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We have investigated the incorporation of cholesterol oxidation products into cardiomyocyte lipids and related this to changes in cell proliferation, evaluated by measuring cellular protein content. Primary cultures of neonatal rat ventricular cells were supplemented with scalar concentrations of several cholesterol oxidation products (cholestan-5 alpha, 6 alpha-epoxy-3 beta-ol, 5 alpha-cholestane-3 beta, 5, 6 beta-triol, 5-cholesten-3 beta, 4 beta-diol, 5-cholesten-3 beta-ol-7-one, and 5-cholesten-3-one). Although all the cholesterol oxidation products were incorporated into the cardiomyocyte lipids when added to the medium at a concentration higher than 0.5 microM, the extent of the incorporation of the different cholesterol oxidation products differed, depending on the concentration in the culture medium and on the chemical structure of the compound. The effects of the cholesterol oxidation products on the cellular protein content were also different: 5 alpha-cholestane-3 beta, 5, 6 beta-triol was shown to be the most potent inhibitor of cell proliferation, followed by cholestan-5 alpha, 6 alpha-epoxy-3 beta-ol, 5-cholesten-3 beta, 4 beta-diol and 5-cholesten-3 beta-ol-7-one. 5-Cholesten-3-one did not affect the cellular protein content. The ability of cholesterol oxidation products to inhibit cell proliferation, and their capacity to increase the permeability of the plasma membrane to calcium, could be deleterious for cardiac cells.

In order to evaluate the effect of one of the main oxysterols derived from cholesterol oxidation, cholesterol-5 alpha,6 alpha-epoxide (epox), on cardiac cells, we have supplemented the culture medium of neonatal rat cardiomyocytes with scalar concentrations of epox (0.1-100 microM). While 0.1 microM epox supplementation was ineffective, epox supplementation in the range 1-100 microM determined a reduction in cellular protein level, without affecting cell viability, and a dose-dependent epox incorporation into cardiomyocyte lipids. Furthermore, in the same concentration range of epox supplementation, a gas chromatographic peak unambiguously identified by gas chromatography-mass spectrometry as cholestane-3 beta,5 alpha,6 beta-triol, an hydrolytic metabolite of epox, was detected. The mechanism of cytotoxicity of epox to cardiomyocytes could be due to the insertion of epox itself into cellular lipids, and to its metabolization to the more toxic triol.

Parmigiano–Reggiano (P–R) cheese is still one of the most valuable Protected Designation of Origin (PDO) cheeses of Italy. The rind percentage (from 12% to 50%), the ripening (from 13 to 31 months), the moisture content and the differences between true P–R and competitors were determined by means waveguide spectroscopy. Preliminary tests were carried out in the 2–20 GHz frequency range (with a span of 1 GHz) to investigate which 1 GHz frequency range contains most information on the rind percentage and on the months of ripening. Partial Least Squares (PLS) regression was used to predict rind percentage, months of ripening and moisture in the previously selected frequency ranges (2–3, 5–6 and 17–18 GHz for the rind percentage; 2–3, 5–6 and 16–17 GHz for the months of ripening). Moreover, Soft Independent Modelling of Class Analogy (SIMCA) analysis was used to discriminate the samples according to the rind percentage. Principal component analysis (PCA) was used to discriminate true P–R cheese from competitors. PLS models ( test set validation ) showed R 2 values up to 0.944 (root mean square error of prediction in test set validation, RMSE p = 3.4%), 0.966 (RMSE p = 1.2 months) and 0.786 (RMSE p = 0.99%) for the prediction of rind percentage, months of ripening and moisture percentage, respectively. As a result only a part of the prediction of rind percentage and ripening can easily be attributed to the moisture. For each considered frequency range, all samples belonging to the classes characterized by 12% and 50% of rind were correctly classified. Competitors were clearly separated from P–R cheese by the PCA analysis of the spectral signals and their moisture was predicted with a R 2 values up to 0.942 ( test set validation ). In this case, the discrimination power can be mainly attributed to moisture content.

Rapid analytical techniques were used to analyze the lipid fraction and to characterize phenolic compounds in Italian walnuts (Chandler cv.) grown in the same experimental orchard under different agronomical conditions. Lipid components such as fatty acids, sterols and tocopherols were determined by capillary gas chromatography using a flame ionization detector. Linoleic and linolenic acids were the most representative ω-6 and ω-3 essential dietary fatty acids present. A total lipid chromatogram was used to simultaneously determine free sterols and tocopherols. Walnuts contained substantial amounts of γ- and δ-tocopherol, which explains their antioxidant properties. Sitosterol, Δ5-avenasterol and campesterol were the major free sterols found. Micellar electrokinetic chromatography was used to analyze simple and complex phenols (tannin profile), while the total phenol amount and the antioxidant activity of phenolic extracts were assessed by spectrophotometric assays with Folin-Ciocalteu and ABTS•+ reagents, respectively. The total phenol content was higher than most foodstuffs. Moreover, more than 96% of the phenolic fraction belonged to the tannin class. Statistical analysis of the data showed that higher levels of nitrogen fertilization led to a significant increase in linoleic acid. This effect can induce a faster oxidation of lipid fraction of walnuts. Nevertheless, major nitrogen fertilization and irrigation were able to enrich walnuts in phenolic compounds. PRACTICAL APPLICATIONS Walnuts (Juglans regia L.) are a good source of essential fatty acids, tocopherols and phenolic compounds that contribute to reducing the risk of coronary heart disease, and also inhibit the oxidation of human plasma and low density lipoproteins. In this work, the applicability of fast analytical determinations was utilized to discriminate different walnut samples. Total lipid chromatogram was used for the simultaneous determination of the profile of sterols and tocopherols; micellar electrokinetic chromatography was used for the phenolic separation, which takes less than 10 min. The proposed methods are ideal for routine analysis of the lipidic and phenolic fractions in order to control the quality of walnuts.

The effect of the fluorescent light exposure and type of packaging (normal atmosphere and oxygen-rich atmosphere) was evaluated on the oxidation parameters (peroxides and cholesterol oxidation products) of raw beef slices placed in packed vessels and refrigerated. The concentration of COPs in meat treated under modified atmosphere ranged from 0.15 to 0.52 mg/100 g meat (average value of 0.27 mg COPs/100 g meat), which was twice as much the average COPs content (0.14 mg/100 g) of meat packed under air (0.04 to 0.27 mg COPs/100 g meat). The main cholesterol oxide was 7k, which represented about one third of the total cholesterol oxides, followed by 7beta-OH (20-25% of total COPs), 7alfa-OH (about 20%) and beta-epoxy (12-18%). In normal atmosphere, photoxidation was a superficial process, since an inverse correlation between meat slice weight and COPs content on lipid basis was observed, unlike in high-oxygen (32%) atmosphere.

The aim of this work was to develop a capillary electrophoresis-mass spectrometry (CE-MS) method to identify and quantify phenolic and other related polar compounds in walnut samples. The extraction capacity of several solvent mixtures of phenolic compounds from walnut by conventional solid-liquid extractions was tested, and CE and electrospray ionization MS parameters were optimized. The finalized procedure is able to determine many well-known phenolic compounds present in walnuts and provide relevant information about the presence of minor polar compounds. A new compound in walnut ((2E,4E)-8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-beta-d-glucopiranosyl ester, [M-H](-) 403.161m/z) with a structure similar to glansreginins was also identified. Phenolic compounds correspond to 14-28% of total polar compounds quantified. Aglycone and glycosylated ellagic acid represent the principal components and account for 64-75% of total phenols in walnuts. However, the sum of glansreginins A, B and (2E,4E)-8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6'-O-beta-d-glucopiranosyl ester was in the range of 72-86% of total quantified compounds. In addition, this is the first time that separation by CE with detection by electrospray ionization time-of-flight MS has been applied to the analysis of phenolic and other polar compounds in walnut samples, providing results in less than 15min.

A new CZE method was developed and compared with HPLC for the determination of (E)-10-hydroxy-2-decenoic acid (10-HDA) in royal jelly (RJ) samples of different geographical origin. The results obtained with the CZE method were highly correlated with those of HPLC (p < 0.01). Under optimized conditions, CZE employed minimal amounts of 50 mM tetraborate buffer as BGE, without the addition of organic solvents, EOF or pH modifiers. The CZE method showed a wide linear response range (0.006-0.808 mg 10-HDA/mL), a good sensitivity (LOD and LOQ were 0.002 and 0.004 mg/mL, respectively) and a satisfactory instrumental repeatability with respect to migration time and peak area (RSD% less than 1.0 and 2.0% on migration time for intra- and interday assay, respectively and less than 2.0 and for 4.0% on peak area for intra- and interday assay, respectively). The 10-HDA content in RJ ranged from 0.8 to 3.2 g/100 g of RJ and a significant difference (p < 0.05) was found between the Italian and extra-European average values: 2.5 and 1.6 g/100 g of RJ, respectively, according to the CZE data. The possibility of application of CZE for routine analyses on RJ and RJ based products to verify their authenticity is highlighted here.

The oxidative stability of egg yolks from brown hen eggs, previously subjected to a 6-week storage at room temperature, was evaluated as related to the dietary supplementation. Three groups of eggs yolks were obtained according to the carotenoid supplementation: no addition of apo-ester and canthaxanthin (CON); 2.5 ppm of apo-ester and 2.5 ppm of canthaxanthin (MED); 3.5 ppm of apo-ester and 5.5 ppm of canthaxanthin (HIGH). The lipid content of the egg yolks varied from 28.3% to 29.3%, and no significant differences were found among the different dietary treatments (p≥0.05). Peroxide value (PV) ranged from 0.92 (CON) to 1.48 (HIGH) meq O2/kg lipids, which were quite low despite the 6-week egg storage at room temperature. The most common cholesterol oxidation products (COPs) in foods were detected in the stored eggs. The total COPs content varied from 28.2 (CON) to 39.2 (HIGH) µg/g lipids, which corresponded to 8.32 and 10.97 µg/g egg yolk, respectively; however, no significant differences (p≥0.05) on the total COPs level were observed among the different dietary treatments. Regarding the single COPs, the most abundant was 7α-hydroxycholesterol (50-53% of total COPs), followed by 7-ketocholesterol (22-26%), 5,6β-epoxycholesterol (7-9%), 7β-hydroxycholesterol (6-9%), 5,6α-epoxycholesterol (3-4%), and cholestanetriol (2-4%). In conclusion, PV and total COP content of egg yolks confirm that the use of dietary supplementation with carotenoids is not a useful strategy to control and delay the lipid autoxidation in brown hen eggs stored for 6 weeks at room temperature.

Background and objectives: The high nutritional value and low cost of soybean products make them a suitable solution for malnutrition problems for poor people living on grain centred diets. Tofu, a fundamental part of Asian food culture, is a traditional oriental soybean food and it may be the most popular food made of soy worldwide. Thousands of studies (in vivo and in vitro, with animals and human subjects) have shown that soybeans and soy components have many health-promoting effects; some of them were attributed to the isoflavones. In particular, various forms of isoflavones have been shown to possess different antioxidant activities. The aim of the study was to investigate the effect of lactic acid bacteria (LAB) fermentation on the free and bound phenolic fraction of tofu. Methods: The fermentation process was established using specific strains of lactic acid bacteria. Free and bound phenolic compounds from soybean, traditional tofu and tofu obtained by soymilk fermentation were studied by HPLC-DAD-ESI-MS. Results: Important effects of fermentation were seen on isoflavone profile: in fact, isoflavones content of fermented tofu showed significant differences compared to soybean and to traditional tofu. In particular, fifteen isoflavones were identified as aglycones and glycosides: the aglycone forms were significantly higher in fermented tofu compared to the traditional one. The ratio of aglycones/glycosylated isoflavones was 0.34 and 2.55 for the traditional and fermented tofu, respectively. Soy and traditional tofu presented the same content of bound phenolic compounds (mainly phenolic acids), instead fermented tofu showed a bound phenolic content 34 % higher than traditional tofu. Conclusions: These results suggest that fermentation by LAB plays an essential role in the increase of the content of aglycone isoflavones and bound phenolic compounds with important biologically implications on human health.

Phospholipids (PLs) are the most important class of polar lipids in foods because they are the major components of cell membranes in animal and plant tissues. Recent studies have shown that PLs can have a positive nutritional effect on human health, such as reducing the risk of cardiovascular disease, reducing blood cholesterol levels and enhancing brain function [ ]. As an integral component of cell membranes, they are involved in cell signalling and, therefore, they are indispensable for the communication and interaction between body cells [ - ]. Although some influences of phospholipids on health are controversial, overall dietary phospholipids are most likely to have a positive effect on health [ ]. Nevertheless, phospholipids are poorly investigated as minor compounds in olive oil and in literature the phospholipids extraction has been essentially developed by liquid-liquid extraction (LLE). The aim of this work was to compare the effectiveness of a previous LLE extraction method with a direct separation and purification of olive oil phospholipids with two different SPE sorbents (silica and diol). As far we are concerned this is the first application of SPE for the direct extraction of phospholipids in virgin olive oil. Furthermore, the extracts were analyzed by HPLC-ESI-qTOF-MS to obtain basic information useful for a possible identification of the single components. To this aim, a new separation method was established using an hydrophilic fused core liquid chromatography column (HILIC). The HILIC chromatography is readily compatible with electrospray ionization and results in higher sensitivity and improved peak shapes. The SPE diol extraction and HPLC-ESI-TOF-MS methodology have demonstrated to be very useful to determine the phospholipids in not filtered virgin olive oil and also in a veiled virgin olive oil subjected to a glyceridic crystallization due to a temperature decrease. In fact, it could be observed that crystallization caused a significant decrease in the phospholipids fraction.

Background and objectives: According to literature the whole cold-pressed lipid fraction extracted from pomegranate seeds, if assumed through diet, can improve immune function in vivo, reduce hepatic triacylglycerol accumulation and act as a chemopreventive agent against hormone-related human cancers. Pomegranate seed oil comprises 10–20% of total seed weight, with an ideal highly unsaturated fatty acid content, mainly constituted by the rare conjugate linolenic acid isomer punicic acid, which activities against inflammation and metabolic syndrome have been confirmed by various in vivo experiments. However, besides several very encouraging studies on the possible beneficial effects of this oil, little is known about the composition of its minor fraction. Therefore, in this work, the main lipidic classes of 17 pomegranate varieties were determined. Methods: The pomegranate oil was extracted by Folch method and several chromatographic techniques were used to determine fatty acid, tocopherol, sterol and phospholipid composition. Results: The total lipid content of pomegranate seeds varied between 7.9 and 16 %. Pomegranate seed oil contained higher amount of CLnA (punicic acid) in the range of 73.6-85.0 % of total fatty acid content. Other major fatty acids were oleic, linoleic and palmitic acid, respectively. Pomegranate seed had high contents of tocopherol (0.6-2.6 mg/g of fat). Delta-tocopherol was 91.2-96.7 % of total tocopherols. The major phytosterols analyzed were delta5-avenasterol, campesterol, stigmasterol, and â-sitosterol. Moreover some triterpene compounds as betulin, botulino and cyclolanostanol were identified. Three different phospholipids were detected; phosphatidylethanolamine was the first compounds in the range of 44-91% of total phospholipids. Phosphatidylinositol and phosphatidylcholine were, also, quantified. Conclusions: In summary, these data indicated that pomegranate seeds are rich sources of CLnAs, tocopherols, phospholipids and sterols with potential beneficial physiological activities. In addition, different pomegranate cultivars may differ in their seed components and health properties.

Background and objectives: Proanthocyanidins are mixtures of flavan-3-ol units linked mainly via B-type bonds, i.e., C4-C8 or C4-C6 linkages. Occasionally an additional C2-O7 or C2-O5 linkage may exist, leading to doubly bonded A-type. These phenolic compounds are widely present in plants and have been suggested to demonstrate antioxidant, antibacterial, antiviral, anticarcinogenic, anti-inflammatory, and vasodilatory activities. Surprisingly, quantitative information on the proanthocyanidin profiles in plant products is quite lacking, largely due to the absence of appropriate analytical methodology and commercially available standards for oligomers. The study reported here was undertaken to detect and characterise the procyanidins in different type of barley. Methods: Fourteen barley varieties, four waxy and ten no-waxy, were collected and characterised. Proanthocianidin compounds were extracted and their separation was perfomed by normal phase HPLC-FLD-MS. Results: Monomers, procyanidin and prodelphinidin dimers, trimers, tetramers, and procyanidin pentamers were identified by mass spectrometry and quantified by fluorimetric detection. The total content of proanthocyanidins in barley samples ranged between 29.3 to 65.3 mg/100g. In all samples, the principal components consisting of procyanidin and prodelphinidin trimers with structure (gallocatechin-catechincatechin), followed by the procyanidin dimer and representing on average 19, 17 and 16 % of the total content. With exception of some barleys, monomer content was low. Waxy barley varieties reported the higher content of phenolic compounds compared to no-waxy varieties. Conclusions: These results estabilished that the barley varieties analysed in this work contain proanthocyanidins from monomer to pentamers and how these oligomers are a mixture of procyanidins and prodelphinidins, consisting mainly of catechin units. Moreover the waxy varieties showed the higher content of proanthocyanidins

The fatty acid profile of 32 butter samples were analyzed by fast gas-chromatography. Italian and foreign butters, 21 and 7 respectively, were collected at local markets in Europe, while 4 experimental butter samples were produced at a local dairy farm in the Parmigiano-Reggiano Cheese Consortium (CFPR) area, from fresh outcrop milk creams obtained by supplementing the traditional diet of dairy cows with 300 g/d of extruded linseed. With the respect to the Italian and foreign butter samples, experimental butter showed a lower content of medium-chain SFA (-8.5% and -8.6%), a better SFA/UFA ratio (1.78), a higher content of CLA (+32% and +15,3%), total n-3 PUFA (+56,4% and +28,8%) and trans fatty acids (+23.8% and +11.1%). This study support the feasibility for a sustainable production of value-added by-product of dairy industry, by improving cow’s feeding strategy to achieve a higher content of bioactive fatty acids in milk fat, leading to the production of dairy products that may be more beneficial to consumers.

Background and objectives: Pomegranate (Punica granatum) has gained widespread popularity as potential functional food. In different studies, pomegranate juice (PJ) has been shown to exert antiatherogenic, anti-inflammatory, and antioxidant effects, which have been attributed to its high phenolics content. The differences in polyphenols contents and antioxidant activities of commercial PJ, related to cultivar and/or industrial processing [1], must be carefully taken into accounts in evaluating the health effects of PJ. The aim of this study was to rank PJ from different cultivar according to their total phenolic content and antioxidant capacity. According to the ranking three PJ (Wonderful, Hicaz and G2) were then chosen, characterized for their phenolic composition, and used for HepG2 cells supplementation to evaluate their biological protective activity. Methods: Cells were supplemented with PJ at two different medium concentration, and an oxidative stress was determined in some experiments by cell exposure to H2O2. Results obtained in PJ supplemented cells were compared to unsupplemented controls (stressed and not stressed) and to cells supplemented with tocopherol. Results: In stressed condition, all PJ but Hicaz at the highest concentration counteracted H2O2 induced cytotoxicity. Furthermore, the oxidative damage was effectively counteracted by all PJ in a dose and cultivar-dependent manner. Conclusions: Our results point out the importance of the cultivar in the degree of protection that can be achieved in cells after PJ supplementation, in some cases comparable to vitamin E.

Sample preparation is an important step for the determination of phenolic compounds in biological samples. Different extraction methods have been tested to determine phenolic compounds and their metabolites in plasma by nano-liquid chromatography coupled to electrospray ionization-time of flight mass spectrometry (nanoLC-ESI-TOF-MS). The sample treatment optimisation was performed using commercial foetal bovine serum spiked with representative phenolic standards, namely naringenin, luteolin, verbascoside, apigenin, rutin, syringic acid and catechin. Different protein-precipitation conditions were evaluated as well as enzymatic digestion with trypsin and solid-phase extraction using different phases such as C-18, ABN and ENV+, working at different pH values. The optimum extraction procedure consisted of a previous protein-precipitation step using HCl 200 mmol/L in methanol for 2.5 h at 50ºC followed by a solid-phase extraction using C-18 cartridges at pH 2.5. This procedure was finally applied to the plasma of rats overfed with a phenolic-rich Lippia citriodora extract. These samples were analysed by nanoLC-ESI-TOF-MS, enabling the identification of five compounds previously found in the administered Lippia citriodora extract and one metabolite.

Soy foods have long been a staple of the human diet in Asia and tofu may be the most popular food made of soy worldwide. Tofu is prepared from soymilk curd and it can also be further processed (grilled, frozen, or fermented) modifying taste, texture and end-uses compared to traditional tofu. Fermentation also prevent undesired microbial and chemical reactions improving the stability and the quality of the product. Moreover, in addition to be natural, nutritious and safe, epidemiological studies have shown that fermented soybean products exhibit healthy effects. In fact, soybean and its products contains biologically active compounds including isoflavones, essential fatty acids, sterols, phospholipids and tocopherols which may contribute to prevent chronic diseases such as hormone-dependent cancers, cardiovascular diseases and osteoporosis. Due to the influence of processing on the bioactive compounds content of soy food products, soybean, traditional tofu and tofu obtained by soymilk fermentation were studied by GC–FID and HPLC coupled with fluorimetric, light scattering and electrospray ionization mass spectrometry detector, in order to analyse their lipidic profile (fatty acids, tocopherols and sterols) and isoflavones content and composition. In particular, for isoflavones analysis, a new method was set up. Fermentation was performed using specific strains of lactic acid bacteria (LAB), such as Lactobacillus casei and Lactobacillus acidophilus. Fatty acids profile of soybean and tofu samples were very similar, even in term of trans fatty acids and omega-3 fatty acids, so fermentation cannot differentiate tofu from soybean by fatty acid composition. The most abundant fatty acid was linoleic acid, followed by oleic acid, palmitic acid and α-linolenic acid, as reported by other authors. The processing conditions did not significantly affect neither tocopherol composition of tofu (traditional and fermented) compared to soybean and in all products the main tocopherol was γ-tocopherol followed by δ-tocopherol and α- tocopherol. As concerning phytosterol content, ß-sitosterol was the main sterol found with the phytosterol composition that was similar in soybean and traditional and fermented tofu. The main effects of fermentation were on isoflavone profile: in fact, significantly differences were seen in fermented tofu isoflavones content compared to soybean and to traditional tofu. In particular, fifteen isoflavones were identified as aglycones and glycosides: the aglycone forms were significantly higher in fermented tofu compared to the traditional one. These results suggest that fermentation by LAB cannot discriminate the fatty acid, tocopherol and phytosterol composition of fermented tofu from the traditional one and from soybean, but it plays an essential role not only on shelf-life extend, but also in the increase of the content of aglycone isoflavones with important biologically implications on human health.

Barley byproducts obtained by air classification have been used to produce a different barley functional spaghetti, which were compared to different commercial whole semolina samples. Total, insoluble, and soluble fiber and β-glucan contents of the barley spaghetti were found to be greater than those of commercial samples. Furthermore, it was proved that barley spaghetti reached the FDA requirements, which could allow these pastas to deserve the health claims “good source of dietary fiber” and “may reduce the risk of heart disease”. When the barley coarse fraction was used, a flavan-3-ols enrichment and an increase of antioxidant activity were reported, while commercial samples showed the absence of flavan-3-ols and a higher presence of phenolic acids and tannins. Whole semolina commercial spaghetti had a significantly higher content of phenolic acids than semolina spaghetti samples. Besides, it was observed that when vital gluten was added to the spaghetti formulation, phenolic compounds were blocked in the gluten network and were partially released during the cooking process.

Nowadays there is considerable interest in the consumption of alternative crops as potential recipes for gluten-free products production. Therefore, the use of buckwheat for the production of gluten-free pasta has been investigated in the present study. RP-HPLC-ESI-TOF-MS has been applied for the separation and characterization of free and bound phenolic compounds in buckwheat flour and buckwheat spaghetti. Thus, 32 free and 24 bound phenolic compounds in buckwheat flour and spaghetti have been characterized and quantified. To the authors' knowledge, protochatechuic-4-O-glucoside acid and procyanidin A have been detected in buckwheat for the first time. The results have demonstrated a decrease of total free phenolic compounds from farm to fork (from flour to cooked spaghetti) of about 74.5%, with a range between 55.3 and 100%, for individual compounds. The decrease in bound phenols was 80.9%, with a range between 46.2 and 100%. The spaghetti-making process and the cooking caused losses of 46.1 and 49.4% of total phenolic compounds, respectively. Of the total phenolic compounds present in dried spaghetti, 11.6% were dissolved in water after cooking.

The reduced risk of chronic diseases related to whole grain consumption is in part attributed to their high antioxidant content. Many studies have been performed on the in vitro antioxidant capacity of cereals, but in vivo studies are necessary. We have evaluated and compared the effect of whole grain durum wheat bread and whole grain Kamut® khorasan bread on the oxidative status in rats. Two different bread-making processes were used for whole grain Kamut® khorasan, sourdough and baker’s yeast. After 7 weeks on the experimental diets rats were divided into two subgroups, one receiving an oxidative stress by doxorubicin injection. Our results evidenced both wheat durum and Kamut® khorasan as good sources of antioxidants, and a lower oxidative state in rats fed the cereal-based diets. Furthermore, Kamut® khorasan bread fed animals had a better response to stress than wheat durum fed, especially when a sourdough bread was supplied. Although further studies are needed, data herein reported suggest whole grains, particularly whole ancient grains, as a safe and convenient way of increasing antioxidant protection.

Apigenin and apigenin glycosides are the main flavonoids in artichokes. Thus, their 24 h-urinary excretion was investigated, as a marker of absorption, in six women after consumption of two steamed artichokes. No enzymes treatment was performed and the urine metabolites were investigated by HPLC-MS/MS. After a single dose of steamed artichokes, 0.76% and 0.16% of apigenin-monoglucuronide and apigenin, respectively, were recovered in urine collected during the first twelve hours. In the next 12 h, only 0.11 % and 0.02% of apigenin-monoglucuronide and apigenin were recovered. Our results suggest that apigenin and derivatives are absorbed in humans after consumption of steamed artichokes and their excretion into urine mainly occurs during the first twelve hours. In the present study it was not possible to verify the presence of apigenin-sulphate derivatives. Reference: [1] Pandino G, Courts FL, Lombardo S, Mauromicale G, Williamson G. Caffeoylquinic Acids and Flavonoids in the Immature Inflorescence of Globe Artichoke, Wild Cardoon, and Cultivated Cardoon. J Agric Food Chem 2010; 58:1026–31.

I risultati ottenuti relativi alla composizione in acidi grassi di oltre 170 campioni di burro e panne, provenienti dal Comprensorio di Produzione del Parmigiano Reggiano (CFPR) ed analizzati in questa ricerca, vengono confrontati con quelli ottenuti da Strocchi e collaboratori nel 1967, presso l’Istituto di Industrie Agrarie dell’Università di Bologna, su oltre 200 campioni di burri emiliani, gran parte dei quali provenivano da caseifici compresi nel CFPR.

The effect of the type of packaging film (transparent vs. light-protecting red film) was evaluated on the formation of cholesterol oxidation products (COPs) in refrigerated horse meat slices stored in retail conditions under light exposure for eight hours. In meat wrapped with a transparent film, COPs increased from 233 (control) to 317 μg/g of fat, whereas the red film delayed cholesterol oxidation and offered protection against COPs formation, since COPs decreased from 173 (control) to 139 μg/g of fat after 8 h of light exposure. In addition, light opened the epoxy ring and led to the formation of triol, which was actually absent at T0. A proper packaging film may represent a useful strategy to retard oxidative degradation in a light-sensitive, high pigment- and fat-containing food, such as horse meat.

Total phytosterols content was determined in the hexaploid wheat spp. Triticum aestivum (soft wheat) and T. spelta (spelt) and the tetraploid wheat spp. T. durum (durum wheat) and T. dicoccon (emmer wheat). Results showed significant quantitative differences between phytosterols content of tetraploid and hexaploid wheat species, and confirms the genetic affinity of T. aestivum with T. spelta and that of T. durum with T. dicoccon.

The effect of freeze-drying and the assessment of the storage stability of freeze-dried royal jelly were investigated by the determination of furosine and blocked lysine. The level of furosine in the royal jelly samples collected from cells at different times (1, 2 and 3 days after grafting) showed that the Maillard reaction had already occurred in the hive as indicated by the increase in furosine: from 9.6 to 20.8 mg/100 g protein. Freeze-drying increased the Maillard reaction slightly, since furosine content rose from 18.0 mg/100g protein of RJ harvested 3 days after grafting compared to 24.6 mg/100g protein of the lyophilised sample. Freeze-dried royal jelly was more prone to the early stage of the Maillard reaction than fresh RJ, as confirmed by the significantly higher furosine values found after 12 months, both at 4°C (253.4 versus 54.9 mg/100g protein) and at room temperature (884.3 versus 332.5 mg/100g protein). After 18 months at room temperature, the lyophilised samples reached a furosine level of 1440.4 mg/100 g protein, which corresponded to the blocked lysine levels, amounting to 24% of total lysine.

Background and objectives: Traditionally, pomegranate (Punica granatum L.) has been consumed as fresh fruit or as pomegranate juice and it has been described as a rich source of phenolic compounds. Several recent studies have shown that drinking pomegranate juice contributes to significant potential health benefits, and, today, it is known, that the positive health properties of pomegranate are due to the presence of bioactive compounds like, flavonoids, phenolic acids and tannins contained in it. Therefore, in this work, the main phenolic compounds of 17 pomegranate varieties were determined by HPLC-DAD-ESI-qTOF-MS. Methods: A new chromatographic HPLC-DAD-ESI-qTOFMS method, using a C18 fused-core column, was established and the analyses were performed in less than 20 minutes. Results: Eight anthocyanins and fourteen other phenolic compounds were tentatively identified and quantified in the pomegranate juices. As far we are concerned, a new flavonolglycoside has been tentatively identified for the first time in pomegranate juices. Total phenolic content ranged from 573.6 to 2519.8 mg/mL. Anthocyanins varied between 110.9 to 1925.5 mg/mL and they represented the 17-86 % of total phenolic content.Flavonoids were the 1.6-24.4 % of total phenolic compounds. Phenolic acid and ellagitannins were in the range of 300.9-810.2 mg/mL. Conclusions: The results confirmed, as reported in literature, that the pomegranate variety influences substantially the phenolic composition. It is important to highlight that, as far we are concerned, a new flavonol-glycoside has been tentatively identified for the first time in pomegranate juices thanks to this method.

The phenolic compounds in ten rocket salad samples were determined in this study using micellar electrokinetic chromatography (MEKC). A detailed method optimisation plan was performed to separate the phenols in rocket salad with the following four separation parameters: buffer concentration, buffer pH, applied voltage and temperature. The best results in terms of peak resolution and reproducibility among separations were obtained with a MEKC running buffer (pH 9.0) composed of 40 mM sodium tetraborate and 15 mM SDS. Using this new MEKC method, it was possible to separate the flavonoids in less than 5 min. The results obtained by MEKC analysis were compared with the data obtained by high-performance liquid chromatography (HPLC) coupled with DAD and MS detectors. The polyphenol study by HPLC-DAD-MS allowed the identification of two different classes of compounds. Quercetin and kaempferol derivatives were determined in the Diplotaxis and Eruca species, respectively. A positive correlation of phenolic compounds identified by MEKC and HPLC was found.