Tracciabilit\ue0 dell'origine del latte alpino mediante lo studio del profilo alcaloidico naturale

In 2015, 162.8 million tonnes of cow\u2019s milk were produced in the EU-28, headed by Germany and France with 33 and 26 million tonnes respectively, while Italy was in seventh place with 11 million tonnes (Eurostat statistics explained), and world milk production is forecast to grow. In a globalised market, demand is growing for secure information about product traceability, and the consumer trend is increasingly moving towards conscientious purchasing, which avoids adulteration and fraud. Traceability is a powerful tool for pursuing different objectives: reinforcing consumers\u2019 trust and loyalty to a product, peace of mind to promote product preference, and increasing confidence in the overall food chain quality system. As regards milk, in Italy the Decreto Interministeriale of 9 December 2016 made indication of the origin of raw milk materials on the packaging label mandatory, as stated by Regulation (EU) No. 1169/2011. In the same way, not only is the country of origin important, but also the differentiation between milk from highland and lowland areas, due to the evident implications for food quality. The different dietary regime of animals is the basis of the characteristics distinguishing alpine milk from intensive farming milk, and analytical controls usually exploit these properties. This work aimed to propose new markers for alpine dairy product traceability by focusing on one of the most abundant and ubiquitous groups of secondary metabolites in plants, namely alkaloids. In this work we developed a combined targeted and untargeted screening method for alkaloid profiling, using liquid chromatography coupled with high resolution mass spectrometry. Online SPE pre-treatment of herbal and milk sample extracts was proposed in order to reduce the impact of the matrix effect on instrumental response. Quantification of 41 analytes with reference to pure analytical standards, and putative identification of a further 116 alkaloids, confirmed on the basis of accurate mass, isotopic pattern, chromatographic retention time and fragmentation profile, were proposed. Moreover, the alkaloid profiles of over 60 alpine herbs sampled in two natural pastures in the eastern Italian Alps were defined, providing evidence that alkaloid composition represents an interesting tool for individually characterising plant families, with the most encouraging results for the Poaceae species. The composition and variability of alkaloids ingested by dairy cows grazing on the two grasslands was also investigated, verifying the possibility of discriminating animal diets from different pastures. Lastly, the variability of alkaloid profiles for milk samples produced by cows grazing on the two pastures was defined and the possibility of discriminating them with regard to pasture origin

The controversial relationship between chitosan and the microorganisms involved in the production of fermented beverages

Chitosan is a promising antimicrobial agent available in the beverage industry, because it ensures the control of a wide range of spoilage microorganisms. As chitosan does not alter the characteristics of fermented beverages, it is nowadays widely employed in the wine sector. In this work, an exhaustive chemical characterization of 12 commercial chitosans was performed in accordance with the OIV methods. These analyses made it possible to confirm or determine the animal or fungal origin of the 12 samples. Furthermore, ionic chromatography coupled with an amperometric detector (IC-PAD) confirmed peculiar polysaccharide profiles for fungal and animal-derived chitosans. The antimicrobial activity of chitosans was evaluated against the microorganisms involved in beverage fermentation or capable spoil wine, beer and cider. Chitosans were tested in static and stirred conditions, in a synthetic medium that reproduces fermented beverage conditions, to discriminate against the physical settling of cells and their specific antimicrobial activity. Moreover, the activity of the soluble portion of chitosan was checked by inoculating microorganisms in the media after chitosans removal. The results highlighted the different sensitivity of microorganisms to chitosans, allowing selective control of spoilage agents. However, the yeast and bacteria involved in fermentation were damaged by chitosan, and the synthetic media treated with this molecule showed a less fermentative aptitude. These results suggest that chitosan is a promising tool in fermented beverage production, but an in-depth study of the biochemical interaction between chitosan and food microorganisms is necessar

Atypical ageing defect in Pinot Blanc wines: a comparison between organic and conventional production management systems

Atypical ageing (ATA) is an aroma defect that occurs in white wines and entails a loss of varietal aromas as well as scents of wet mop, shoe polish and dish rag. 2-Aminoacetophenone (2AAP) - a degradation product of indole-3-acetic acid (IAA) - has been described as the main odour-active compound and chemical marker responsible for this off-flavour. A stress reaction in the vineyard triggered by climatic, pedological and viticultural factors can ultimately cause ATA development in wines and remarkably affect wine quality. The aim of this research was to investigate the influence of three grapevine management systems on the occurrence of ATA. The experiments were carried out on Pinot Blanc grape samples from vines cultivated using one conventional and two organic approaches. The management systems mainly differed for the fertilisation regime and the weed control

Evaluation of antioxidant supplementation in must on the development and potential reduction of different compounds involved in atypical ageing of wine using HPLC-HRMS

Atypical aging of white wines (ATA) is an off-flavour characterised by rapid loss of fruity aromas and the development of unpleasant odours. This study aims to evaluate the effectiveness of different oenological adjuvants (4-ethylcatechol, 4-methylcatechol, gentisic acid, hydroxytyrosol, ascorbic acid, glutathione, ellagic acid, gallic acid, galla tannin, ellagic tannin, quebracho, grape tannins) added to musts before fermentation for preventing the possible development of ATA. High resolution mass spectrometry made it possible to quantify and qualify free and conjugated IAA in wine at the end of the fermentation and subsequently 2-aminoacetophenone (AAP) was quantified after the force ageing period (6 days at 40 °C). Ascorbic acid was confirmed as the most appropriate antioxidant adjuvant which can be used for ATA defect prevention. With an almost comparable effect, gallotannin addition prevented AAP production to exceed 1 μg/L. A predicted model (ANCOVA) indicated that over 80% of the variability of potential AAP formation in wines was explained by the amount of precursors, grape variety and antioxidant treatment. Moreover, a suspect screening approach made it possible to study the kinetic formation and the consumption of the reaction metabolites formed during the oxidative degradation of IAA leading to AA

In Vitro bioaccessibility of bioactive compounds from Citrus pomaces and orange pomace biscuits

The present investigation aimed to provide novel information on the chemical composition and in vitro bioaccessibility of bioactive compounds from raw citrus pomaces (mandarin varieties Clemenule and Ortanique and orange varieties Navel and Valencia). The effects of the baking process on their bioaccessibility was also assessed. Samples of pomaces and biscuits containing them as an ingredient were digested, mimicking the human enzymatic oral gastrointestinal digestion process, and the composition of the digests were analyzed. UHPLC-MS/MS results of the citrus pomaces flavonoid composition showed nobiletin, hesperidin/neohesperidin, tangeretin, heptamethoxyflavone, tetramethylscutellarein, and naringin/narirutin. The analysis of the digests indicated the bioaccessibility of compounds possessing antioxidant [6.6–11.0 mg GAE/g digest, 65.5–97.1 µmol Trolox Equivalents (TE)/g digest, and 135.5–214.8 µmol TE/g digest for total phenol content (TPC), ABTS, and ORAC-FL methods, respectively; significant reduction (p < 0.05) in Reactive Oxygen Species (ROS) formation under tert-butyl hydroperoxide (1 mM)-induced conditions in IEC-6 and CCD-18Co cells when pre-treated with concentrations 5–25 µg/mL of the digests], anti-inflammatory [significant reduction (p < 0.05) in nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophages], and antidiabetic (IC50 3.97–11.42 mg/mL and 58.04–105.68 mg/mL for α-glucosidase and α-amylase inhibition capacities) properties in the citrus pomaces under study. In addition, orange pomace biscuits with the nutrition claims “no-added sugars” and “source of fiber”, as well as those with good sensory quality (6.9–6.7, scale 1–9) and potential health promoting properties, were obtained. In conclusion, the results supported the feasibility of citrus pomace as a natural sustainable source of health-promoting compounds such as flavonoids. Unfractionated orange pomace may be employed as a functional food ingredient for reducing the risk of pathophysiological processes linked to oxidative stress, inflammation, and carbohydrate metabolism, such as diabetes, among other

Connexin Hemichannel Activation by S-Nitrosoglutathione Synergizes Strongly with Photodynamic Therapy Potentiating Anti-Tumor Bystander Killing

In this study, we used B16-F10 cells grown in the dorsal skinfold chamber (DSC) preparation that allowed us to gain optical access to the processes triggered by photodynamic therapy (PDT). Partial irradiation of a photosensitized melanoma triggered cell death in non-irradiated tumor cells. Multiphoton intravital microscopy with genetically encoded fluorescence indicators revealed that bystander cell death was mediated by paracrine signaling due to adenosine triphosphate (ATP) release from connexin (Cx) hemichannels (HCs). Intercellular calcium (Ca2+) waves propagated from irradiated to bystander cells promoting intracellular Ca2+ transfer from the endoplasmic reticulum (ER) to mitochondria and rapid activation of apoptotic pathways. Combination treatment with S-nitrosoglutathione (GSNO), an endogenous nitric oxide (NO) donor that biases HCs towards the open state, greatly potentiated anti-tumor bystander killing via enhanced Ca2+ signaling, leading to a significant reduction of post-irradiation tumor mass. Our results demonstrate that HCs can be exploited to dramatically increase cytotoxic bystander effects and reveal a previously unappreciated role for HCs in tumor eradication promoted by PDT

Verifying the botanical authenticity of commercial tannins through sugars and simple phenols profiles

Commercial tannins from several botanical sources and with different chemical and technological characteristics are used in the food and winemaking industries. Different ways to check their botanical authenticity have been studied in the last few years, through investigation of different analytical parameters. This work proposes a new, effective approach based on the quantification of 6 carbohydrates, 7 polyalcohols, and 55 phenols. 87 tannins from 12 different botanical sources were analysed following a very simple sample preparation procedure. Using Forward Stepwise Discriminant Analysis, 3 statistical models were created based on sugars content, phenols concentration and combination of the two classes of compounds for the 8 most abundant categories (i.e. oak, grape seed, grape skin, gall, chestnut, quebracho, tea and acacia). The last approach provided good results in attributing tannins to the correct botanical origin. Validation, repeated 3 times on subsets of 10% of samples, confirmed the reliability of this mode