Guadalupe Garcia-Llatas

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 of oxidation in the ingredients tested, it can be concluded that the PS remain stable in these ingredients. Significant correlations (p < 0.01) were found between total oxysitosterols versus β-sitosterol contents (R(2) = 86.5%) and between total POPs and total PS (R(2) = 81.6%).

Sterol Oxidation in Ready-to-Eat Infant Foods During Storage

The effect of storage on sterol oxidation of ready-to-eat infant foods was evaluated. Two different liquid infant foods (honey or fruits flavors), prepared with milk and cereals, were stored for 0, 2, 4, 7 and 9 months at 25 degrees C. Sterol oxidation products (SOP) were isolated by cold saponification, purified by silica solid-phase extraction, and analyzed by gas chromatography (GC) and GC-mass spectrometry. beta-Sitosterol was the most representative sterol, followed by cholesterol and campesterol. No significant differences in the total and single SOP content (0.8-1 mg/kg of product) were observed with respect to storage time and type of sample; the main SOP found was 7-ketositosterol (<0.2 mg/kg of product). The extent of stigmasterol oxidation (2.9%) was higher than that of cholesterol (1.9%) and beta-sitosterol (1.4%). The type and quality of raw materials, as well as the processing conditions, seem to greatly influence SOP formation and accumulation in infant foods.

7-Ketocholesterol as marker of cholesterol oxidation in model and food systems: when and how.

Cholesterol can undergo oxidation through enzymatic or chemical mechanisms, generating a wide range of oxidation products (COPs) with adverse biological effects. COPs are characterized by different functional groups and are produced in different ratios or amounts, depending on the treatment and storage conditions. To follow the cholesterol oxidation process, 7-ketocholesterol (7-KC) has been often used as an oxidation marker in both model and food systems, since it is easily formed and is one of the most representative ring COPs. However, 7-KC does not always rise with increasing time/temperature conditions, especially in complex systems and high-protein or extensively processed foods. The following review provides a critical picture of the utilization of 7-KC as a cholesterol oxidation marker in model and food systems, focusing on the possible causes and effects of the different behaviours and trends, as well as on the advantages and disadvantages of using 7-KC when the extent of cholesterol oxidation is to be assessed.

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

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. Serum β-Cx and PS were determined by UPLC and CG-FID respectively. Outcome variables included markers of bone turnover and cardiovascular risk. Biological effect was assessed by paired t test and generalized estimating equations analysis that included the previous treatment, the order of intervention and the interactions. The intake of beverages containing β-Cx and PS brought about a significant increase in serum levels of β-Cx, β-sitosterol and campesterol. Intervention caused changes in almost all the markers while the order, previous treatment and the interaction did not reach statistical significance. Only the intake of the beverage containing β-Cx plus PS brought about significant decreases in total cholesterol, c-HDL, c-LDL and bone turnover markers. CONCLUSIONS β-Cx improves the cholesterol-lowering effect of PS when supplied simultaneously and this combination may also be beneficial in reducing risk of osteoporosis. CLINICAL TRIAL REGISTRY ClinicalTrials.gov number NCT01074723.

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 color over time and temperature, probably due to Maillard reaction compound formation. The increase in total antioxidant capacity might have helped PS maintenance throughout storage, these beverages being a good PS source even after 6 months of storage.

Study of thermal resistance and in vitro bioaccessibility of patulin from artificially contaminated apple products.

Apple juices and purees represent categories widely consumed by whole population and above all children. Patulin (PAT) is a mycotoxin known for its acute and chronic effects in animals. Several studies indicate there is a risk associated to the PAT intake, through the consumption of purees and apple juices. In this study, apple juice and puree were prepared and artificially contaminated with PAT at 50 μg/kg and submitted to a thermal treatment simulating pasteurization to evaluate PATs reduction. In a second phase of the work, apple products samples (n=7) included juices, nectars and purees belonging to different commercial brands were collected, artificially contaminated with PAT at 50 μg/L (limit established for PAT in juices) and 25 μg/kg (limit established for PAT in purees), digested with an in vitro gastrointestinal protocol and bioaccessibility values (%) were calculated. After thermal treatment, the PATs loss evidenced in purees and juices was of 1.41 ± 0.52% and 62.62 ± 2.53% respectively. Related to the bioaccessibility data, two juices with pulp showed values of 70.89 ± 4.93 and 67.30 ± 10.76%; two purees showed levels of 58.15 ± 5.50 and 55.69 ± 4.73%, whereas nectar and two clarified juice showed percentages of 38.88 ± 2.42, 28.59 ± 0.46 and 25.28 ± 0.61%, respectively.

Dietary phytochemicals in the protection against oxysterol-induced damage

The intake of fruits and vegetables is associated with reduced incidence of many chronic diseases. These foods contain phytochemicals that often possess antioxidant and free radical scavenging capacity and show anti-inflammatory action, which are also the basis of other bioactivities and health benefits, such as anticancer, anti-aging, and protective action for cardiovascular diseases, diabetes mellitus, obesity and neurodegenerative disorders. Many factors can be included in the etiopathogenesis of all of these multifactorial diseases that involve oxidative stress, inflammation and/or cell death processes, oxysterols, i.e. cholesterol oxidation products (COPs) as well as phytosterol oxidation products (POPs), among others. These oxidized lipids result from either spontaneous and/or enzymatic oxidation of cholesterol/phytosterols on the steroid nucleus or on the side chain and their critical roles in the pathophysiology of the abovementioned diseases has become increasingly evident. In this context, many studies investigated the potential of dietary phytochemicals (polyphenols, carotenoids and vitamins C and E, among others) to protect against oxysterol toxicity in various cell models mimicking pathophysiological conditions. This review, summarizing the mechanisms involved in the chemopreventive effect of phytochemicals against the injury by oxysterols may constitute a step forward to consider the importance of preventive strategies on a nutritional point of view to decrease the burden of many age-related chronic diseases.

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 proposed in beverages of this kind to ensure optimum bioavailability of PS. Our results suggest that the BA of PS is influenced by the type and quantity of fat and the emulsifier type involved.

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≫FJPS>MFJPS. In general, the beverages showed low PS oxidation levels (<0.17%). Predictive models of POP content versus storage time were established. These models explain total POP content by over 75% and individual POP content by over 50%. We propose 7-ketositosterol and 7-ketocampesterol as PS oxidation markers during storage of beverages of this kind.

Static Digestion Models: General Introduction

Several in vitro methods have been developed to simulate the physiological conditions of the human gastrointestinal digestion, the simplest being the static methods. The following chapter clarifies the concepts of bioaccessibility and dialyzability, and describes the conditions (pH, enzymes, agitation, etc.) to be applied in oral, gastric and intestinal phases when assessing a food component (nutrient, bioactive or toxin) or a food product, in a single or multi-phase model. The advantages and disadvantages of the static models vs. dynamic and in vivo models are discussed, and a review of specific conditions applied on nutrients (minerals, vitamins, proteins, fatty acids, etc.) and bioactive compounds (carotenoids, plant sterols, etc.) from recent studies is provided. Currently, it must be considered that, although the static digestion conditions must be adapted according to the component or food sample to be studied, a harmonization and standardization of the models are needed in order to establish suitable correlations among in vitro and in vivo assays, as it has been defined for some food components (carotenoids, proteins and minerals).