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Bioaccessibility of plant sterols (PS) in an enriched wholemeal rye bread was evaluated, for the first time, using the INFOGEST protocol without gastric lipase (GL) and cholesterol esterase (CE), with GL or GL + CE. Moreover, human chewing and an

The prevention of cardiovascular disease by fortified foods with plant sterols (PS), with a hypocholesterolemic effect, is important for the elderly population. This study aimed to identify the different PS...

Bread is one of the staple foods of many countries, and its enrichment with bioactive compounds is trending. This phenomenon is focused on breads with a good nutritional profile, such as wholemeal rye bread (WRB), in which enrichment with plant sterols (PSs) is allowed in accordance with European regulations. The objective of the present study was to optimize the production of a WRB enriched with PS (PS-WRB) and to evaluate the proximate composition and starch digestibility as an indicator of nutritional quality. The rheological analysis showed that the bread dough presents satisfactory farinographic properties (dough development time 6 min; stability 4 min; degree of softening 100 Brabender units) but high water absorption (67%). The PS-WRB is high in dietary fiber and low in protein (20.4 and 7.7% w/w, dry basis, respectively) compared with other cereals reported in the scientific literature. In turn, a low starch proportion was hydrolyzed during the simulated digestion (59.9% of total starch), being also slowly hydrolyzed, as deduced from the rapidly digestible starch value (56.5% of total starch). In conclusion, WRB is a suitable matrix for PS enrichment, which allows for obtaining a product with a good nutritional profile and potential health benefits., This publication is part of the projects PID2019-104167RB-I00 and PID2019-107650RB-C21 funded by MCIN/AEI/10.13039/501100011033. Mussa Makran holds an FPU19/00156 grant from the Ministry of Universities (Spain).

Elderly people suffer from a higher cardiovascular risk. Thus, the fortification of foods with plant sterols (PSs), which have a cholesterol-lowering function, could be of great interest for this target group. To date, no studies have analyzed how the gastrointestinal conditions of the elderly affect PS bioaccessibility. Therefore, this study evaluated the impact of the adaptation of the gastric phase alone and in combination with the intestinal phase on sterol bioaccessibility. For this purpose, the standardized INFOGEST 2.0 method previously adapted for sterol bioaccessibility evaluation in healthy adults was applied to PS-enriched milk-based fruit beverages, examining changes in enzyme activity, incubation time, agitation and pH, based on elderly physiology. The results suggest that the specific gastrointestinal conditions of the elderly could increase absorption of PSs, since their bioaccessibility (%) in a PS-enriched milk-based fruit beverage was significantly increased compared with that in adults (14.95 ± 0.33

In the present study, the impact of galactooligosaccharide (GOS) addition to a plant sterol (PS)-enriched beverage on the hypocholesterolemic effect and on the bioavailability and colonic metabolization of sterols was evaluated. A crossover trial was undertaken in postmenopausal women who intook a PS-enriched (2 g PS/day) or PS-GOS-enriched beverage (2 g PS/day and 4.3 g GOS/day) for 6 weeks. The presence of GOS did not modify the hypocholesterolemic effect of the PS-enriched beverage (total- and low-density lipoprotein-cholesterol reductions) or sterol bioavailability (increments of serum markers of dietary PS intake and of cholesterol synthesis). The consumption of both beverages led to an increase of sterol and metabolite excretion (with the exception of coprostanol, which decreased) and to slight changes in women's capacities for sterol conversion, regardless of the GOS presence. This study demonstrates the suitability of simultaneous enrichment with PS and GOS in milk-based fruit beverages, considering their hypocholesterolemic effect.

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 volume from forward scatter (FSC), and GSH from CMFDA fluorescence by flow cytometry. The erythrocyte-EC adhesion assay was performed by using the parallel-plate flow chamber technique. RESULTS Higher PS externalization and adhesion of erythrocytes to EC (P

This study evaluates the influence of increasing bile salts and the addition of key enzymes of the lipidic metabolism in the INFOGEST digestion method on sterol bioaccessibility from a plant sterol (PS)-enriched beverage. The assayed modifications were increasing concentration of bovine bile salts (10 vs. 17.5 mM), and addition of gastric lipase (GL) (60U/mL), cholesterol esterase (CE) (0.075 or 2U/mL) or both. Compared to the original method (10 mM bile salts without enzymes), the assayed conditions significantly reduced bioaccessibility of individual (from 11.3 to 19.7 to 5.1-16.6%) and total PS (13.7 to 6.9-8.0%), and cholesterol (52.8 to 20.9-26.1%), except only when CE is added not allowing cholesterol quantification. The bioaccessibility achieved when lipolytic enzymes were tested was similar for all sterols. For a more physiological approach to in vivo conditions, incorporation of bile salts (10 mM), GL (60U/mL) and CE (0.075U/mL) to the INFOGEST method is proposed, although it increases the cost compared to the established method.

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 S phase, apoptosis induction and increase in caspases activation compared to 5-FU alone, while no significant effects were observed on ROS levels and ΔΨm. Our results suggest, for the first time, the ability of PS to sensitize colon cancer cells to 5-FU chemotherapy and warrant further in vivo studies to confirm our preclinical findings.

Plant sterol-enriched foods have proved to lower serum total and low-density lipoproteins-cholesterol concentration, so they have faced a great increase in the market over the past 20 years through accepted health claims. This chapter provides an overall and critical picture on the current knowledge and future perspectives regarding bioaccessibility and fermentation of sterols contained in plant sterol-enriched food products, particularly focused on the effect of food matrix composition, as well as different in vitro gastrointestinal digestion methodologies, and the role of microbiota in the sterol degradation. The final objective of this chapter is to evince the different aspects of the digestion and fermentation of plant sterol-enriched foods that require further research in order to improve the development of these functional foods and take advantage of their benefits.

Oxysterols are formed in foods during processing and storage, having an impact on the nutritional profile besides being associated to the development of diverse chronic and degenerative diseases. This review provides an overview on recent information and knowledge gaps on oxysterol formation and absorption from foods. Their contents should be monitored based on dietary habits, in order to have more realistic data available and to expand the currently limited daily intake data. Unraveling the mechanism of oxysterol absorption, together with a more thorough understanding of their metabolism, is needed; in this respect, bioavailability studies using in vitro and in vivo methodologies could contribute with new insights. Finally, the food-health axis can be boosted by suitable technological solutions and antioxidants to help preventing also in vivo oxidation.

The hypocholesterolemic effect of milk-based fruit beverages enriched with plantsterols (PS) has previously been demonstrated, obtaining a beneficial effect on the seruminflammatory status as well [1]. [...]

The consumption of milk-based fruit beverages enriched with plant sterols (PSs) has previously showed a cholesterol-lowering effect in postmenopausal women [1]. [...]

Unabsorbed cholesterol, along with that of bile secretions and flaked colon cells, can be metabolized by colonic microbiota. The generated metabolites have been proposed as promoters of colorectal cancer (CRC). In this study, the cytotoxicity (MTT assay) of the main commercially available cholesterol-derived metabolites (coprostanol, cholestanol, coprostanone, and cholestenone) on human colon cancer (Caco-2) and non-tumor (CCD-18Co) cells was evaluated at different physiologically relevant concentrations (9.4–300 µM) and incubation times (24–72 h). In general, the metabolites that most reduced cell viability were coprostanone (54–85% in Caco-2 and 20–81% in CCD- 18Co) and cholestenone (17–91% in Caco-2 and 14–81% in CCD-18Co). These two metabolites are the most hydrophobic, thus reflecting a possible relationship between hydrophobicity and cytotoxicity. Moreover, cholestenone (IC50 at 72 h: 5 ± 1 µg/mL) should be considered cytotoxic on CCD- 18Co cells (non-tumor cells) since it shows an IC50 close to the one considered toxic (

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 their BA ranged between 58 and 80% and 45–49%, respectively, without significant differences throughout storage. β-Sitosterol showed a lower mean oxidation percentage (0.028%) than cholesterol (1.24%), but the estimated POPs intake (0.5 mg/day) was two-fold higher than that of COPs (0.25 mg/day) from 250 g of beverage. These results show that the presence of milk fat and MFGM in the formulation of this beverage did not imply an increase in the contents of oxysterols and their BA. Thus, the beverage is suitable as a PS-enriched food matrix for the length of its shelf-life, and its consumption appears to be safe for consumers.

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 < 0.05) were detected. The cholesterol precursor lathosterol (11.2%), markers of the dietary PS intake (campesterol 43.1% and β-sitosterol 32.5%), and anti-inflammatory IL-10 cytokine (22.5%) increased significantly, with a concomitant significant reduction in pro-inflammatory IL-1β (6.7%). No variations in HDL-cholesterol, other sterols (desmosterol and stigmasterol) or cytokines (IL-6, IL-8, IL-12p70 and TNF-α) were detected. These results indicated that this kind of PS-enriched milk-based fruit beverage is suitable during the period of clinical intervention, and its consumption may be an adequate way to improve PS functionality since a significant reduction in cholesterol levels has been observed. Therefore, the intake of this beverage could contribute to reduce the risk of cardiovascular disease also obtaining a beneficial effect on the serum inflammatory status in postmenopausal women.

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, solid-phase microextraction or enzymes have also been applied. In addition, the determination of PS may also need a subsequent saponification, purification or enrichment step before their determination. Gas chromatography after derivatization has historically been the method of choice for analysis, although liquid chromatography simplifies the analysis. Furthermore, although various detectors have been employed in analysing PS, mass spectrometry can address many of the challenges observed in other methodologies. Therefore, it has been widely used in conjunction with other separation techniques due to its sensitivity and selectivity by the application of tandem mass spectrometry. This review provides an overall and critical picture of the current methodologies used for the analysis of PS in foods, particularly focused on the individual applications and limitations.

The impact of a plant sterol-enriched beverage on the sterol metabolism, organic acid production and microbiota composition was evaluated by means of a dynamic gastrointestinal and colonic fermentation model. After one week of fermentation, an absence of sterol metabolites was reported, in accordance with the lack of microbiota related to their metabolism. Although total organic acid content was lower in the ascending colon (AC) compared to the transversal (TC) and descending colon (DC) (28–57 mmol/L vs. 55-87 and 44–64 mmol/L, respectively), its increments, with respect to the initial value, were higher (2-fold vs. 1.6- and 1.5-fold). Increments of acetate, butyrate and propionate were observed in all colon vessels, whereas lactate production was only observed in the AC at the first hours of fermentation. Results showed a progressive increase in microbial species richness and evenness from the AC to the DC. Beverage fermentation also modulated certain members of the microbiota: Bifidobacterium (within 24 h) and Megasphaera genera were stimulated, and Mitsuokella was reduced in AC, while the growth of Klebsiella genus was promoted and Bacteroidetes phylum decreased in TC and DC. These results show that the beverage modulates the microbial community and activity, which could be physiologically relevant to the host.

Gangliosides (GGs) are important bioactive compounds that offer beneficial anti-infective, anti-inflammatory, and neuronal development effects. The present work reviews the techniques used to deter...

4th International Conference on Food Digestion -- MAR, 2015 -- Naples, ITALY, WOS: 000384789000006, Within the active field of in vitro digestion in food research, the COST Action INFOGEST aimed to harmonize in vitro protocols simulating human digestion on the basis of physiologically inferred conditions. A harmonized static in vitro digestion (IVD) method was recently published as a primary output from this network. To validate this protocol, inter-laboratory trials were conducted within the INFOGEST network. A first study was performed using skim milk powder (SMP) as a model food and served to compare the different in-house digestion protocols used among the INFOGEST members. In a second inter-laboratory study applying the harmonized protocol, the degree of consistency in protein hydrolysis was investigated. Analysis of the hydrolyzed proteins, after the gastric and intestinal phases, showed that caseins were mainly hydrolyzed during the gastric phase, whereas beta-lactoglobulin was, as previously shown, resistant to pepsin. Moreover, generation of free amino acids occurred mainly during the intestinal phase. The study also showed that a few critical steps were responsible for the remaining inter-laboratory variability. The largest deviations arose from the determination of pepsin activity. Therefore, this step was further clarified, harmonized, and implemented in a third inter-laboratory study. The present work gives an overview of all three inter-laboratory studies, showing that the IVD INFOGEST method has led to an increased consistency that enables a better comparability of in vitro digestion studies in the future. (C) 2015 The Authors. Published by Elsevier Ltd.

A beverage enriched with plant sterols (1 g/100 mL) and galactooligosaccharides (1.8 g/100 mL) was subjected to a dynamic gastrointestinal and colonic fermentation process to evaluate the effect on sterol metabolism, organic acid production, and microbiota composition. Production of sterol metabolites (coprostanol, methylcoprostanol, ethylcoprostenol, ethylcoprostanol, and sitostenone) was observed in the transverse colon (TC) and descending colon (DC) vessels in general, from 24 and 48 h, respectively. Microbial activity was assessed through the production of organic acids, mainly acetate in all colon vessels, lactate in the AC, and butyrate and propionate in the TC and DC. A higher diversity in the microbial community was found in the TC and DC, in accordance with a higher sterol metabolism and organic acid production. Although the prebiotic effect of galactooligosaccharides was not detected, changes in microbiota composition (an increase in the

International audience; Oat is rich in a wide range of phytochemicals with various physico-chemical, colloidal and interfacial properties. These characteristics are likely to influence human lipid metabolism and the subsequent effect on health following oat consumption. The aim of this work was to investigate the impact of oat materials varying in complexity on the lipolysis process. The composition, structure and digestibility of different lipid systems (emulsions, oil bodies and oil enriched in phytosterols) were determined. The surface activities of phytosterols were examined using the pendant drop technique. Differences in lipid digestibility of the oat oil emulsions and the oil bodies were clearly seen. Also, the digestion of sunflower oil was reduced proportionally to the concentration of phytosterols present. This may be due to their interfacial properties as demonstrated by the pendant drop experiments. This work highlights the importance of considering the overall structure of the system studied and not only its composition.

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 the development of functional PS-enriched milk-based fruit beverages considering: (i) PS ingredient selection; (ii) bioaccessibility: influence of matrix modification; (iii) bioavailability; and (iv) bioactivity. This integrated approach could serve as an example of the unequivocal demonstration of the efficacy of these functional beverages in promoting health benefits.

International audience; Serum concentrations of lathosterol, the plant sterols campesterol and sitosterol and the cholesterol metabolite 5alpha-cholestanol are widely used as surrogate markers of cholesterol synthesis and absorption, respectively. Increasing numbers of laboratories utilize a broad spectrum of well-established and recently developed methods for the determination of cholesterol and non-cholesterol sterols (NCS). In order to evaluate the quality of these measurements and to identify possible sources of analytical errors our group initiated the first international survey for cholesterol and NCS. The cholesterol and NCS survey was structured as a two-part survey which took place in the years 2013 and 2014. The first survey part was designed as descriptive, providing information about the variation of reported results from different laboratories. A set of two lyophilized pooled sera (A and B) was sent to twenty laboratories specialized in chromatographic lipid analysis. The different sterols were quantified either by gas chromatography-flame ionization detection, gas chromatography- or liquid chromatography-mass selective detection. The participants were requested to determine cholesterol and NCS concentrations in the provided samples as part of their normal laboratory routine. The second part was designed as interventional survey. Twenty-two laboratories agreed to participate and received again two different lyophilized pooled sera (C and D). In contrast to the first international survey, each participant received standard stock solutions with defined concentrations of cholesterol and NCS. The participants were requested to use diluted calibration solutions from the provided standard stock solutions for quantification of cholesterol and NCS. In both surveys, each laboratory used its own internal standard (5alpha-cholestane, epicoprostanol or deuterium labelled sterols). Main outcome of the survey was, that unacceptably high interlaboratory variations for cholesterol and NCS concentrations are reported, even when the individual laboratories used the same calibration material. We discuss different sources of errors and recommend all laboratories analysing cholesterol and NCS to participate in regular quality control programs.

This document is he Accepted Manuscript version of a Published Work that appeared in final form in Journal of Agricultural and Food Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.jafc.9b04796, A beverage enriched with plant sterols (1 g/100 mL) and galactooligosaccharides (1.8 g/100 mL) was subjected to a dynamic gastrointestinal and colonic fermentation process to evaluate the effect on sterol metabolism, organic acid production, and microbiota composition. Production of sterol metabolites (coprostanol, methylcoprostanol, ethylcoprostenol, ethylcoprostanol, and sitostenone) was observed in the transverse colon (TC) and descending colon (DC) vessels in general, from 24 and 48 h, respectively. Microbial activity was assessed through the production of organic acids, mainly acetate in all colon vessels, lactate in the AC, and butyrate and propionate in the TC and DC. A higher diversity in the microbial community was found in the TC and DC, in accordance with a higher sterol metabolism and organic acid production. Although the prebiotic effect of galactooligosaccharides was not detected, changes in microbiota composition (an increase in the Parabacteroides genus and the Synergistaceae and Lachnospiraceae families) indicated an enhancement of sterol metabolism.

Human milk (HM) is the exclusive food during the first 4–6 months of an infant's life. Breastfeeding has been related to significant health benefits for infants, and hence it is of interest to study the bioactive compounds present in HM, such as sterols (cholesterol being the most abundant). The aim of this study was to determine the contents of sterols (cholesterol, desmosterol, lathosterol, lanosterol, campesterol, stigmasterol and β-sitosterol) in 10 pools of colostrum, transitional milk, and 1, 3 and 6 month HM obtained from Spanish volunteers from two different geographical areas (coastal and central) and to estimate the intake and bioaccessibility (BA) of sterols in order to ascertain the fate of sterols after digestion. The results showed that the total sterol contents decreased to half the initial level during lactation (24–11 mg per 100 mL) and was significantly higher in samples from the coastal area. Total and animal sterol intakes were between 200 and 400 times higher than plant sterol intakes and were significantly higher in samples from the coastal area. However, no statistically significant differences were found in cholesterol and plant sterol intakes between areas. The BA of total sterols ranged from 45% to 69% and was higher in the first month, which coinciding with the highest fat content of milk. In conclusion, the sterol content varies depending on the lactation stage and the geographical area, and the BA of sterols can be positively affected by a higher lipid content. All these data may contribute to the development of infant formulas that are more similar to HM in terms of composition and behaviour after digestion, according to the lactation stage involved.

This study validates a gas chromatography (GC) method for determining the sterol profile of human milk (HM) and compares it with an enzymatic-spectrophotometric (E-S) method. Good linearity ( r > 0.97) and low limits of detection and quantification were obtained with the GC 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 GOS. The harmonized in vitro digestion model INFOGEST was applied to the MfB beverage, but the cholesterol content could not be quantified due to its contribution of bile salts. Hence, it was proposed: (i) a change in porcine bile salt concentration from 10 mM to 1.4 mM (in order to compare with micellar digestion); or (ii) a change of bile salt origin (bovine instead of porcine), maintaining physiological concentration (10 mM, INFOGEST condition). Both options allowed cholesterol quantification, with bioaccessibilities of 62% (reduction of bile salts) and 38% (replacement of the bile salt source), whereas plant sterol bioaccessibilities were 22% and 14%, respectively. Therefore, the change of bile salt origin maintaining INFOGEST concentration is proposed as a method to evaluate sterol (cholesterol and PS) bioaccessibility in these beverages, demonstrating the need for the selection of appropriate conditions of the INFOGEST harmonized method according to the food matrix and compounds to be determined.

The design of infant formulas (IFs) seeks to resemble human milk (HM) composition and functionality. The fat sources used usually comprise vegetable oil blends to mimic the fatty acid composition of HM and introduce changes in the animal/plant sterol ratio. In contrast, the use of milk fat globule membrane (MFGM)-rich ingredients could improve this aspect by increasing the ratio. The present study evaluates the bioaccessibility (BA) of sterols (cholesterol, desmosterol, brassicasterol, campesterol, stigmasterol, and β-sitosterol) in three IFs (with or without MFGM) using an in vitro digestion method simulating infant conditions. Analytical parameters confirmed the suitability of the method for all of these sterols. Results showed the presence of MFGM to increase cholesterol content (6-7 vs 2 mg/100 mL), this being the most bioaccessible sterol in the IFs. Although the BA of cholesterol was reduced in MFGM-enriched IF (65.6-80.4% vs 99.7%), the intake of bioaccessible cholesterol from these IFs was higher.

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 appearance of a sub-G1 cell population. Thus, the use of PS as functional ingredients in the development of PS-enriched foods could exert a potential preventive effect against human breast, colon and cervical cancer, although further in vivo studies are required to confirm our preclinical findings.

International audience; Increasing numbers of laboratories develop new methods based on gas-liquid and high-performance liquid chromatography to determine serum concentrations of oxygenated cholesterol metabolites such as 7alpha-, 24(S)-, and 27-hydroxycholesterol. We initiated a first international descriptive oxycholesterol (OCS) survey in 2013 and a second interventional survey 2014 in order to compare levels of OCS reported by different laboratories and to define possible sources of analytical errors. In 2013 a set of two lyophilized serum pools (A and B) was sent to nine laboratories in different countries for OCS measurement utilizing their own standard stock solutions. In 2014 eleven laboratories were requested to determine OCS concentrations in lyophilized pooled sera (C and D) utilizing the same provided standard stock solutions of OCS. The participating laboratories submitted results obtained after capillary gas-liquid chromatography-mass selective detection with either epicoprostanol or deuterium labelled sterols as internal standards and high-performance liquid chromatography with mass selective detection and deuterated OCS as internal standard. Each participant received a clear overview of the results in form of Youden-Plots and basic statistical evaluation in its used unit. The coefficients of variation of the concentrations obtained by all laboratories using their individual methods were 58.5-73.3% (survey 1), 56.8-60.3% (survey 2); 36.2-35.8% (survey 1), 56.6-59.8, (survey 2); 61.1-197.7% (survey 1), 47.2-74.2% (survey 2) for 24(S)-, 27-, and 7alpha-hydroxycholesterol, respectively. We are surprised by the very great differences between the laboratories, even under conditions when the same standards were used. The values of OCS's must be evaluated in relation to the analytical technique used, the efficiency of the ample separation and the nature of the internal standard used. Quantification of the calibration solution and inappropriate internal standards could be identified as major causes for the high variance in the reported results from the different laboratories. A harmonisation of analytical standard methods is highly needed.

This study for the first time compares the in vivo (response in serum from individuals of an interventional study) and in vitro effects (bioaccessibility determined from simulated gastrointestinal digestion) of two plant sterol (PS)-enriched milk-based fruit beverages (with and without tangerine fruit juice rich in β-cryptoxanthin (β-Cx)) in order to assess their suitability as PS-enriched food matrixes, and the possible interactions between the two bioactive compounds (β-Cx and PS) that might affect their absorption. The randomized, double-blind, crossover study (performed in 36 postmenopausal women) showed the β-sitosterol and campesterol serum concentrations after four weeks of consumption of the beverages (1.5 g PS/day) to increase significantly (ranging 1.6–2.4 µg/mL and 0.3–0.4 µg/mL, respectively), thereby confirming suitability as matrices for PS-enrichment. The results therefore indicated similar behavior of PS supplied by these beverages and evaluated through in vitro or in vivo assays, independently of whether the samples contained β-Cx or not.

Sterols are components present in the fat fraction of infant formulas (IFs). Their characterization is therefore of interest, though there are no official reference methods for their analysis in these matrices.Aim: To validate a gas chromatographic method with flame ionization detection for the determination of animal (cholesterol and desmosterol) and plant sterols (brassicasterol, campesterol, stigmasterol, β-sitosterol and sitostanol) found in IFs.All correlation coefficients obtained for the calibration curves of sterols studied were >0.99. Limits of detection (

In this study the quality characteristics and content of healthy minor components of four crude canola oils as an effect of different oil extraction method (solvent extraction, hot pressing, and cold pressing) were studied. Cold-pressed canola oils had lower concentrations of FFA, PV, p-AV and chlorophylls than solvent-extracted, and hot-pressed canola oils. Oils obtained via the different extraction methods had different fatty acid profiles as well as dissimilar amounts of tocopherols, phytosterols, and polyphenols. The amount of total tocopherols in solvent-extracted canola oil was 493 mg/kg compared to 388 mg/kg for hot-pressed canola oil. The tocopherol content for two other cold-pressed and one other RBD canola oil was 366, 354, and 327 mg/kg, respectively. Solvent-extracted canola oil exhibited the highest free phytosterol content (178 mg/100 g), while RBD canola oil only had 129 mg/100 g of free phytosterols. While cold-pressed canola oil had the lowest amount of polyphenols, traditional refining resulted in almost complete removal of polyphenols from canola oil.