Questo lavoro di tesi è stato suddiviso in tre parti. L’argomento principale è stato lo “Studio della componente antiossidante di oli ottenuti da olive mediante l’utilizzo di diversi sistemi e parametri tecnologici”. E’ ben noto come la qualità ossidativa di un olio di oliva dipenda oltre che dalla sua composizione in acidi grassi, dalla presenza di composti caratterizzati da un elevata attività antiossidante, ovvero le sostanze fenoliche. I composti fenolici contribuiscono quindi in maniera preponderante alla shelf life dell’olio extravergine di oliva. Inoltre sono state riscontrate delle forti correlazione tra alcune di queste sostanze e gli attributi sensoriali positivi di amaro e piccante. E’ poi da sottolineare come il potere antiossidante dei composti fenolici degli oli vergini di oliva, sia stato negli ultimi anni oggetto di considerevole interesse, poiché correlato alla protezione da alcune patologie come ad esempio quelle vascolari, degenerative e tumorali. Il contenuto delle sostanze fenoliche negli oli di oliva dipende da diversi fattori: cultivar, metodo di coltivazione, grado di maturazione delle olive e ovviamente dalle operazioni tecnologiche poiché possono variare il quantitativo di questi composti estratto. Alla luce di quanto appena detto abbiamo valutato l’influenza dei fattori agronomici (metodi di agricoltura biologica, integrata e convenzionale) e tecnologici (riduzione della temperatura della materia prima, aggiunta di coadiuvanti in fase di frangitura e di gramolatura, confronto tra tre oli extravergini di oliva ottenuti mediante diversi sistemi tecnologici) sul contenuto in composti fenolici di oli edibili ottenuti da olive (paper 1-3-4). Oltre alle sostanze fenoliche, negli oli di oliva sono presenti altri composti caratterizzati da proprietà chimiche e nutrizionali, tra questi vi sono i fitosteroli, ovvero gli steroli tipici del mondo vegetale, che rappresentano la frazione dell’insaponificabile quantitativamente più importante dopo gli idrocarburi. La composizione quali-quantitativa degli steroli di un olio di oliva è una delle caratteristiche analitiche più importanti nella valutazione della sua genuinità; infatti la frazione sterolica è significativamente diversa in funzione dell’origine botanica e perciò viene utilizzata per distinguere tra di loro gli oli e le loro miscele. Il principale sterolo nell’olio di oliva è il β- sitosterolo, la presenza di questo composto in quantità inferiore al 90% è un indice approssimativo dell’aggiunta di un qualsiasi altro olio. Il β-sitosterolo è una sostanza importante dal punto di vista della salute, poiché si oppone all’assorbimento del colesterolo. Mentre in letteratura si trovano numerosi lavori relativi al potere antiossidante di una serie di composti presenti nell’olio vergine di oliva (i già citati polifenoli, ma anche carotenoidi e tocoferoli) e ricerche che dimostrano invece come altri composti possano promuovere l’ossidazione dei lipidi, per quanto riguarda il potere antiossidante degli steroli e dei 4- metilsteroli, vi sono ancora poche informazioni. Per questo è stata da noi valutata la composizione sterolica in oli extravergini di oliva ottenuti con diverse tecnologie di estrazione e l’influenza di questa sostanza sulla loro stabilità ossidativa (paper 2). E’ stato recentemente riportato in letteratura come lipidi cellulari evidenziati attraverso la spettroscopia di risonanza nucleare magnetica (NMR) rivestano una importanza strategica da un punto di vista funzionale e metabolico. Questi lipidi, da un lato un lato sono stati associati allo sviluppo di cellule neoplastiche maligne e alla morte cellulare, dall’altro sono risultati anche messaggeri di processi benigni quali l’attivazione e la proliferazione di un normale processo di crescita cellulare. Nell’ambito di questa ricerca è nata una collaborazione tra il Dipartimento di Biochimica “G. Moruzzi” ed il Dipartimento di Scienze degli Alimenti dell’Università di Bologna. Infatti, il gruppo di lipochimica del Dipartimento di Scienze degli Alimenti, a cui fa capo il Prof. Giovanni Lercker, da sempre si occupa dello studio delle frazioni lipidiche, mediante le principali tecniche cromatografiche. L’obiettivo di questa collaborazione è stato quello di caratterizzare la componente lipidica totale estratta dai tessuti renali umani sani e neoplastici, mediante l’utilizzo combinato di diverse tecniche analitiche: la risonanza magnetica nucleare (1H e 13C RMN), la cromatografia su strato sottile (TLC), la cromatografia liquida ad alta prestazione (HPLC) e la gas cromatografia (GC) (paper 5-6-7)

No description available.

This PhD thesis describes set up of technological models for obtaining high health value foods and ingredients that preserve the final product characteristics as well as enrich with nutritional components. In particular, the main object of my research has been Virgin Olive Oil (VOO) and its important antioxidant compounds which differentiate it from all other vegetables oils. It is well known how the qualitative and quantitative presence of phenolic molecules extracted from olives during oil production is fundamental for its oxidative and nutritional quality. For this purpose, agronomic and technological conditions of its production have been investigated. It has also been examined how this fraction can be better preserved during storage. Moreover, its relation with VOO sensorial characteristics and its interaction with a protein in emulsion foods have also been studied. Finally, an experimental work was carried out to determine the antioxidative and heat resistance properties of a new antioxidant (EVS-OL) when used for high temperature frying such as is typically employed for the preparation of french fries. Results of the scientific research have been submitted for a publication and some data has already been published in national and international scientific journals.

This PhD thesis has been proposed to validate and then apply innovative analytical methodologies for the determination of compounds with harmful impact on human health, such as biogenic amines and ochratoxin A in wines. Therefore, the influence of production technology (pH, amino acids precursor and use of different malolactic starters) on biogenic amines content in wines was evaluated. An HPLC method for simultaneous determination of amino acids and amines with precolumnderivatization with 9-Fluorenyl-methoxycarbonyl chloride (FMOC-Cl) and UV detection was developed. Initially, the influence of pH, time of derivatization, gradient profile were studied. In order to improve the separation of amino acids and amines and reduce the time of analysis, it was decided to study the influence of different flows and the use of different columns in the chromatographic method. Firstly, a C18 Luna column was used and later two monolithic columns Chromolith in series. It appeared to be suitable for an easy, precise and accurate determination of a relatively large number of amino acids and amines in wines. This method was then applied on different wines produced in the Emilia Romagna region. The investigation permitted to discriminate between red and white wines. Amino acids content is related to the winemaking process. Biogenic amines content in these wines does not represent a possible toxicological problem for human health. The results of the study of influence of technologies and wine composition demonstrated that pH of wines and amino acids content are the most important factors. Particularly wines with pH > 3,5 show higher concentration of biogenic amines than wines with lower pH. The enrichment of wines by nutrients also influences the content of some biogenic amines that are higher in wines added with amino acids precursors. In this study, amino acids and biogenic amines are not statistically affected by strain of lactic acid bacteria inoculated as a starter for malolactic fermentation. An evaluation of different clean-up (SPE-MycoSep; IACs and LLE) and determination methods (HPLC and ELISA) of ochratoxin A was carried out. The results obtained proved that the SPE clean-up are reliable at the same level while the LLE procedures shows lowest recovery. The ELISA method gave a lower determination and a low reproducibility than HPLC method.

The aim of the first part of this thesis was to evaluate the effect of trans fatty acid- (TFA), contaminant, polycyclic aromatic hydrocarbon (PAH)- and oxidation productenriched diets on the content of TFA and conjugated linoleic acid (CLA) isomers in meat and liver of both poultry and rabbit. The enriched feedings were prepared with preselected fatty co-and by-products that contained low and high levels of TFA (low, palm fatty acid distillate; high, hydrogenated palm fatty acid distillate), environmental contaminants (dioxins and PCBs) (two different fish oils), PAH (olive oil acid oils and pomace olive oil from chemical refining, for low and high levels) and oxidation products (sunflower-olive oil blend before and after frying), so as to obtain single feedings with three enrichment degrees (high, medium and low) of the compound of interest. This experimental set-up is a part of a large, collaborative European project (http://www.ub.edu/feedfat/), where other chemical and health parameters are assessed. Lipids were extracted, methylated with diazomethane, then transmethylated with 2N KOH/methanol and analyzed by GC and silver-ion TLC-GC. TFA and CLA were determined in the fats, the feedings, meat and liver of both poultry and rabbit. In general, the level of TFA and CLA in meat and liver mainly varied according to those originally found in the feeding fats. It must be pointed out, though, that TFA and CLA accumulation was different for the two animal species, as well as for the two types of tissues. The TFA composition of meat and liver changes according to the composition of the oils added to the feeds with some differences between species. Chicken meat with skin shows higher TFA content (2.6–5.4 fold) than rabbit meat, except for the “PAH” trial. Chicken liver shows higher TFA content (1.2–2.1 fold) than rabbit liver, except for the “TRANS” and “PAH” trials. In both chicken and rabbit meats, the TFA content was higher for the “TRANS” trial, followed by the “DIOXIN” trial. Slight differences were found on the “OXIDATION” and “PAH” trends in both types of meats. In both chicken and rabbit livers, the TFA content was higher for the “TRANS” trial, followed by those of the “PAH”, “DIOXIN” and “OXIDATION” trials. This trend, however, was not identical to that of feeds, where the TFA content varied as follows: “TRANS” > “DIOXIN” >“PAH” > “OXIDATION”. In chicken and rabbit meat samples, C18:1 TFA were the most abundant, followed by C18:2 TFA and C18:3 TFA, except for the “DIOXIN” trial where C18:3 TFA > C18:2 TFA. In chicken and rabbit liver samples of the “TRANS” and “OXIDATION” trials, C18:1 TFA were the most abundant, followed by C18:2 TFA and C18:3 TFA, whereas C18:3 TFA > C18:2 in the “DIOXIN” trial. Slight differences were found on the “PAH” trend in livers from both species. The second part of the thesis dealt with the study of lipid oxidation in washed turkey muscle added with different antioxidants. The evaluation on the oxidative stability of muscle foods found that oxidation could be measured by headspace solid phase microestraction (SPME) of hexanal and propanal. To make this method effective, an antioxidant system was added to stored muscle to stop the oxidative processes. An increase in ionic strength of the sample was also implemented to increase the concentration of aldehydes in the headspace. This method was found to be more sensitive than the commonly used thiobarbituric acid reactive substances (TBARs) method. However, after antioxidants were added and oxidation was stopped, the concentration of aldehydes decreased. It was found that the decrease in aldehyde concentration was due to the binding of the aldehydes to muscle proteins, thus decreasing the volatility and making them less detectable.

Maintaining the postharvest quality of whole and fresh-cut fruit during storage and distribution is the major challenge facing fruit industry. For this purpose, industry adopt a wide range of technologies to enable extended shelf-life. Many factors can lead to loss of quality in fresh product, hence the common description of these products as ‘perishable’. As a consequence normal factors such as transpiration and respiration lead ultimately to water loss and senescence of the product. Fruits and vegetables are living commodities and their rate of respiration is of key importance to maintenance of quality. It has been commonly observed that the greater the respiration rate of a product, the shorter the shelf-life. The principal problem for fresh-cut fruit industries is the relative shorter shelf-life of minimally processed fruit (MPF) compared to intact product. This fact is strictly connected with the higher ethylene production of fruit tissue stimulated during fresh-cut processing (peeling, cutting, dipping). 1-Methylcyclopropene (1-MCP) is an inhibitor of ethylene action and several researches have shown its effectiveness on the inhibition of ripening and senescence incidence for intact fruit and consequently on their shelf-life extension. More recently 1-MCP treatment has been tested also for shelf-life extension of MPF but discordant results have been obtained. Considering that in some countries 1-MCP is already a commercial product registered for the use on a number of horticultural products, the main aim of this actual study was to enhance our understanding on the effects of 1-MCP treatment on the quality maintenance of whole and fresh-cut climacteric and non-climacteric fruit (apple, kiwifruit and pineapple). Concerning the effects of 1-MCP on whole fruit, was investigated the effects of a semi-commercial postharvest treatment with 1-MCP on the quality of Pink Lady apples as functions of fruit ripening stage, 1-MCP dose, storage time and also in combination with controlled atmospheres storage in order to better understand what is the relationship among these parameters and if is possible to maximize the 1-MCP treatment to meet the market/consumer needs and then in order to put in the market excellent fruit. To achieve this purpose an incomplete three-level three-factor design was adopted. During the storage were monitored several quality parameters: firmness, ripening index, ethylene and carbon dioxide production and were also performed a sensory evaluations after 6 month of storage. In this study the higher retention of firmness (at the end of storage) was achieved by applying the greatest 1-MCP concentration to fruits with the lowest maturity stage. This finding means that in these semi-commercial conditions we may considerate completely blocked the fruit softening. 1-MCP was able to delay also the ethylene and CO2 production and the maturity parameters (soluble solids content and total acidity). Only in some cases 1-MCP generate a synergistic effect with the CA storage. The results of sensory analyses indicated that, the 1-MCP treatment did not affect the sweetness and whole fruit flavour while had a little effect on the decreasing cut fruit flavour. On the contrary the treated apple was more sour, crisp, firm and juicy. The effects of some treatment (dipping and MAP) on the nutrient stability were also investigated showing that in this case study the adopted treatments did not have drastic effects on the antioxidant compounds on the contrary the dipping may enhance the total antioxidant activity by the accumulation of ascorbic acid on the apple cut surface. Results concerning the effects of 1-MCP in combination with MAP on the quality parameters behaviour of the kiwifruit were not always consistent and clear: in terms of colour maintenance, it seemed to have a synergistic effect with N2O MAP; as far as ripening index is concerned, 1-MCP had a preservative effect, but just for sample packed in air.

No description available.

The aim of the present study was to examine the association between milk protein polymorphism and fatty acids profiles of bovine milk. Milk samples were collected from each of 55 Reggiana cows during early, mid and late lactation, respectively, in two farms within the production area of Parmigiano Reggiano cheese. Identification and quantification of fatty acids were performed by gas chromatography. Milk fatty acid composition using cows of differing κ-casein (κ-Cn) and β-lactoglobulin (β-Lg) phenotypes was investigated. Statistically significant results regarding the associations between milk fatty acid composition and κ-Cn phenotype were found, in particular, κ-Cn BB seems to influence de novo fatty acid synthesis in the mammary gland. Also κ-Cn AB seems to have the same effect. Proportions of C10:0 (2,29a AA; 2,53b AB; 2,59b BB), C12:0 (2,77a AA; 3,17b AB; 3,20b BB) and C14:0 (9,22a AA; 10,25b AB; 10,27b BB) were higher in the milk from cows with κ-Cn phenotype AB and BB vs κ-Cn phenotype AA (p<0,05). Conversely C18:0 (7,84b AA; 7,20a,b AB; 6,94a BB) and C18:1 (19,19b AA; 16,81a AB; 16,79a BB) were lower in the milk from cows with κ-Cn phenotype AB and BB vs κ-Cn phenotype AA. The association between milk fatty acid composition and β-Lg phenotype was not statistically significant, except for some fatty acids. In particular, C12:0 (3,05a AA; 3,04a AB; 3,33b BB) was higher in the milk from cows with β-Lg phenotype BB vs β-Lg phenotype AA and AB (p<0,05). Concentrations of C18:0 (6,93a AA; 7,86b AB; 6,59a BB) and C18:1 (16,74a,b AA; 18,24b AB; 16,07a BB) were lower in the milk from cows with β-Lg phenotype AA and BB vs β-Lg phenotype AB (p<0,05). Moreover this research, carried out in farms within the Parmigiano Reggiano cheese district, analysed also the size distribution of fat globules in bulk milk of Reggiana and Frisona breed cows. In particular, the size distribution of individual milk fat globules of Reggiana cows with differing κ-Cn phenotypes was considered. From first observations, no statistically significant differences were observed.

"Bioactive compounds" are extranutritional constituents that typically occur in small quantities in food. They are being intensively studied to evaluate their effects on health. Bioactive compounds include both water soluble compounds, such as phenolics, and lipidic substances such as n-3 fatty acids, tocopherols and sterols. Phenolic compounds, tocopherols and sterols are present in all plants and have been studied extensively in cereals, nuts and oil. n-3 fatty acids are present in fish and all around the vegetable kingdom. The aim of the present work was the determination of bioactive and potentially toxic compounds in cereal based foods and nuts. The first section of this study was focused on the determination of bioactive compounds in cereals. Because of that the different forms of phytosterols were investigated in hexaploid and tetraploid wheats. Hexaploid cultivars were the best source of esterified sterols (40.7% and 37.3% of total sterols for Triticum aestivum and Triticum spelta, respectively). Significant amounts of free sterols (65.5% and 60.7% of total sterols for Triticum durum and Triticum dicoccon, respectively) were found in the tetraploid cultivars. Then, free and bound phenolic compounds were identified in barley flours. HPLCESI/ MSD analysis in negative and positive ion mode established that barley free flavan-3- ols and proanthocyanidins were four dimers and four trimers having (epi)catechin and/or (epi)gallocatechin (C and/or GC) subunits. Hydroxycinnamic acids and their derivatives were the main bound phenols in barley flours. The results obtained demonstrated that barley flours were rich in phenolic compounds that showed high antioxidant activity. The study also examined the relationships between phenolic compounds and lipid oxidation of bakery. To this purpose, the investigated barley flours were used in the bakery production. The formulated oven products presented an interesting content of phenolic compounds, but they were not able to contain the lipid oxidation. Furthermore, the influence of conventional packaging on lipid oxidation of pasta was evaluated in n-3 enriched spaghetti and egg spaghetti. The results proved that conventional packaging was not appropriated to preserve pasta from lipid oxidation; in fact, pasta that was exposed to light showed a high content of potentially toxic compounds derived from lipid oxidation (such as peroxide, oxidized fatty acids and COPs). In the second section, the content of sterols, phenolic compounds, n-3 fatty acids and tocopherols in walnuts were reported. Rapid analytical techniques were used to analyze the lipid fraction and to characterize phenolic compounds in walnuts. Total lipid chromatogram was used for the simultaneous determination of the profile of sterols and tocopherols. Linoleic and linolenic acids were the most representative n-6 and n-3 essential dietary fatty acids present in these nuts. Walnuts contained substantial amounts of γ- and δ-tocopherol, which explained their antioxidant properties. Sitosterol, Δ5-avenasterol and campesterol were the major free sterols found. Capillary electrophoresis coupled to DAD and microTOF was utilized to determine phenolic content of walnut. A new compound in walnut ((2E,4E)- 8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-β-D-glucopiranosyl ester, [M−H]− 403.161m/z) with a structure similar to glansreginins was also identified. Phenolic compounds corresponded to 14–28% of total polar compounds quantified. Aglycone and glycosylated ellagic acid represented 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-β-D-glucopiranosyl ester was in the range of 72–86% of total quantified compounds.

This PhD thesis describes the application of some instrumental analytical techniques suitable to the study of fundamental food products for the human diet, such as: extra virgin olive oil and dairy products. These products, widely spread in the market and with high nutritional values, are increasingly recognized healthy properties although their lipid fraction might contain some unfavorable components to the human health. The research activity has been structured in the following investigations: “Comparison of different techniques for trans fatty acids analysis” “Fatty acids analysis of outcrop milk cream samples, with particular emphasis on the content of Conjugated Linoleic Acid (CLA) and trans Fatty Acids (TFA), by using 100m high-polarity capillary column” “Evaluation of the oxidited fatty acids (OFA) content during the Parmigiano-Reggiano cheese seasoning” “Direct analysis of 4-desmethyl sterols and two dihydroxy triterpenes in saponified vegetal oils (olive oil and others) using liquid chromatography-mass spectrometry” “Quantitation of long chain poly-unsatured fatty acids (LC-PUFA) in base infant formulas by Gas Chromatography, and evaluation of the blending phases accuracy during their preparation” “Fatty acids composition of Parmigiano Reggiano cheese samples, with emphasis on trans isomers (TFA)”