Traditional decoction and PUAE aqueous extracts of pomegranate peels as potential low-cost anti-tyrosinase ingredients

open6The aim of the study is to evaluate the anti-tyrosinase activity of dierent aqueous extracts obtained from pomegranate juice processing by-products. External pomegranate peels of two certified cultivars (Akko and Wonderful), were extracted using only water as the extraction solvent. A traditional decoction and a pulsed ultrasound-assisted extraction (PUAE), both 10 min long, were performed and compared. All the aqueous extracts proved to be rich in bioactive compounds. In particular, the total phenolic content (TPC) ranged from 148 to 237 mg gallic acid equivalent (GAE)/g of dried peels (DW), the radical-scavenging ability (RSA) ranged from 307 to 472 mg ascorbic acid equivalent (AAE)/g DW, the free ellagic acid content (EA) ranged from 49 to 94 g/mL, and the ellagitannins (ETs) ranged from 242 to 340 g/mL. For both cultivars, PUAE extracts had higher ET content and a lower EC50, while the decoctions had slightly higher TPC, RSA, and free EA amounts. Principal component analysis (PCA) highlighted the direct correlation between the ET content and the tyrosinase enzyme inhibition (lower values of EC50). These findings suggest the potential use of both these natural extracts as low-cost lightening and/or anti-browning ingredients exploitable in several formulations (e.g., cosmetics) or extemporarily usable.openFEDERICA TURRINI; PAOLA MALASPINA; PAOLO GIORDANI; SILVIA CATENA; PAOLA ZUNIN; RAFFAELLA BOGGIA;Turrini, Federica; Malaspina, Paola; Giordani, Paolo; Catena, Silvia; Zunin, Paola; Boggia, Raffaell

Microdispersions of ellagic acid and pomegranate extracts as new potential nutraceutical ingredients

The health properties attributed to several fruits (i.e. pomegranates, raspberries, strawberries, blackberry, chestnuts, walnuts, pecan), herbs (tea) and seeds (berries seeds) are attributed to an important group of natural polyphenols classified as hydrolysable tannins (HT) named Ellagitannins (ETs), that have shown in vitro multi-target biological properties relevant to the treatment of several human diseases. In vivo, ETs are rather not absorbed, and they are hydrolysed providing mainly Ellagic acid (EA). EA is endowed with the same biological properties of ETs and it could be considered as the responsible of their health benefits. Unfortunately, EA cannot be exploited for in vivo applications because of its poor water solubility (9.7 \u3bcg/mL) and accordingly low bioavailability. At first, aiming to increase EA solubility, an EA solid microdispersion (EA-md) was realized by employing only water and low methoxylated pectin, as a food compatible excipient, by applying spray drying technology. EA-md showed a 22% (w/w) Drug Loading (DL), a 30 times improved water solubility maintaining a remarkable radical scavenging activity [1]. It has been analytically characterised and used for in vivo pharmacological treatments in order to evaluate it as potential nutraceutical ingredient. Adult (3-6 months old) and old (20-22 old months) male mice were chronically administered EA-md dissolved in the drinking water (about 150 mg / Kg) for 14 days. During this period, animals were monitored for the spontaneous motor activity and for curiosity before, during and at the end of the EA-md treatment. Adult and old mice were then sacrificed for \u201cex vivo, in vitro\u201d analysis to test the efficiency of noradrenaline release from cortical nerve endings. It is known that noradrenaline exocytosis from cortical nerve endings is significantly impaired during ageing. We found that the chronic administration of EA-md did not alter the noradrenaline exocytosis from cortical nerve endings of adult mice, but significantly recovered the reduced noradrenaline overflow in aged mice. Further investigations are needed to explore the cellular cascade of events accounting for the beneficial effect. In a second step, pomegranate, as a natural source of EA, has been considered to similarly prepare and investigate an analogous formulation. Since pomegranate fruit is recognized as one of the most important sources of ETs, mainly localized in the by-products obtained after industrial juice squeezing, a method to convert the squeezing marcs into a potential nutraceutical ingredient has been explored. In particular, Pulsed Ultrasound-Assisted Extraction (PUAE), using just water as solvent, resulted to be suitable for extracting the water-soluble bioactive molecules (PEx), whose content in hydrolysable tannins, standardized in EA, has been determined. Furthermore, the already mentioned spray drying microdispersion has been employed to formulate and to stabilize it over time. This last formulation (PEx-md) will be subjected to the already mentioned pharmacological experiments in order to study its nutraceutical properties too. [1] S. Alfei, F. Turrini, S. Catena, P. Zunin, B. Parodi, G. Zuccari, A.M. Pittaluga, R. Boggia, New J. Chem, 43, 2438-2448 DOI: 10.1039/C8NJ05657

Neuroinflammation in Aged Brain: Impact of the Oral Administration of Ellagic Acid Microdispersion

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 "ex-vivo, in vitro" 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

the 3rs reduction and refinement through a multivariate statistical analysis approach in a behavioural study to unveil anxiolytic effects of natural extracts of tilia tomentosa

We propose a multivariate statistical approach based on Principal Component Analysis (PCA) as an useful instrument to improve the Rules of Refinement and Reduction in in vivo animal experimentation. We analysed with PCA the preliminary data from a study on the effects of the oral administration of Tilia tomentosa bud extracts (TTBEs) on the behavioural skills of adult and aged male and female mice. PCA allows to rationalize the data set information and to dissect the results, showing connections among variables under study (behavioural parameters) and different trends in the experimental groups (control and TTBEs-administered animals). Our results show that PCA can give some important information that can be useful for the refinement of the experimental protocol, in order to reduce the number of the animals used in the experiments and/or the behavioural tests to get reliable information

Determination of free amino acids in infant formulas.

none2A 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 signi"cant amounts of free arginine, tyrosine and methionine were detected, although they had not been listed among the added ingredientsmixedP. Zunin; F. EvangelistiZunin, Paola; Evangelisti, Filipp

Valutazione dell\u2019autenticit\ue0 dei succhi di frutta mediante fingerprint spettrofotometrica UV_visibile

Le recenti Direttive Europee, recepite quest\u2019anno anche in Italia, concernenti i succhi di frutta e altri prodotti analoghi hanno reso obbligatorio, da parte delle industrie alimentari, la dichiarazione in etichetta del tipo di frutta caratterizzante il prodotto che dovr\ue0 essere indicato nella denominazione commerciale del succo [1]. Tale normativa ha come fine ultimo quello di impedire che, ad esempio, un prodotto composto dal 70% di succo di mela e dal 30% di succo di melagrane venga etichettato come \u201csucco di melagrane\u201d, ingannando il consumatore. Di conseguenza, la presenza e lo sviluppo di metodi di analisi per assicurare l'autenticit\ue0 dei succhi di frutta, e quindi di ci\uf2 che viene dichiarato in etichetta, \ue8 di interesse comune sia per il settore industriale al fine di salvaguardare prodotti di qualit\ue0, sia per i consumatori. Se da un lato, la sostituzione anche parziale di un frutto con un altro, rappresenta un danno economico, dall\u2019altro l\u2019omissione di alcuni ingredienti, presenti, ma non dichiarati, potrebbe avere ripercussioni anche gravi sulla salute dei consumatori (es. allergeni nascosti). La maggior parte delle metodiche analitiche, ad oggi riportate in letteratura per l\u2019autenticazione dei succhi di frutta, implicano l\u2019utilizzo di strumenti ad alto costo, l\u2019impiego sequenziale di numerosi strumenti o l\u2019uso di grandi quantit\ue0 di solventi. Inoltre, tali metodiche sono spesso onerose sia da un punto di vista dei costi che dei tempi di analisi [2]. Per ovviare a tali inconvenienti, nel laboratorio di Chimica degli Alimenti dell\u2019Universit\ue0 di Genova \ue8 da qualche tempo in studio un\u2019alternativa analitica rapida, efficace ed economica, da utilizzare quanto meno in una fase di screening, al fine di evidenziare rapidamente alcune delle pi\uf9 tipiche azioni illecite operate in questo settore [3]. Il metodo analitico proposto si basa sulla spettroscopia UV-visibile e sull\u2019analisi multivariata dei dati. La spettrofotometria UV-visibile, una metodica analitica classica diffusa in quasi tutti i laboratori e notoriamente poco costosa e di facile impiego, \ue8 stata per\uf2 utilizzata non in maniera tradizionale, vale a dire valutando l\u2019assorbimento a singole determinate lunghezze d\u2019onda, bens\uec come tecnica aspecifica in cui per ogni campione analizzato \ue8 stato valutato l\u2019intero spettro UV-visibile (190\u20131100 nm). Pertanto ad ogni campione analizzato corrisponde uno spettro completo, che pu\uf2 essere considerato come una sorta d\u2019impronta digitale (fingerprint) del campione stesso. In questo modo \ue8 stata valutata la possibilit\ue0 di evidenziare andamenti spettrali differenti a seconda della diversa origine dei succhi presi in esame. In particolare sono stati studiati sia succhi preparati in laboratorio a partire da frutta fresca (considerati \u201cautentici\u201d), sia succhi commerciali. Sono state successivamente preparate miscele binarie e ternarie di succhi diversi, mediante l\u2019ausilio di tecniche di mixture experimental design, al fine di testare la metodica analitica su campioni appositamente adulterati. Infine, i risultati ottenuti sono stati confrontati con quelli provenienti da database compositivi

Oxidative stability of virgin olive oil enriched with carnosic acid

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

\u2018Specialty\u2019 or \u2018Gourmet\u2019 oils: a multivariate statistical approach for the rapid identification of their botanical species

In the last decade, the increase of consumers\u2019 preferences for the pleasant sensorial features of oils of different botanical origin has favored the diffusion of the so-called \u2018specialty\u2019 or \u2018gourmet\u2019 oils, namely cold-pressed industrial virgin oils obtained from seeds, fruits, and nuts1. Besides their flavor and aroma, the peculiar features of \u2018gourmet\u2019 oils lay in their alleged health promoting activities2, and in their low global production amounts, which result in their high costs. However, the recent market deregulation of virgin and/or cold pressed vegetable oils, and at the same time their high commercial value, asks for analytical methods which assess their authenticity. The aim of this study was evaluating the possibility to identify the botanical origin of the oil-based raw material by the routine analysis of their fatty acid composition coupled with chemometric methods and to explore also a new untargeted approach to their identification applying just their color analysis. A comprehensive data collection of authentic \u2018gourmet\u2019 oils, belonging to eight different botanical species (i.e., Almond, Apricot, Avocado, Hazelnut, Mosqueta rose, Rosehip, Sunflower, and Walnut) plus Olive oil as the gold standard of cold-pressed virgin oils, was performed. Their fatty acid composition has been re-elaborated in a statistical multivariate approach applying Principal Component Analysis (PCA) and classification methods. Moreover, a simple, rapid, and non-destructive spectrophotometric analysis has been considered to discriminate among different species. Particularly, the raw diffuse reflectance spectra recorded in the spectral range 380-780 nm by a UV-Vis spectrophotometer endowed with an integrating sphere was taken into account and elaborated by chemometrics. This information was compared with the results obtained by the most common approach based on the CIELab parameters and a fusion matrix of chromatographic and spectral data was also investigated. Both the fatty acid composition and the color (total spectrum and colorimetric parameters) of these vegetal oils have proven to be two promising markers of their botanical authenticity. [1] N. Cicero, A. Albergamo, A. Salvo, G.D. Bua, G. Bartolomeo, V. Mangano, A. Rotondo, V. Di Stefano, G. Di Bella, G. Dugo, Food Res. Int. 2018, 109, 517\u2013525. [2] E.M. Hernandez, Functional Dietary Lipids\u2014Food Formulation. 2016, 69\u2013101

Sterol oxidation in infant milk formulas and milk cereals.

none37-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.mixedP. Zunin; C. Calcagno; F. EvangelistiZunin, Paola; Calcagno, Claudio; Evangelisti, Filipp