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To exploit the full potential of pomegranate marcs, this study explores different conditions of ‘green’ extraction of ellagitannins and other bio-active phenolic compounds from the ‘external by-product’ of pomegranate industrial processing. Significant recoveries of ellagitannins, free ellagic acid and radical scavenging compounds were obtained by coupling both the traditional (MH) or the hydro-diffusion gravity (MHG) microwave assisted drying to a direct ultrasound assisted extraction (UAE) using a food-grade solvent (EtOH/H2O). The amounts of the ellagitannins after microwave drying were always significantly higher than with the traditional oven drying. Moreover, microwaves reduced the drying times from several hours to few minutes reaching comparable residual moisture content. The microwave hydro-diffusion gravity system allowed the recovery of bio-active compounds in the condensed water too. The comparison of UAE with traditional maceration and decoction highlighted the higher efficiency of UAE in the quicker extraction of bio-active compounds, except for the total ellagitannins which were better recovered after the longer maceration.

FINNOVER is an EU Interreg-Alcotra project that aims to bring new perspectives to floriculture enterprises by recovering useful bioproducts from the waste produced during processing of several aromatic species. In this study, a new operation strategy to recover lavender (Lavandula angustifolia Mill.) solid by-products remaining after the extraction of the essential oil was developed. Pulsed ultrasound-assisted extraction was employed as a sustainable and eco-compatible technology to extract, in a very short time (10 min), this agricultural waste using a food-grade solvent (a mixture of ethanol/water). All the extracts obtained from both flower and leaf waste and flower-only residues, exhibit a promising total phenolic content (38–40 mg gallic acid/g of dry waste), radical scavenging activity (107–110 mg Trolox/g of dry waste) and total flavonoid content (0.11–0.13 mg quercetin/g of dry waste). Moreover, the chromatographic analysis of these extracts has shown that this overlooked agriculture waste can represent a valuable source of multifunctional compounds. Particularly, they exhibit a content of polyphenols and flavonoids up to 200 times higher than the corresponding leachate, and they are a valuable source of gentisic acid (1.4–13 mg/g dry waste) representing a new low-cost ingredient usable in different fields (i.e., cosmetic).

International audience; Oryza sativa L. 'Violet Nori' is an Italian cultivar of spontaneous growing aromatic purple rice which is particularly rich in polyphenolic compounds, especially anthocyanins, conferring it an excellent antioxidant capacity. The present study aimed at increasing the extraction yields of its antioxidant compounds with green strategies and it is divided into two steps. The first step concerned a solubility study of the targeted polyphenols in different ethanol:water mixtures by means of a theoretical prediction method, using the simulation program COSMO-RS, and the subsequently confirmation of the computational results by practical experiments. Once the best extraction mixture was identified, the second step of the work was performed, with the purpose of intensifying the extraction yield. Therefore, various innovative green extraction techniques, including ultrasound, using both the probe system and the ultrasonic bath, bead milling, microwave and accelerated solvent extractions were tested and compared to conventional maceration.Results, expressed in terms of total phenolic and total monomeric anthocyanin contents, showed that the best extracting solvent for 'Violet Nori' rice was the mixture ethanol:water (60:40 v/v), being COSMO-RS computational predictions in good correlation with the experimental results. Moreover, the most efficient techniques to extract the antioxidant compounds resulted to be both ultrasound-assisted extraction probe and bead milling, that in only 5 min got the same extractive efficiency obtained after 3 h of conventional maceration.

Pomegranate fruit is recognized as an important source of health compounds and its juicy arils represent the most commercially exploitable part of the fruit. Nevertheless, by-products coming from pomegranate juice processing represent both a challenging disposal problem and a promising source of health ingredients. In particular, the marcs obtained after juice squeezing are usually discarded or used for oil production in virtue of their lipophilic fraction, although the residues of pericarp and arils are proven to still be a good source of hydrophilic compounds such as polyphenols. Pulsed ultrasound-assisted extraction, using just water as solvent, and spray-drying microdispersion, using low methoxyl pectin as polymeric matrix, have been employed, respectively, to extract and formulate the water-soluble bioactive molecules from these by-products. From 100 g of pomegranate fresh marcs, almost the same quantity of phenolic compounds found in 100 mL of the corresponding juice can be extracted with similar antioxidant activity, but with higher content in vitamin C and practically without total soluble solids. The extracts have been sprayed obtaining powders with an encapsulation efficiency of about 50%. The extracts, both before and after the microparticles production, have been characterized and their capacity in inhibition of platelets aggregation induced by thrombin has been tested ex vivo on human platelets (% inhibition: about 60% and 30% before and after microencapsulation, respectively). Finally, fresh-cut apple wedges were enriched by vacuum impregnation with the formulated extracts, tentatively used as potential novel ingredients, obtaining “polyphenol-enriched” apples.

The immune system and the central nervous system message each other to preserving central homeostasis. Both systems undergo changes during aging that determine central age-related defects. Ellagic acid (EA) is a natural product which is beneficial in both peripheral and central diseases, including aging. We analyzed the impact of the oral administration of a new oral ellagic acid micro-dispersion (EAm), that largely increased the EA solubility, in young and old mice. Oral EAm did not modify animal weight and behavioral skills in young and old mice, but significantly recovered changes in &ldquo, ex-vivo, in vitro&rdquo, parameters in old animals. Cortical noradrenaline exocytosis decreased in aged mice. EAm administration did not modify noradrenaline overflow in young animals, but recovered it in old mice. Furthermore, GFAP staining was increased in the cortex of aged mice, while IBA-1 and CD45 immunopositivities were unchanged when compared to young ones. EAm treatment significantly reduced CD45 signal in both young and old cortical lysates, it diminished GFAP immunopositivity in young mice, but failed to affect IBA-1 expression in both young and old animals. Finally, EAm treatment significantly reduced IL1beta expression in old mice. These results suggest that EAm is beneficial to aging and represents a nutraceutical ingredient for elders.

Ellagic acid (EA) is a bioactive natural polyphenol with antioxidant and several other beneficial properties, but unfortunately it cannot be exploited for in vivo therapeutic applications because of its poor water solubility and accordingly low bioavailability. Aiming to increase EA solubility, an EA solid microdispersion was realized by employing only water and low methoxylated pectin as a food compatible excipient by applying spray drying technology. It showed a 22% (w/w) Drug Loading (DL), a 30 times improved water solubility and remarkable radical scavenging activity. Subsequently, adopting non-PAMAM hydrophilic and amphiphilic dendrimers as nanocontainers, two EA nanodispersions were achieved (60–70 nm) with 46 and 53% (w/w) DL, water solubility 300 to 1000 times higher than that of free EA and good antioxidant capability. In both cases, toxic agents such as castor oil, Tween 80 or the extensively used but not so safe PEG400 were avoided and no previously used carriers, including β-cyclodextrins, were adopted. The prepared micro- and nanodispersions showed water solubility never obtained before and represent non-toxic EA reservoirs suitable for food and biomedical applications.

In this research, for the first time, an alternative method to produce Ribes nigrum bud derivatives is presented. Pulsed ultrasound-assisted extraction (PUAE), using a food-grade solvent according to green chemistry principles, has been employed and compared to the conventional extraction method. Traditionally, bud derivatives, a category of botanicals marketed as plant food supplements in the European Community, are produced by macerating meristematic tissues of trees and plants mainly spontaneously collected. Buds are a challenging raw material for the UAE, since meristematic tissues are much softer and fragile than their corresponding adult phenological stage. It is therefore important to assess whether the polyphenolic fraction, very susceptible to degradation, is conserved after UAE. Untargeted polyphenolic fingerprints (UV-Visible and fluorescence) coupled with chemometrics are employed to quickly screen the best extraction conditions, evaluated by the design of experiment (DoE) method. The polyphenolic fraction of the optimized PUAE extract was quantified by targeted HPLC fingerprint and its antiradical activity was determined. PUAE on a lab pilot reactor was proven to be the most practical approach for a rapid (20 min vs. 21 days maceration) and efficient extraction of bioactive polyphenolics from Ribes nigrum buds, encouraging the scale up to an industrial plan.

The study is focused on the effect of different cooking conditions on the antioxidant content, particularly anthocyanins, of Oryza sativa L. ‘Violet Nori’, a new black rice cultivar. 10 different cooking tests were performed. The selected cooking conditions allowed to evaluate the effect of boiling, roasting, oven cooking, risotto cooking, and oriental cooking. The total anthocyanins amounts were evaluated by both a spectrophotometric pH differential method and High-Performance Liquid Chromatography (HPLC), together with the Total Phenolic Content (TPC) and the Radical Scavenging Activity (RSA). The obtained results showed that boiling with a low water amount (100 g rice/650 mL water) and oriental cooking, which allows a reduction of boiling times thanks to the lid of the pot, allow effectively saving at least part of their anthocyanins content. Similar results were obtained by boiling if rice was introduced in cold or boiling water, but on the contrary a ratio of 100 g rice/1000 mL water greatly enhanced the loss of antioxidant compounds. Risotto and oven cooking allowed obtaining results that were roughly intermediate between the ‘best’ and the ‘worst’ results. A high correlation existed between the amounts of total anthocyanins and both TPCs and RSAs. The ‘best’ results show that although cooking necessarily decreases the rice content of valuable antioxidants, a careful choice of the operative conditions allows effectively preserving amounts of total anthocyanins higher than 100 mg/100 g rice portion, which are close to or even higher than in other well-known sources of dietetic anthocyanins.

It is well-known that the health properties attributed to several fruits, herbs, seeds and their processed foods/beverages are due to an important group of natural polyphenols classified as hydrolysable tannins (HT) named ellagitannins (ETs), that encompass both one or more gallic acid (GA) units and one or more hexahydroxydiphenoic acid (HHDP) units, ester-connected with a sugar residue. In vivo, ETs are rather not absorbed and in gastrointestinal tract (GIT), they are hydrolysed providing mainly ellagic acid (EA). Due to its trivial water-solubility, first pass effect, metabolism in GIT, or irreversible binding to cellular DNA and proteins, EA has a very low bioavailability. Some authors are studying methods to increase EA water-solubility and thus to improve its bioavailability. At the same, EA metabolism to urolithins (UROs), whose concentration and activity is inter-individual and intra-individual dependent, is still under study and not completely elucidate. Numerous in vitro and in vivo studies have been carried out to define the molecular and cellular events underlying the beneficial effects that this compound and its metabolites exert in pathological conditions. The anti-inflammatory and the antioxidant properties of EA attracted the interest of researchers for its potential health benefits in humans, including anti-cancer, anti-diabetes activities and cardio-protection. Nevertheless, lately the attention paid to EA is focusing on its potential protective action towards several neurodegenerative disorders. Thus, EA is investigated as a potential “lead compound” endowed with multi-target pharmacological properties on CNS. Since the identification of the pharmacophore(s) responsible for both health benefits and collateral effects of this compound is crucial in drug discovery, this review aims to provide an all-round updated analysis of the literature concerning EA involvement in several CNS disorders, hoping that such information will be useful to researchers involved in multi-target drug design for CNS.

This research takes place in the context of an Alcotra Italy-France trans-frontier project called FINNOVER, which includes among its objectives the “green” innovation of agro-industrial chains. Bud-derivatives are a category of natural products produced macerating meristematic tissues of trees and plants. They are quite expensive compared to other botanicals, since the collection period of their raw materials is extremely limited over the time. Pulsed Ultrasound-Assisted Extraction has been employed to extract further valuable material from the buds by-products remaining after the production of Castanea sativa Glyceric Macerates. UV–Visible spectra coupled with chemometrics were employed, as untargeted phytochemical fingerprints, to quickly screen the best experimental conditions of extraction: a duty cycle of 80%, an extraction time of 15 min and a solvent/ratio of 1/10. Targeted phytochemical fingerprints by HPLC have been used to identify and quantify the main bioactive compounds of the most promising marcs extract comparing it with the corresponding commercial Castanea sativa Glyceric Macerate. An innovative extraction and re-use strategy to obtain value-added products from botanicals by-products was developed in alternative to incineration or composting. It was applied to Castanea sativa buds production as case study, but it could be analogously applied for other herbal preparations.

The aim of this paper was to study the fatty acid composition of total and "free" lipids in powdered infant milks and to evaluate the fatty acid oxidation after package opening. In its first step the application of Principal Component Analysis and Design of Experiments has allowed to take a comprehensive picture of powdered infant formulas on sale in Italy on the basis of their labelled composition and to select a limited number of representative samples. Both total and "non encapsulated" or "free" lipids were considered and analysed and no significant differences were found between their fatty acids contents. The possible modification of the fatty acid composition in the "free" lipids caused by oxidation was also studied, simulating the domestic habits after opening. No significant changes were observed up to 30 days even when formulas had an high content of unsaturated fatty acids and valuable Long Chain Polyunsaturated fatty acids.

‘Violet Nori’ is a new spontaneous growing violet rice variety showing a peculiar violet color in its fresh leaves as well. In this paper, the antioxidant properties and the content of total phenols, radical scavengers, and anthocyanins in ‘Violet Nori’ caryopses, flour, and leaves are explored and compared. Ultrasonic-Assisted Extraction (UAE) is employed for the extraction of phenolic compounds, improving the extraction conditions by Design of Experiments (DoE). The obtained results show that the Radical Scavenging Activities (RSAs), the Total Phenolic Contents (TPCs), and the anthocyanin amounts (1000–1500 μg/g, expressed as cyanidin-3-glucoside) of ‘Violet Nori’ caryopses are higher than those in the other analyzed colored rice samples (300–900 μg/g as cyanidin-3-glucoside), with the exception of the cultivars ‘Artemide’ and ‘Nerone’, which show comparable values of RSAs and TPCs. The study of ‘Violet Nori’ leaves at different plant maturation stages shows that their anthocyanin content is 2–3 times higher than in the caryopses and in the flour, reaching the highest levels at about 60 days from seeding. Thus, the estimated extraction yield of 4 kg anthocyanins/t makes fresh leaves very interesting for the extraction of anthocyanins on an industrial scale, whereas violet caryopses are a very interesting dietetic source of valuable anthocyanins and other antioxidant compounds.

The aim of this study was to investigate the potential of retrieving polyphenolic antioxidants directly from wet pomegranate marcs: the fresh by-products obtained after pomegranate juice processing. These by-products mainly consist of internal membranes (endocarp) and aril residues. Even if they are still edible, they are usually discharged during juice production and, thus, they represent a great challenge in an eco-sustainable industrial context. Green technologies, such as ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE), have been employed to convert these organic residues into recycled products with high added value. UAE and MAE were used both in parallel and in series in order to make a comparison and to ensure exhaustive extractions, respectively. Water, as an environmentally friendly extraction solvent, has been employed. The results were compared with those ones coming from a conventional extraction. The most promising extract, in terms of total polyphenol yield and radical scavenging activity, has been tested both as a potential natural additive and as a functional ingredient after its incorporation in a real food model and in a real cosmetic matrix, respectively. This study represents a proposal to the agro-alimentary sector given the general need of environmental “responsible care”.

In this study vacuum impregnation (VI) was employed for the iron enrichment of olive fruits, which are very interesting as food vehicle for VI mineral supplementation for the porosity of their pulp. NaFeEDTA was chosen for olives fortification since it prevents iron from binding with compounds that could hinder it from being efficiently absorbed and since it causes few organoleptic problems. In order to improve the efficiency of the VI process, several parameters of the whole process were studied by design of experiment techniques. First of all D-optimal design was employed for a preliminary screening of the most significant process variables and showed that the concentration of VI solution was by far the most significant process variable, though its time in contact with olives was also significant. A factorial design was then applied to the remaining variables and it showed that the speed of the addition of VI solution was also significant. Finally, the application of a face centered composite design to the three selected variables allowed to detect processing conditions leading to final iron contents of 1.5–3 mg/g, corresponding to an introduction of 10–15 mg Fe with four or five fortified olive fruits. No effect on olive taste was observed at these concentrations. The results showed that olive fruits were the most interesting vehicles for the supplementation of both iron and other minerals.

Starting from a registered patent, the aim of the present study was to improve the osmotic dehydration (OD) process of Protected Designation of Origin Genovese basil leaves by composite experimental design. The contemporary optimization of both process variables and mixture composition allowed enhancing the quality of the dried product. A tentative substitution in the OD brine of the polyalcohol sorbitol with the novel food trehalose was also performed. Sensorial techniques and headspace analysis were applied to monitor changes in appearance, texture, flavor and color. The best appearance and aroma of dried leaves were obtained at 67.5C with a dipping time of 55 s in a brine containing 65% water, 20% sorbitol and 15% NaCl. Using trehalose did not lead to a significant improvement of the quality of the final product. This experimental approach could be successfully used, also on industrial scale, for the optimization of similar OD processes. PRACTICAL APPLICATION The paper shows the optimization of an osmotic dehydration (OD) process by composite design in order to simultaneously optimize both the process variables and the brine composition. This experimental approach can be successfully used, also on industrial scale, for the optimization of similar OD processes, with the aim to enhance the quality of the dried food and to limit their changes in appearance, texture, flavor and color.

An authentication study of the Italian PDO (protected designation of origin) extra virgin olive oil Chianti Classico was performed; UV–visible (UV–vis), Near-Infrared (NIR) and Mid-Infrared (MIR) spectroscopies were applied to a set of samples representative of the whole Chianti Classico production area. The non-selective signals (fingerprints) provided by the three spectroscopic techniques were utilised both individually and jointly, after fusion of the respective profile vectors, in order to build a model for the Chianti Classico PDO olive oil. Moreover, these results were compared with those obtained by the gas chromatographic determination of the fatty acids composition. In order to characterise the olive oils produced in the Chianti Classico PDO area, UNEQ (unequal class models) and SIMCA (soft independent modelling of class analogy) were employed both on the MIR, NIR and UV–vis spectra, individually and jointly, and on the fatty acid composition. Finally, PLS (partial least square) regression was applied on the UV–vis, NIR and MIR spectra, in order to predict the content of oleic and linoleic acids in the extra virgin olive oils. UNEQ, SIMCA and PLS were performed after selection of the relevant predictors, in order to increase the efficiency of both classification and regression models. The non-selective information obtained from UV–vis, NIR and MIR spectroscopy allowed to build reliable models for checking the authenticity of the Italian PDO extra virgin olive oil Chianti Classico .

Modern membrane technologies are useful for enhancing the concentration of phenolic antioxidants in olive mill waste water (OMWW) to produce concentrates with valuable applications in functional foods. Three types of OMWW concentrates, each with different levels of solute concentration and purity, were obtained from a single OMWW batch and dissolved in two extra virgin olive oils to achieve saturated solutions. Three addition levels were considered. Accelerated aging testing of the oils was performed at 60 °C and the samples were analyzed after two, four, and 6 weeks of aging. D-optimal design was used to select the 26 experiments that allowed the evaluation of the influence of the different variables on oil stability. The addition of OMWW concentrates resulted in a significant increase in the Radical Scavenging Activity (RSA) of the olive oils. Under these mild experimental conditions, the moderate formation of fatty acid hydroperoxides was probably masked by interfering compounds.

Headspace sorptive extraction and GC/MS, coupled with chemometric tools, were used to predict the amounts of pine nuts and Pecorino in Pesto Genovese, a typical Italian basil-based pasta sauce. Two groups of samples were prepared at different times and with ingredients from different batches for building the predicting models and testing their performances. Principal component analysis and partial least-squares regression (PLS) were applied to the chromatographic data. The 24 most-predictive variables were selected, and the application of PLS to the training set samples led to two models that explained approximately 70 of the variance in cross-validation, with prediction errors of 0.1 g for Pecorino and 0.6 g for pine nuts, thus confirming the reliability of the analytical method and the predicting ability of the models. The results obtained for the test set samples were not completely satisfactory, with a prediction error and a bias of 5.0 and 4.1 g, respectively, for Pecorino and corresponding values of 4.1 and 2.0 g for pine nuts. This preliminary study shows that the analytical methods used can allow construction of models with high predictive ability only if the great variability of the headspace composition of the ingredients and the effect of Twister are considered.

This paper describes the development and application of a new analytical method for the analysis of the volatile fractions of pesto , a typical Italian basil-based pasta sauce, and its ingredients. The extraction of the volatile fraction was realized by headspace sorptive extraction (HSSE) and the extracted volatiles were then thermodesorbed, cryoconcentrated in a cooled injector and analysed by gas chromatography-mass spectrometry. This study shows that despite the great variability detected in the composition of the volatile fractions of several samples of “Pesto Genovese”, the ingredients traditionally used in pesto preparation (basil, pine nuts, walnuts, cheese, extra-virgin olive oil and garlic) give a significant and distinctive contribution to the pesto volatile fraction. Thus, the HSSE analysis of “Pesto Genovese” could be very useful for a careful control of the composition of this valuable niche product.

Cholesterol oxidation in commercial meat-based homogenized and freeze-dried baby foods was examined. The 7 major products of this reaction were determined by gas chromatography coupled with mass spectrometry (GC-MS). As far as single cholesterol oxidation products (COP) are concerned, 7-ketocholesterol was the major product of direct cholesterol oxidation in the 2 groups of analyzed samples, and this study confirmed that it is a useful marker of the whole cholesterol oxidation process. Nevertheless, the amounts of cholesterol-5β,6β-epoxide were often similar to and sometimes higher than the amounts of 7-ketocholesterol, thus showing a strong development of both direct and indirect cholesterol oxidation pathways. Total COP content was significantly higher in freeze-dried than in homogenized products. Moreover, in freeze-dried samples, the COP content per serving was quite variable and, in 2 samples, it was close to or even higher than 500 μg. The greater development of cholesterol oxidation in the freeze-dried samples was confirmed by their highest total COP/cholesterol percent ratios. A constant correlation between the fatty acid composition and the development of cholesterol oxidation was not found, although a positive correlation between unsaturated fatty acid content and total COP content occasionally exists in samples of the same brand.

The variations in the carbohydrate and essential amino acid contents in grains from durum and common wheat, barley, rye and triticale during development were analyzed. The quantities of simple sugars and fructans reached their highest levels approximately ten days after anthesis, remained high for few days and then they decreased again. Among amino acids, the most significant variations were recorded for leucine and cysteine, which increased, and for histidine and lysine, which decreased. From the nutritional point of view meals from immature grains have a high content of lysine, fructans and simple sugars. They could thus be particularly significant for the production of functional foods.

Both consumers and the food industry are increasingly interested in aromatic herbs, not only as flavoring agents, but also for their content in biologically active compounds. Sweet basil (Ocimum basilicum L.) is a very important culinary herb and a functional ingredient marketed fresh, dried, or frozen. At present, drying is by far the most widely used basil preservation method, thus ensuring microbiological safety and extending basil shelf-life. Nevertheless, during drying, a series of physical and chemical changes that may have an adverse effect on basil quality may take place. Such alterations include changes in its flavor and appearance, mainly caused by the loss of volatile components and the formation of new compounds. Both these aspects are important quality factors with a deep impact on the consumers’ acceptance of the product. In this chapter the most commonly used drying methods are described and their impact on the sensorial and nutritional properties of this raw material are analyzed.

Cholesterol oxidation in tuna canned in brine was studied. Gas chromatography-mass spectrometry was used for the detection of the seven major cholesterol oxidation products originating from both direct and indirect oxidation. The total amount of cholesterol oxidation products in the analyzed samples varied considerably, ranging between 40 and 350 μg/g lipids, with the exception of an anomalous sample, that reached a 1600 μg/g level. The lipid content ranged between 0.5 and 1 g/100 g wet product. As most samples did not exceed 100 μg/g lipids, it is possible to assume that the total content of cholesterol oxidation products can be kept below this value if good manufacturing conditions are used, together with a careful choice of the best tuna cuts. The application of principal component analysis to the detected variables confirmed that 7-ketocholesterol is a useful index of the whole oxidation process.

A fast and reliable method for the determination of free amino acids in infant formulas was developed. The method emphasizes the determination of taurine, methionine, arginine and tyrosine, as they are often added to these formulas in order to adapt them to the nutritional requirements of infants. The method involves the precipitation of the protein fraction, its separation by centrifugation, and the HPLC analysis of the o -phthaldialdehyde (OPA) derivatives of the free amino acids. The method was applied to several commercial formulas, as their actual content of free amino acids may be reduced by the development of Maillard reaction during processing and storage. The detected amounts of free taurine and methionine were often lower than those listed by the manufacturers. In some samples significant amounts of free arginine, tyrosine and methionine were detected, although they had not been listed among the added ingredients.

The development of the Maillard reaction in pasteurized, UHT and in-bottle sterilized dietetic milks was studied. In these products damage caused by heat treatments could increase as a result either of the addition of various ingredients or of manufacturing processes that alter their content of reducing carbohydrates. Protein damage was evaluated by measuring furosine by reversed-phase ion-paired HPLC. The levels of furosine detected made it possible to assess the amounts of biologically unavailable lysine. In all milks analysed blocked lysine values were

Polar compounds of virgin olive oils were analyzed. They influence oil flavor and aroma and improve the shelf-life of the oil. The orthodiphenolic fraction is particularly significant for oil stability because of its antioxidative activity. A relationship between the composition of the whole fraction of polar compounds and the state of health of the olives was established. For this purpose, oil samples were obtained from olives that had reached different degrees of ripeness and that had been affected by Dacus oleae infestation differently. The polar compounds were then analyzed by high-performance liquid chromatography. The data set was studied by means of chemometric methods. Partial least squares regression was used to obtain models that show a significant correlation between composition of the oil’s polar compounds and conditions of the olives sampled. In particular, compounds with antioxidative activity were directly linked with the state of health of the olives. The models obtained allow tracing of the state of health of the olives sampled through analysis of the polar fraction of virgin olive oil with a high degree of accuracy, and thus prediction of the oil’s expected shelf life.

A new analytical method for fatty acid alkyl esters (FAAEs) determination by GC-MS in virgin olive oils is proposed. No sample preparation is required and FAAEs are directly thermo-desorbed and cryo-focalised in the cooled injector of a GC-MS (EI) instrument. The analytical conditions were optimized by Design of Experiment (DoE) techniques (an exploratory Plackett-Burman design followed by a factorial design on three selected variables). After the improvement of method performances, several samples of extra virgin and low quality virgin olive oils were analyzed both by the new method and by the Official EU Method of analysis. The application of Principal Component Analysis to the obtained results confirmed that the ability of the proposed method to discriminate between extra virgin and lower quality olive oils is at least equal to that of the Official Method, but the new method is faster, simpler, requires a much lower amount of organic solvents and significantly enhances method repeatability.

Two methods for routine analysis of free 7-ketocholesterol (7-k) in foods by high-performance liquid chromatography (HPLC) were compared. The analyzed samples were egg noodles, biscuits prepared with eggs, sweet snacks, grated Parmesan cheeses, and some ingredients utilized in the food industry (whole-milk and whole-egg powder). The enrichment of cholesterol oxides was carried out by solid-phase separation of the total lipids with florisil and silica cartridges for methods A and B, respectively. The 7-k analyses were run in normal-phase HPLC for method A, and a reverse-phase process was used in method B. Identification of the 7-k peak was confirmed by gas chromatography/mass spectrometry analysis and peak purity checkvia spectral analysis with a diode array detector. The quantitation of 7-k was carried out with an internal standard for method A and with a calibration curve for method B. The limit of quantitation (LQ) was 3 × 10−9 g/injection; the limit of detection was ten times lower than the LQ for both methods. The two methods showed good recovery (99%) and good repeatability (coefficient of variation of 3.9 and 3.7% for methods A and B, respectively). These methods allow a fast, sensitive, and reliable determination of one cholesterol oxidation product, which is present at a high concentration level in the first stages of the oxidation process.

Consumer demand for pomegranate juice has considerably grown, during the last years, for its potential health benefits. Since it is an expensive functional food, cheaper fruit juices addition (i.e., grape and apple juices) or its simple dilution, or polyphenols subtraction are deceptively used. At present, time-consuming analyses are used to control the quality of this product. Furthermore these analyses are expensive and require well-trained analysts. Thus, the purpose of this study was to propose a high-speed and easy-to-use shortcut. Based on UV–VIS spectroscopy and chemometrics, a screening method is proposed to quickly screening some common fillers of pomegranate juice that could decrease the antiradical scavenging capacity of pure products. The analytical method was applied to laboratory prepared juices, to commercial juices and to representative experimental mixtures at different levels of water and filler juices. The outcomes were evaluated by means of multivariate exploratory analysis. The results indicate that the proposed strategy can be a useful screening tool to assess addition of filler juices and water to pomegranate juices.

The aim of this study was to evaluate the effect of the addition of carnosic acid on the oxidative stability of virgin olive oil. Two different amounts of carnosic acid (0.01 and 0.1 g/100 g oil) and two different temperatures (accelerated aging temperature, 60 °C; deep frying temperature, 180 °C) were considered. The influence of carnosic acid and heating time on the stability of the oils was studied by experimental design. The results obtained at 60 °C showed a dose dependent inhibition in the formation of primary and secondary oxidation products and a dose dependent enhancement of radical scavenging activity, which was only less significantly influenced by heating time. On the contrary, at 180 °C no protective effect against lipid oxidation was observed and the radical scavenging activity was practically zeroed by heating, probably as a consequence of a fast decomposition of carnosic acid.

NIR spectroscopy is proposed as a rapid and effective analytical method for the identification of the cultivar of table olives. The study was particularly focused on the Taggiasca cultivar. A set of 46 samples representative of the Taggiasca production area was analysed together with 43 samples of table olives of different cultivars. After feature selection, LDA and SIMCA were applied to the NIR data as classification and class-modelling techniques, respectively. The excellent results obtained by NIR spectroscopy (mean specificity of the models 82.0%, sensibility >90%) were then compared to those obtained by the gas chromatographic (GC) determination of the fatty acids composition of the oils extracted from the samples of table olives. Finally, in order to test the potential synergy among different sources of information, the employed NIR and chemical variables were joined, but only a small improvement of the results obtained by NIR alone was reached.

The volatile fraction of different kinds of Pesto genovese, a typical Italian basil-based pasta sauce, was analysed by Headspace Sorptive Extraction (HSSE)-Gas Chromatography–Mass Spectrometry (GC–MS) coupled with chemometric methods. Three kinds of commercial samples were considered: (a) non heat-processed shop manufactured, (b) heat-processed industrial and (c) non heat-processed industrial samples. The category of heat-processed samples was easily discriminated by the amounts of the terpene hydrocarbon α-terpinene. In order to distinguish the three categories, multivariate statistical analysis was then performed. Good prediction results were obtained by the combination of Linear Discriminant Analysis (LDA) and Stepwise LDA (STEPLDA): the percentage of correct predictions was 92 for food industry manufactured non heat-processed samples and 100 both for shop manufactured, non heat-processed samples, and for food industry manufactured, heat-processed samples.

Cross-flow microfiltration (MF) and ultrafiltration (UF) with different commercial membranes were applied to virgin olive oil (VOO) in order to remove some compounds that are responsible for oil unpleasant flavor without altering the oil composition. Experimental tests were carried out using a Spanish VOO from Andalusia, which had the typical eucalyptus aroma. Several classical analytical parameters (acidity, peroxide number, UV absorbance) were determined. Moreover, the contents of total chlorophyll and minor polar compounds were detected. The effect of filtration on oil aroma was evaluated by detecting carbonyl compounds and by a panel of trained tasters. The UF Carbosep M1 membrane was the most suitable for softening the oil organoleptic features. Carbosep UF M1 and MF M14 membranes also induced a reduction of the content of total chlorophyll, which certainly slowed down the oil oxidation processes. It should be considered, however, that the observed decrease in the content of phenolic compounds with Carbosep M1 might have an adverse affect on the oil's stability to oxidation, but the observed decrease in total chlorophyll content should oppose it.

This study aimed at evaluating if the volatile terpenoid hydrocarbons of extravirgin olive oils from West Liguria, a North Italy region, could trace their geographical origin. If terpenoid hydrocarbons were individually considered, three compounds, i.e. alpha-copaene, alpha-muurolene and alpha-farnesene, allowed building a simple decision tree and discriminating oils produced in West Liguria from oils produced in other Mediterranean regions. Moreover, the multivariate analysis allowed building West Liguria class-models with high predictive ability, confirming the fundamental role of the volatile terpenoid hydrocarbons for the geographical characterisation of West Liguria oils.

The aim of this preliminary study was to assess the possible presence of cholesterol oxidation products in 2 i.v. lipidic emulsions with different fatty acid compositions (long-chain triglyceride, medium-chain triglyceride-long-chain triglyceride). Because these emulsions are currently used in neonatal parenteral nutrition, their direct venous introduction might be potentially dangerous because of the possible atherogenic role of cholesterol oxidation products. The emulsions were analyzed when bottles were opened (ie, under normal condition of administration) and after a 12-hour direct experimental exposure to air and high (90%) oxygen concentrations. 7-Ketocholesterol and 5alpha-epoxycholesterol were chosen as markers of cholesterol oxidation and detected by gas chromatography-mass spectrometry of their trimethylsilyl ethers. The detected amounts were always very low and in some cases below the detection limit of the analytical method for the 2 cholesterol oxidation products (COPs; 0.1 and 0.3 microg/g of extracted lipids). Immediately after opening the bottles, their concentrations were lower in the emulsions containing the higher amounts of polyunsaturated fatty acids. Experimental hyperoxic exposure generally determined only a mild increase in the content of cholesterol oxidation biomarker, and after exposure to oxygen, the amounts of COPs were slightly higher than after exposure to air. The results of the present study are undoubtedly reassuring for the safety of neonates, although caution is always required when drawing conclusions from in vitro data.

The quantity of blocked lysine in spray-dried infant fistulas in Italy was calculated by the furosine method. This quantity was then correlated with the composition of the carbohydrate fraction. In formulas containing only lactose as carbohydrate its presence in large quantities (6.7-74.5 g/100 mL of formula as fed), with a lysine-rich protein fraction, may lead to amino acid blocking of >20%. In formulas in which lactose was substituted with low-DE maltodextrin a reduction of lysine blocking was possible. Addition of glucose may result in high levels of blocked lysine

In the last few years, a variety of experimental and clinical studies concerning the formation, metabolism, and cellular effects of cholesterol oxidation products (COPs) have been carried out. Nevertheless, a substantial lack of knowledge exists regarding the possible intake of these compounds by the newborn through human and/or adapted formula milk. As far as the pathological role of COPs is concerned, exhaustive studies have shown that since dietary COPs are cytotoxic and atherotoxic, they may lead to adverse effects on health. The aim of this study was to investigate the possible development of cholesterol oxidation in adapted formula and in human milk by comparing the main cholesterol oxidation biomarker (7-ketocholesterol) concentration in both. To do so, the total (bonded and free) 7-ketocholesterol content was measured in ten fresh human mature milk samples and in ten milk adapted formula samples by high performance liquid chromatography and diode array detection. The 7-ketocholesterol human milk content (0.7±0.3) was often below the quantifiable limit (0.5μg/g of extracted lipids) while 7-ketocholesterol adapted milk concentrations were often above (3.6±4.0) this limit. The 7-ketocholesterol content of adapted formula samples was significantly higher as compared to human milk samples (P This is the first study to provide data concerning the concentrations of cholesterol oxides in human milk and in formula milk. Our results clearly suggest that the manufacturing technologies employed and the nutrient extractive sources play a crucial role in the development of cholesterol oxides in the end product. Careful surveillance has to be paid in order to avoid alteration of bioactive properties of nutrients and/or development of potentially toxic derivative compounds.

7-Ketocholesterol and 7-ketositosterol were chosen as reliable markers of the oxidation of cholesterol and phytosterols in infant milk formulas and infant milk cereals. A reversed-phase HPLC method was developed to measure them simultaneously in infant formulas. This method was then tested on a wide range of infant milk formulas and milk cereals on sale in Italy whose lipid composition is representative of the most common commercial formulas. The analytical results revealed no significant differences in the extent of oxidation of cholesterol and sitosterol. As the level of 7-ketocholesterol often followed the cholesterol level, a cholesterol content similar to that of human milk produced amounts of cholesterol oxides with possible negative effects on infant health. In contrast, the low cholesterol content of milk cereals never produced amounts of cholesterol oxides high enough to cause concern. The contents of phytosterols and hence their oxides were always low.

The degree of cholesterol oxidation in commercial sweet baked foods (biscuits and snacks) and in laboratory baked biscuits, all containing fresh or powdered eggs, was determined. 7-Ketocholesterol was used as index of cholesterol oxidation and detected by two analytical methods. The analysis of the biscuits showed higher levels of 7-ketocholesterol and a more marked oxidative instability of cholesterol when prepared with powdered eggs. The significant amounts of 7-ketocholesterol found in some samples of commercial biscuits were attributed to the use of powdered eggs. These data are of importance to industries using eggs in sweet baked products which are mainly consumed by children.

Sterol oxidation was evaluated in commercial meat- and fish-based homogenized baby foods containing vegetable oil. Gas chromatography coupled with mass spectrometry (GC/MS) was used for the analytical determination of 7-ketocholesterol and 7-ketositosterol, which were chosen as markers of sterol oxidation in lipids of animal origin and vegetable origin, respectively. Cholestanetriol was also quantified, because its negative effects on atherogenesis and other biological processes are well known. In meat-based samplesg/serving. The 7-ketocholesterol and 7-ketositosterol were 2289 and 1140 g/serving, respectively, whereas the 7-ketocholesterol/cholesterol percent ratio was and 11-40 g/serving, respectively, whereas the cholestanetriol levels were 738 g/serving. The 7-ketocholesterol/cholesterol percent ratio was characteristic of each kind of meat and related to the levels of unsaturated fatty acids of animal lipids. In fish-based samples, the cholestanetriol and 7-ketocholesterol levels per serving were significantly lower than in meat samples, but in fish fillets they were about 2025, instead of 40, of the ingredients. The values of the 7-ketocholesterol/ cholesterol percent ratio in fish-based products were close to the values computed for chicken or turkey-based products. The detected values of cholestanetriol showed that the addition of vegetable oil enhances the development of the indirect bimolecular pathway of cholesterol oxidation, which was even more favorable in fish-based products.