

Extending in vitro digestion models to specific human populations: Perspectives, practical tools and bio-relevant information

29th International Conference of the European-Federation-of-Food-Science-and-Technology (EFFoST) -- NOV 10-12, 2015 -- Athens, GREECE

Cobalt content of foods and diets in a Spanish population

The WHO Expert Committee on Food Contaminants has pointed out the need of determining the content of foods in trace elements in order to ascertain their contribution to the total intake of oligoelements through the diet. Since the contents in trace elements depend on the environmental features, it is necessary to carry out systematic determinations of trace elements in foods of local origin in order to know the exposure level of population. This work intends to study the cobalt contribution through the diet. The element intake is calculated from the cobalt content in raw foods after elimination of the non-eatable moiety by flame atomic absorption spectroscopy, after formation of a chelate w…

Iron Bioavailability in Fortified Fruit Beverages Using Ferritin Synthesis by Caco-2 Cells

The bioavailability of iron from fortified fruit beverages was estimated by an in vitro system including enzymatic digestion, iron uptake by Caco-2 cells, and ferritin formation determined via an enzyme immunoassay (ELISA). Thus, the aim of the present study was to assess iron bioavailability as influenced by the presence of known dietary promoter and inhibitory factors in fortified fruit beverages containing iron and/or zinc and/or skimmed milk. No negative effect (p > 0.05) derived from micronutrient interaction can be ascribed to zinc supplementation on iron availability. Besides, the presence of caseinophosphopeptides derived from casein hydrolysis during digestion may confer enhancing …

Bioaccessibility of tocopherols, carotenoids, and ascorbic acid from milk- and soy-based fruit beverages: Influence of food matrix and processing

A study was made of the effect of high-pressure processing (HPP) and thermal treatment (TT) on plant bioactive compounds (tocopherols, carotenoids, and ascorbic acid) in 12 fruit juice-milk beverages and of how the food matrix [whole milk (JW), skimmed milk (JS), and soy milk (JSy)] modulates their bioaccessibility (%). HPP (400 MPa/40 °C/5 min) produced a significant decrease in carotenoid and ascorbic acid bioaccessibility in all three beverages and maintained the bioaccessibility of tocopherols in JW and JS while decreasing it in JSy. TT (90 °C/30 s) produced a significant decrease in tocopherol and carotenoid bioaccessibility in all three beverages and increased the bioaccessibility of …

Review: Determination of Vitamin D in Dairy Products by High Performance Liquid Chromatography

This work reviews the methods used for the determination of vitamin D in some dairy products (milk and infant formulas) by high performance liquid chromatography (HPLC). The low vitamin D contents and the presence of interfering compounds require sample treatment and purification of the extract. The advantages and drawbacks of hot and cold saponification, direct extraction and different types of extract purification are also discussed, taking into account the lack of vitamin D stability by heating, exposure to light and oxidation. With respect to chromatographic determination, the mode (normal or reverse phase), type of column (microcolumn or conventional) and detection system (UV, electro…

Plant Sterols and Antioxidant Parameters in Enriched Beverages: Storage Stability

Plant sterols (PS) stability, antioxidant parameters, and color were studied during 6 months of storage at 4, 24, and 37 °C in three PS-enriched functional beverages. Beverages were skimmed milk with fruit juice and PS (MFJPS), fruit juice and PS (FJPS), and skimmed milk with PS (MPS). No loss in total PS content occurred during storage observing the same values at any given storage time point. Total carotenoids decreased 36% with storage at two months and then remained stable. Total polyphenols showed fluctuations throughout the storage, remaining stable at 6 months and reaching initial values. The antioxidant capacity (TEAC method) increased 18% at 6 months, and there was an increase in c…

Calcium, Iron, and Zinc Uptake from Digests of Infant Formulas by Caco-2 Cells

Our aim was to estimate the bioavailability of calcium, iron, and zinc from infant formulas using a model that includes in vitro digestion and a Caco-2 cell culture to estimate the uptake. The cell culture conditions were selected, and uptake assays were carried out first with calcium, iron, and zinc standard solutions, and then with the soluble fraction of enzymatic digests of an adapted milk-based and a soy-based infant formula. It was not possible to measure the uptake of calcium, iron, and zinc from standard solutions added to the cell cultures in amounts similar to those present in infant formula digests with our method. The fact that it was, however, possible in the case of enzymatic …

Effect of Caseinophosphopeptides from αs- and β-Casein on Iron Bioavailability in HuH7 Cells

International audience; Two pools of caseinophosphopeptides (CPPs) obtained from αs- and β-casein fractions (α-CPPs and β-CPPs) were characterized. A total of 16 CPPs were identified in the α-CPPs pool, 9 of them derived from αs1-casein and 7 from αs2-casein. A total of 18 CPPs were identified in the β-CPPs pool. Four of the identified CPPs contained the characteristic phosphoseryl-glutamic acid cluster SpSpSpEE. Calcein assay was used to compare the iron-binding capacity of the α- and β-CPPs pools. At the concentration of 12.5 μM CPPs used in the iron bioavailability assays, β-CPPs pools show greater iron-binding capacity than α-CPPs pools. HuH7 human hepatoma cells show many differentiate…

Sialic acid (N-acetyl and N-glycolylneuraminic acid) and ganglioside in whey protein concentrates and infant formulae

Abstract Sialic acid and gangliosides content and profiles were analyzed in infant formulae, whey protein concentrates and human milk. In infant formulae, N-acetylneuraminic (Neu5Ac) and N-glycolylneuraminic (Neu5Gc) acid ranged from 147.6–199.7 to 3.3–8.3 mg L −1 , in whey protein concentrate from 1.6–2.4 g 100 g −1 to 2.8–20.2 mg 100 g −1 and in human milk from 299.9 to 2.1 mg L −1 . Gangliosides ranged from 0.25 to 2.29 mg lipid-bound sialic acid L −1 in reconstituted infant formula, the main gangliosides being GD 3 (73.3–55.5%), GM 3 (3.5–36.6%) and GT 3 (1.4–14.1%) and O-acetyl-GD 3 (5.3–18.9%). In whey protein concentrates, 0.7–55.6 mg lipid-bound sialic acid 100 g −1 were found with …

Selenium contents of human milk and infant formulas in Spain.

The selenium content of Spanish human milk samples and different milk-based and soy-based infant formulas has been estimated by using a flow injection hydride atomic absorption spectrometric method after microwave digestion of the organic matter. Mean values of 11.4 +/- 3.7 and 10.7 +/- 4.6 ng/ml for colostrum and transitional milk, 8.4 +/- 3.4 and 5.3 +/- 1.9 ng/ml for mature milk at 1 month and up to 2 months respectively, was obtained. These values are close to those reported by others authors in Europe, and lower than the ones from the US, Japan and Korea. Selenium contents of the analyzed infants' formulas ranged from 2.7 to 9.6 ng/ml and from 1.8 to 7.5 ng/ml for soy and milk-based in…

Carotenoid bioaccessibility in pulp and fresh juice from carotenoid-rich sweet oranges and mandarins.

Citrus fruits are a good source of carotenoids for the human diet; however, comparative studies of carotenoids in different citrus food matrices are scarce. In this work the concentration and bioaccessibility of carotenoids in sweet oranges and mandarins with marked differences in carotenoid composition were evaluated in pulp and compared to those in fresh juice. The pulp and juice of the red-fleshed Cara Cara sweet orange variety was highly rich in carotenes (mainly lycopene and phytoene) compared to standard Navel orange, while β-cryptoxanthin and phytoene predominated in mandarins. Total carotenoid content in the pulp of the ordinary Navel orange and in the red-fleshed Cara Cara orange, …

Plant sterols from foods in inflammation and risk of cardiovascular disease: a real threat?

High dietary intakes of cholesterol together with sedentary habits have been identified as major contributors to atherosclerosis. The latter has long been considered a cholesterol storage disease; however, today atherosclerosis is considered a more complex disease in which both innate and adaptive immune-inflammatory mechanisms as well as bacteria play a major role, in addition to interactions between the arterial wall and blood components. This scenario has promoted nutritional recommendations to enrich different type of foods with plant sterols (PS) because of their cholesterol-lowering effects. In addition to cholesterol, PS can also be oxidized during food processing or storage, and the…

Evaluation of Antimony, Cadmium and Lead Levels in Vegetables, Drinking and Raw Water from Different Agricultural Areas

Abstract Lead and cadmium levels in edible vegetables and antimony, lead and cadmium in drinking and raw waters from three agricultural areas exposed to different levels of environmental pollution (1-high industrial pollution, 2-high urban pollution, 3-standard low industrial and urban pollution) are determined. The organic matter is destroyed by repeated attack with nitric acid. Cadmium and lead are determined by graphite furnace atomic absorption spectrometry (GFAAS) and antimony by hydride generation atomic absorption spectrometry (HGAAS). When the results obtained for three different areas are compared, differences between cadmium and lead contents in vegetables are observed. Waters are…

Safe intake of a plant sterol-enriched beverage with milk fat globule membrane: Bioaccessibility of sterol oxides during storage

Abstract Sterols in foods are susceptible to oxidation to form oxysterols. It is interesting for consumer health to know real intake and the possible associated adverse effects associated to oxysterols. This study measured oxysterol formation and bioaccessibility (BA) in a plant sterol (PS)-enriched milk-based fruit beverage with milk fat globule membrane (MFGM) added at 0, 3 and 6 months of storage at room temperature. The same cholesterol (COPs) and phytosterol oxidation products (POPs) (exclusively from β-sitosterol) (7α/β-hydroxy, α/β-epoxy, triol and 7-keto) were detected in the beverage and its bioaccessible fraction. Total COPs and POPs contents were maintained during storage, and th…

Iron and zinc bioavailability in Caco-2 cells: Influence of caseinophosphopeptides

Abstract A study has been made of the influence of two pools of caseinophosphopeptides (CPPs) obtained from α s - and β-casein (CN) fractions, and of three specific CPPs (β-CN(1–25)4P, α s1 -CN(64–74)4P and α s2 -CN(1–19)4P), on iron bioavailability (ferritin synthesis) and zinc bioavailability (retention, transport and uptake of zinc) in Caco-2 cells. α-CPP and β-CPP pools did not improve ferritin synthesis, but the three specific CPPs showed an increase in ferritin synthesis in Caco-2 cells versus iron sulphate, β-CN(1–25)4P being the most effective. In relation to zinc bioavailability, α-CPPs, β-CPPs, α s1 -CN(64–74)4P and β-CN(1–25)4P increased zinc uptake. However, this increase was of…

Liquid chromatographic determination of Vitamin D3 in infant formulas and fortified milk

Abstract An isocratic reverse-phase liquid chromatographic (LC) method with electrochemical (EC) detection has been developed and validated for determining Vitamin D 3 in fortified milk and infant formulas. LC–EC determination provides linear responses in the range from 0.03 to 0.7 μg Vitamin D 3 /ml, with inter- and intra-day variations (R.S.D.%) of 4.1 and 4.4, respectively, and detection and quantification limits of 0.012 ng in assay (48 ng/100 g of sample) and 0.039 ng in assay (156 ng/100 g of sample), respectively. Application of the method to a set of infant formulas and fortified milk confirmed its usefulness.

Casein phosphopeptides released by simulated gastrointestinal digestion of infant formulas and their potential role in mineral binding

Abstract Adapted and follow-up milk-based infant formulas were subjected to gastrointestinal digestion simulating physiological conditions. The naturally occurring casein phosphopeptides (CPPs) generated were fractionated by anion exchange high-performance liquid chromatography and sequenced by tandem mass spectrometry. In both infant formula digests, a total of 19 CCPs from bovine casein were identified, of which 7 corresponded to α s1 -casein and 12 to α s2 -casein. Most CPPs had the cluster sequence SpSpSpEE, representing the binding sites for minerals. The distribution of calcium, iron and zinc content in CPP fractions eluted from the anion exchange column was also studied. The results …

Antioxidant effect of casein phosphopeptides compared with fruit beverages supplemented with skimmed milk against H2O2-induced oxidative stress in Caco-2 cells

Abstract Casein phosphopeptides (CPPs) have been proposed as potential dietary antioxidants on designing new functional products. A fruit beverage, with/without milk, was subjected to in vitro digestion. Caco-2 cultures were preincubated with soluble fractions or isolated CPPs, the latter obtained from skimmed milk. The mitochondrial activities (MTT test), intracellular GSH and GSH-reductase activity (GSH-Rd), cell cycle analysis and RNA distribution in cycle phases were studied after inducing oxidative stress status (5 mM H2O2). MTT conversion was better preserved by soluble fractions of fruit beverages, either with or without milk. GSH concentration was equally decreased, and GSH-Rd was i…

Protective effect of bioaccessible fractions of citrus fruit pulps against H2O2-induced oxidative stress in Caco-2 cells

Fruit pulps from Navel (N) and Cara Cara (CC) oranges, and Clementine mandarin freshly harvested (M) and refrigerated stored (M12) were used to evaluate the cytoprotective effect of their bioaccessible fractions (BF) against H2O2-induced oxidative stress in Caco-2 cells. BF of samples preserved viability vs. H2O2 treated cells, reaching values similar to controls. Lipid peroxidation was reduced to levels of control cells, but M did not reach control values. ROS and mitochondrial membrane potential changes (Δψm) values were reduced compared with H2O2 treated cells, but without achieving control levels. A significant reduction in cell proportions in G1 phase and a significant increase in sub-…

Estimation of Arsenic Bioaccessibility in Edible Seaweed by an in Vitro Digestion Method

The aim of this study was to examine the bioaccessibility (maximum soluble concentration in gastrointestinal medium) of total (AsT) and inorganic (AsI) arsenic contents and the effect on them of cooking edible seaweed, a food of great interest because of its high As content. An in vitro gastrointestinal digestion (pepsin, pH 2, and pancreatin−bile extract, pH 7) was applied to obtain the mineral soluble fraction of three seaweeds (Hizikia fusiforme, Porphyra sp., and Enteromorpha sp.). AsT was determined by dry-ashing flow injection hydride generation atomic absorption spectrometry. AsI was determined by acid digestion, solvent extraction, and flow injection hydride generation atomic absorp…

Effectiveness of microwave based digestion procedures for the demineralization of human milk and infant formulas prior to fluorometric determination of selenium

The use of two types of microwave oven, one domestic (A) and the other specially designed for laboratory (B) for microwave-based digestion, to destroy the organic matter in milk and infant formulas prior to fluorometric determination of selenium is studied in order to check their applicability and reliability. In both systems the best results were obtained using the acid mixture HNO3—H2O2, and additional treatment with perchloric acid (60%) in a Thermoblock. The application of two different microwave-based digestion procedures to infant formulas was free of interferences. In the case of human milk, it was necessary to use the standard addition method. Values obtained for detection and quant…

Availability of polyphenols in fruit beverages subjected to in vitro gastrointestinal digestion and their effects on proliferation, cell-cycle and apoptosis in human colon cancer Caco-2 cells

Abstract Three fruit beverages: Fb (grape–orange–apricot), FbFe (Fb + iron sulphate) and FbFeM (FbFe + skimmed milk) were subjected to in vitro gastrointestinal digestion. Addition of Fe and milk decreased the phenolic content of Fb by 13% and 55%, respectively. The digestion process decreased the phenolic content by 47%, 60%, and 70% in Fb, FbFe and FbFeM with respect to the initial, non-digested, Fb. Caco-2 cells were incubated for 4 h daily for four days or continuously for 24 h with bioaccessible fractions obtained after the digestion (digests). Polyphenols were not metabolised by Caco-2 cells. Fb digest (∼50 μM total phenolics) was the sample that led to the highest inhibition of cell …

Relationship between cobalt, copper and zinc content of soils and vegetables

The relationship between cobalt, copper and zinc content of soils and the vegetables grown in them is discussed. Samples of soil and edible vegetables were taken from 15 different sites, corresponding to four agricultural areas exposed to different degrees of environmental pollution: high industrial and traffic, high industrial and urban, and low industrial and urban pollution. Elements were determined by atomic absorption spectroscopy. Variance analysis (ANOVA) was applied to cobalt, copper and zinc content of vegetables and soils in order to detect significant differences among soils of different areas and the vegetables grown there. The possible relationship between the heavy metal conte…

Determination of Cd, Co, Cu, Fe, Pb, Mn, Ni and Zn in diets: Development of a method

A method useful for the determination of cadmium, cobalt, copper, iron, lead, manganese, nickel and zinc in diets is described. Organic matter is destroyed applying a wet procedure, and element content is measured by flame atomic absorption spectroscopy (FAAS) in the case of copper, iron, manganese, nickel and zinc, and by graphite furnace (GF-AAS) in the case of cadmium, cobalt and lead. The matrix interference study is carried out. Values found for linearity, detection and quantitation limits as well as accuracy show that the described method is useful to determine the elements taken into account at usual levels in diets.

Ferritin synthesis by Caco-2 cells as an indicator of iron bioavailability: Application to milk-based infant formulas

The bioavailability of iron from milk-based infant formulas was estimated by an in vitro system including enzymatic digestion, iron uptake by Caco-2 cells and ferritin determination via an enzymoimmunoassay (ELISA). Positive correlations (p < 0.01) were found between the Fe(II) added to Caco-2 cells and ferritin synthesis and between the amount of dialyzed iron added to the cell culture and ferritin synthesis. The comparison of the bioavailability of iron from different milk-based formulas showed that adapted formulas having the same composition but differing in the iron salts added yielded similar ferritin levels. The same happened with follow-up formulas differing only in the presence or …

Evaluation of the cytotoxic effect of 7keto-stigmasterol and 7keto-cholesterol in human intestinal (Caco-2) cells

The biological implications of cholesterol oxidation products have been investigated, though research on plant sterol oxidation products is scarce and in some cases contradictory. The cytotoxicity of 7keto(k)-stigmasterol versus 7keto(k)-cholesterol at different concentrations (0-120 μM) and incubation times (4-24h), in intestinal epithelial cells (Caco-2 cells) was evaluated. The 3-[4,5-dimethylthiazol-2-yl]-2,3-diphenyl tetrazolium bromide and neutral red uptake tests, mitochondrial membrane potential (ΔΨm), and relative DNA and RNA contents in the cell cycle phases were determined. Possible interaction effects between 7k-derivatives or non-oxidized stigmasterol were monitored. Endo/lysos…

Bioavailability of inorganic arsenic in cooked rice: practical aspects for human health risk assessments.

Arsenic is present in rice grain mainly as inorganic arsenic. Little is known about the effect of cooking on inorganic arsenic content in rice and its bioavailability. This study evaluated total arsenic and inorganic arsenic in rice cooked with arsenic-contaminated water, the bioaccessibility of As(III) and As(V) after simulated gastrointestinal digestion, and the extent of arsenic retention and transport by Caco-2 cells used as a model of intestinal epithelia. After cooking, inorganic arsenic contents increase significantly. After simulated gastrointestinal digestion, the bioaccessibility of inorganic arsenic reached 63-99%; As(V) was the main species found. In Caco-2 cells, arsenic retent…

Determination of cobalt in foods by atomic absorption and inductively coupled plasma spectrometry (Short communication)

Effect of Enzyme Amounts Used in Gastrointestinal Digestion Upon Solubility and Caco-2 Cell Uptake Assays of Minerals from Infant Formulas

The effect of enzyme amounts used in gastrointestinal in vitro digestion upon the solubility and Caco-2 cell uptake of calcium, iron and zinc from infant formulas (IFs) was studied. Different amounts of enzymes (g enzyme/g IF), pepsin (0.002 and 0.048), pancreatin (0.0005, 0.002 and 0.01) and bile extract (0.003, 0.125 and 0.0625) were assayed. Mineral soluble contents and mineral uptakes by Caco-2 cells were affected by the enzyme amounts used in digestion. Although the highest mineral solubility (Ca 98.6 vs 46.2%; Fe 98.1 vs 83.9%; Zn 98.4 vs 83%) was obtained when the lowest enzyme (pepsin 0.002 vs 0.048; pancreatin 0.0005 vs 0.01g/g IF) and bile extract (0.003 vs 0.0625g/g IF) amounts …

Effect of cooking and legume species upon calcium, iron and zinc uptake by Caco-2 cells

Abstract An in vitro system, consisting of simulated gastrointestinal digestion and Caco-2 cell culture, was used to estimate the uptake of calcium, iron and zinc from white beans, chickpeas and lentils, and the effect of cooking upon uptake, with the ultimate aim of evaluating legumes as a dietary source of the aforementioned minerals. In raw products, differences were observed in the uptake percentages by Caco-2 cells of a same mineral from different legumes, although these were not related to the total mineral content. In the three elements studied, the highest uptake values corresponded to chickpeas. Traditional cooking significantly ( p 0.05 ) increased the uptake (%) of calcium, iron …