From Field to Waste Valorization: A Preliminary Study Exploring the Impact of the Wine Supply Chain on the Phenolic Profile of Three Sardinian Pomace Extracts.

The winemaking process generates an annual global production of about 10 million tons of waste consisting of stalks, skin, and seeds. The possible reutilization of wine pomace is strictly linked to its chemical composition. In this preliminary study, three different Sardinian white grapes (Malvasia, Vermentino and Nasco) grown in the same area were evaluated through a whole wine production chain. To reduce environmental impact, all the grapes were treated following the integrated production practice (IPP) strategies. The adopted agronomic methods and the main physico-chemical parameters of the fresh fruits and musts were evaluated. A fully qualitative and quantitative characterization of the phenolic fraction of the pomace extracts was performed by HPLC-DAD after a post-winemaking process. Water and ethanol were utilized as green solvents in the extraction process. Additionally, the entire pomace post-winemaking process was carried out within the winery facilities to reduce energy loss and road transportation. The findings demonstrated that large amounts of beneficial polyphenols are present in pomace extracts, and that the type of grape used, agronomic practices, and winemaking method all influence the quantity and quality of the extracts. The polyphenol concentrations in the Vermentino (28,391.5 ± 7.0 mg/kg) and Malvasia pomace (11,316.3 ± 6.5 mg/kg) were found to be the highest and lowest, respectively.

Determination of Pesticide Residues in IV Range Artichoke (Cynara cardunculus L.) and Its Industrial Wastes.

Fourth-range products are those types of fresh fruit and vegetables that are ready for raw consumption or after cooking, and belong to organic or integrated cultivations. These products are subject to mild post-harvesting processing procedures (selection, sorting, husking, cutting, and washing), and are afterwards packaged in packets or closed food plates, with an average shelf life of 5-10 days. Artichokes are stripped of the leaves, stems and outer bracts, and the remaining heads are washed with acidifying solutions. The A LC-MS/MS analytical method was developed and validated following SANTE guidelines for the detection of 220 pesticides. This work evaluated the distribution of pesticide residues among the fraction of artichokes obtained during the industrial processing, and the residues of their wastes left on the field were also investigated. The results showed quantifiable residues of one herbicide (pendimethalin) and four fungicides (azoxystrobin, propyzamide, tebuconazole, and pyraclostrobin). Pendimethalin was found in all samples, with the higher values in leaves 0.046 ± 8.2 mg/kg and in field waste 0.30 ± 6.7 mg/kg. Azoxystrobin was the most concentrated in the outer bracts (0.18 ± 2.9 mg/kg). The outer bracts showed the highest number of residues. The industrial waste showed a significant decrease in the number of residues and their concentration.

A Cocktail-Based Formula for the Design of Nanosized Cosmeceuticals as Skincare and Anti-Age Products.

Nasco and Bovale grape pomace extracts, alone or in association, were loaded in nanoemulsions tailored for cosmetic application, using Kolliphor®RH40 (kolliphor) as the synthetic surfactant, Olivem®1000 (olivem) as the natural one, and lecithin as the cosurfactant. Pink transparent or milky dispersions, as a function of the used extract and surfactant, were obtained to be used as cosmeceutical serum or milk. The sizes of the nanoemulsion droplets were small (≈77 nm with kolliphor and ≈141 nm with olivem), homogenously dispersed (~0.24 with kolliphor and ~0.16 with olivem), highly negatively charged (≈-43 mV irrespective of the used surfactant) and their stability either on storage or under stressing conditions was affected by the used extract and surfactant. Formulations protected the extracts from the degradation caused by UV exposition, were biocompatible against keratinocytes, protected them against oxidative damages induced using hydrogen peroxide and inhibited the release of nitrite induced in macrophages using the lipopolysaccharide inflammatory stimulus. The overall results underlined the key role played by the composition of the formula to achieve a suitable cosmeceutical for skin care but even for the prevention of premature aging and chronic damages caused by the stressing conditions.

Design and in vitro effectiveness evaluation of Echium amoenum extract loaded in bioadhesive phospholipid vesicles tailored for mucosal delivery.

The Echium amoenum Fisch. and C.A. Mey. (E. amoenum) is an herb native from Iranian shrub, and its blue-violet flowers are traditionally used as medical plants. In the present study, an antioxidant phytocomplex was extracted from the flowers of E. amoenum by ultrasounds-assisted hydroalcoholic maceration. The main components, contained in the extract, have been detected using HPLC-DAD, and rosmarinic acid was found to be the most abundant. The antioxidant power of the extract along with the phenolic content were measured using colorimetric assays. The extract was loaded in liposomes, which were enriched adding different bioadhesive polymers (i.e., mucin, xanthan gum and carboxymethyl cellulose sodium salt) individually or in combination. The main physico-chemical properties (i.e. size, size distribution, surface charge) of the prepared vesicles were measured as well as their stability on storage. The viscosity of dispersion and the ability of vesicles to interact with mucus were evaluated measuring their stability in a mucin dispersion and mobility in a mucin film. The biocompatibility and the ability of the formulations to protect keratinocytes from damages caused by hydrogen peroxide and to promote the cell migration were measured in vitro.

Influence of Salting Technology on the Diffusion of NaCl in Swordfish (Xiphias gladius) Fillets.

Swordfish is the most widespread billfish in the aquatic environment. The industrial processing of swordfish fillets involves salting, drying, and smoking steps. Salting techniques, dry or wet, are the most common method of fish preservation. This work evaluated salt diffusion in swordfish fillets after traditional dry salting and wet industrial injection salting methods. The data obtained from the dry salting studies highlighted that the salt diffusion process in swordfish meat was an unfavorable process depending on the contact time with the salt/meat. Moreover, irregularly shaped fillets negatively affected the salt migration in the different areas, leading to inhomogeneous and possibly unsafe final products. On the contrary, wet injection salting was suitable for processing swordfish fillets. As a result, the final products had a homogeneous salt concentration, maintained the organoleptic characteristics and health benefits for a long period, and achieved a longer shelf-life. Furthermore, the water activity (aw) values detected for the different processed fillets confirmed the physicochemical features of the final products and allow the classification of safe products. Moreover, injection salting is a quick process compatible with industrial production times.

Effect of High-Pressure Processing on Fresh Sea Urchin Gonads in Terms of Shelf Life, Chemical Composition, and Microbiological Properties.

Paracentrotus lividus is a widespread sea urchin species appreciated worldwide for the taste of its fresh gonads. High-pressure processing (HPP) can provide a thermal equivalent to pasteurization, maintaining the organoleptic properties of the raw gonads. This study evaluated HPP technology's effect at 350 MPa and 500 MPa on microbial inactivation and biochemical characteristics of P. lividus gonads. HPP at 350 MPa resulted in a higher decrease in protein and free amino acids associated with a loss of olfactory, color, and gustatory traits and a visual alteration of the texture. On the other hand, gonad samples stored for 40 days after treatments at 500 MPa showed a good organoleptic profile similar to fresh gonads. Furthermore, only 500 MPa effectively reduced mesophilic bacteria contamination among the two HPP treatments carried out. Total lipids increased during storage; however, the SAFA/PUFA rate was homogeneous during HPP trials ranging from 2.61-3.91 g/100 g. Total protein decreased more than 40% after HPP at 350 MPa, whereas, after 500 MPa, it remained stable for 20 days. The amount of free amino acid constantly decreased during storage after HPP at 350 MPa and remained constant at 500 MPa. HPP can effectively remove the bacterial flora and inactivate enzymes, maintaining the properties of the fresh sea urchin gonads.

Influence of Different Light Sources on the Biochemical Composition of Arthrospira spp. Grown in Model Systems.

Arthrospira platensis and Arthrospira maxima are prokaryotic microalgae commercially marketed as spirulina. The pigments extracted from these algae are widely used for cosmetic and nutraceutical applications. This work aimed to evaluate the influence of three light-emitting lamps (white, orange and blue) on the growth and biomass composition of two strains of A. platensis (M2 and M2M) and one of A. maxima. The obtained data show strain- and light-dependent responses of the microalgae. In addition, white and orange lights led to a similar overall effect by increasing the levels of chlorophyll a and carotenoids. However, exposure to orange light resulted in the highest dry weight (5973.3 mg L-1 in M2M), whereas white light stimulated an increase in the carbohydrate fraction (up to 42.36 g 100 g-1 in A. maxima). Conversely, blue light led to a constant increase in the concentration of phycocyanin (14 g 100 g-1 in A. maxima) and a higher content of proteins in all strains. These results provide important environmental information for modulating the growth of different spirulina strains, which can be used to address the synthesis of biochemical compounds of strategic importance for the development of new nutraceutical foods.

Effect of the Technological Process from Vine to Wine on Pesticide Residues in Vernaccia di Oristano Cultivar.

Vernaccia is a white grape mostly used to produce a distinct wine protected by the controlled designation of origin (DOC) recognition. It is very susceptible to fungal disease, and it is subjected to a defined management protocol in the field. Winemaking could influence pesticide residues through different mechanisms. This work investigated the influence on pesticide residues of the winemaking process at the industrial level of the wine Vernaccia di Oristano. Thirty-five samples of grapes, two musts, and two vines (both liquid fraction and pellets) were analyzed by using a validated multiresidue LC-MS/MS (Liquid Chromatography with tandem mass spectrometry) method. Data obtained showed the presence in grapes juice of 16 pesticides (8 not allowed in the EU) with mandipropamid and mepanipyrim, the only ones with values higher than their MRL (maximum residue level). Pesticide residues decrease in must was related to the dilution effect due to mixing the grape samples. However, pellets analysis also confirmed the high affinity of pesticides for the suspended material (fenhexamid), whereas the increase in wine to a re-solubilization process from the lees during the fermentation step. The present paper highlighted the effectiveness of the technological process of winemaking to decrease pesticide residues compared to the raw material.

Simultaneous determination and dietary risk assessment of clethodim and its metabolites in different fruits and vegetables.

Clethodim is one of the most widely used herbicides in agriculture and, after field application, is metabolised to several metabolites. The potential toxicological negative effects of these compounds are poorly understood. Thus, recently, within the risk assessment framework, the European Food Safety Authority (EFSA) proposed to include the minor metabolites in the definition of clethodim residue. In this work, an easy to use and reliable UHPLC method coupled with a triple quadrupole MS/MS was developed and validated for the detection and quantification of the herbicide clethodim and related metabolites clethodim sulphone, clethodim sulphoxide, metabolites M17R and M18R in apple, grape, olive and rice. The five analytes were extracted by using a modified QuEChERS procedure, while the active ingredients were determined in multiple reaction monitoring (MRM) ion-switching mode. The proposed method showed calibration curve linearity with r2 ≥ 0.990 for all active ingredients (a.is.) both in solvent and matrix extracts. Limits of quantitation (LOQ) of the five compounds ranged from 9.44 µg/kg for M17R in olive extract to 11.01 µg/kg for clethodim in apple extract. Recoveries values ranged from 86% to 119% at two concentration levels (LOQ and 10xLOQ), while the intraday and interday precisions of the method were both below 10% in all cases. The method was successfully used for the quantification of the five a.is. in different food matrices. Furthermore, chronic dietary risk was investigated using a hazard quotients (HQ) method based on European dietary habits. The chronic dietary exposure risk quotients ranged from 1.0 × 10-5 (lower bound scenario) to 2.7 × 10-4 (upper bound scenario) which were significantly lower than 1. Data obtained indicate that the dietary exposure risks were acceptable for clethodim and its major and minor metabolites applied in apple, table grape, rice and table olive.

Fipronil and Fipronil Sulfone Distribution in Chicken Feathers and Eggs after Oral and Dermal Exposure.

This work aimed to investigate the bio-distribution and the persistence of fipronil and its primary metabolite fipronil sulfone after oral and dermal administration by simulating natural farming conditions. Fipronil and fipronil sulfone detection and quantification were performed in different poultry matrices using an LC-MS/MS method coupled with modified QuEChERS extraction. After oral administration, fipronil was detected in feathers at each sampling time, in eggs for 28 days, and in the internal organs at the end of the experiment. After dermal administration, high levels of fipronil were detected in feathers, accounting for 195.85 ± 8.54 mg/kg, which were reduced by a third after 60 days. No traces of fipronil were detected in the eggs or internal organs. In addition, fipronil sulfone showed remarkable residues in all samples in trial 2. The data obtained confirmed that inappropriate use of unauthorized pesticides can lead to severe contamination of entire poultry farms. The contemporary presence of fipronil sulfone in feathers and eggs associated with the lack of fipronil in eggs suggests recent dermal contamination or past oral contamination. Moreover, simultaneous analysis of hens' feathers and eggs could represent a new method to improve large-scale monitoring programs and animal welfare, limiting their slaughter.

Nanoliposomes@Transcutol for In Vitro Skin Delivery of 8-Methoxypsoralen.

8-methoxypsoralen is the most common drug in psoralen plus ultraviolet light irradiation therapy for the treatment of severe psoriasis. Despite of the efficacy, its classic oral administration leads to several serious adverse effects. However, the topical psoralen application produces a drug skin accumulation lower than that obtained by oral administration, due to the drug low skin permeability. In this paper, 8-methoxypsoralen loaded Penetration Enhancer-containing Vesicles were prepared using soy phosphatidylcholine and the penetration enhancer Transcutol® (5% or 10%) and characterized in terms of size, polydispersity index, zeta potential and encapsulation efficiency. No statistically significant differences in both size (~135 nm) and encapsulation efficiency (~65%) were found for different Transcutol® concentration. Transdermal delivery study assessed by Franz diffusion cells, showed that the 8-methoxypsoralen mainly accumulated into the stratum corneum. Moreover, after Penetration Enhancer-containing Vesicles application, the drug recovered in this layer is almost double of that delivered by conventional liposomes, while no significant difference was found from the different Transcutol® concentrations. Finally, biocompatibility checked by an MTT assay, demonstrated that the incubation of human keratinocytes for 24 h with 8-methoxypsoralen loaded Penetration Enhancer-containing Vesicles did not significantly reduce cell viability.

Zoxamide accumulation and retention evaluation after nanosuspension technology application in tomato plant.

BACKGROUND: Low water solubility of pesticide requires formulations with high levels of stabilizers and organic solvents. Moreover, only 0.1% of the applied pesticides formulation reaches the target, while 99.9% spreads in the surrounding environment. Therefore, there is the need for more efficient and environmentally sustainable alternatives. RESULTS: Zoxamide (ZO) nanosuspension was prepared through a media milling technique by using the stabilizer polysorbate 80. The thin and acicular crystals obtained, showed particle size of 227 nm, polydispersion index of 0.247 and zeta potential of -28 mV. Dimensional data and morphology of ZO nanocrystals alone, on tomato leaves and berries, were confirmed by scanning electron microscopy. The reduction in size for ZO crystals obtained after the milling process increased pesticide water solubility till 39.6 mg L-1 , about 1.6 the solubility obtained with a conventional commercial formulation. Field and dip contamination trials performed on tomato plants showed the nanosuspension's ability to increase ZO deposition and accumulation versus a coarse ZO suspension and commercial formulation, respectively. CONCLUSIONS: The nanoformulation proposed, resulted in low cost and was easy to make. Moreover, the organic solvent-free composition together with a low surfactant addition assured a minor environmental impact. Finally, the increased retention and deposition of the fungicide can reduce the amounts of ZO formulation applied to tomatoes. © 2021 Society of Chemical Industry.

Multi-platform metabolomic approach to discriminate ripening markers of black truffles (Tuber melanosporum).

Black truffle is characterized by a black ascocarp and white veins. This hypogeous fruit body is known for its aroma. Understanding metabolic variation during ripening can shed light on truffle biology. In this work, the comprehensive polar metabolome and the volatile organic compounds of T. melanosporum were studied at different ripening stages by means of a metabolomic approach using GC-MS. Multivariate statistical data analysis indicated that the metabolic profile changed during ripening and that the metabolites that mostly discriminated truffles in the early ripening stages belonged to the classes of carbohydrates, while free fatty acids and amino acids, among which precursors of VOCs, characterized the late stages of ripening. Principal component analysis of the volatilome indicated that dimethylsulfide and dimethyldisulfide characterized most of the samples collected in December-January, while 1-octen-3-ol samples collected in February-March.

Integrated environmental evaluation of heavy metals and metalloids bioaccumulation in invertebrates and seaweeds from different marine coastal areas of sardinia, mediterranean sea.

In this work, three gastropods Patella vulgata, Osilinus turbinata, and Tahis clavigera, one echinoderm Parancetrotus lividus, one coelenterate Anemonia sulcata, and two seaweed Padina pavonica, and Cystoseira mediterranea were collected from three different marine areas of Sardinia in the Mediterranean sea and studied for heavy metals and metalloid content and accumulation trends. Inductively coupled plasma optical emission spectrometry (ICP-OES) was used for the determination of Al, AS, B, Ba, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Te, Ti, V and Zn in the selected samples. The results showed that gastropods were capable of accumulating Al, Ba, Cu, Fe, Sr, and Zn; seaweeds can better concentrate Al, Fe, and Zn than all other species. At the same time, echinoderms and coelenterate had limited ability to store specific metals, showing a much more homogeneous distribution. PCA analysis allowed us to discriminate among the sites and the species. Cala Zafferano was the area with the higher values of accumulation of all metals in all species as expected, considering its proximity to industrial sites. The results of the analysis showed clearly that heavy metal and metalloid accumulation was different for each species studied. Therefore, for a correct environmental assessment of a given area, a comprehensive approach is strongly recommended by exploiting the different properties of both accumulation and concentration of the metals by different aquatic species.

Influence of the Technological Process on the Biochemical Composition of Fresh Roe and Bottarga from Liza ramada and Mugil cephalus.

Bottarga is a high-priced delicacy with high nutritional value, and, in Italy, bottarga from mullets has been recognized to be a traditional food product. The flathead grey mullet Mugil cephalus and the thinlip grey mullet Liza ramada are the main cultured grey mullets in the Mediterranean Sea. In this study, fresh roe and bottarga from these two species were investigated to evaluate the influence of the technological process and the species on their biochemical composition and health advantages. The 1 h/200 g salting-out step did not increase the levels of NaCl in the bottarga, although it highly decreased the levels of some heavy metals like Cu and Al. Processing of fresh roe in bottarga led to an essential modification of the lipid fraction, following a general series of monousatturated fatty acid (MUFA)> poliunsutturated fatti acid (PUFA) > saturated fatty acid (SAFA) and an increase in both ω3 and ω6 in Liza ramada. Moreover, bottarga showed higher levels of squalene and cholesterol and an increased Essential Amino Acid/Total Amino Acid ratio (EAA/TAA) in both species. In addition to the nutritional benefits for the consumer, the process proposed in this study may represent a reliable tool for local producers to obtain a final bottarga with both a reproducible biochemical composition and organoleptic characteristics.

Effects of Industrial Processing on Pesticide Multiresidues Transfer from Raw Tomatoes to Processed Products.

Pesticides are broadly used to improve food safety, although they can lead to adverse health effects on consumers. Various food processing approaches, at the industrial or domestic level, have been found to highly reduce the amount of pesticide residues in most food materials. In this work, samples of raw tomatoes were collected directly from the field and processed at the industrial level to produce purée, triple concentrated paste, fine pulp, and diced tomatoes. A multiresidue method based on a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged e Safe) sample preparation, followed by liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS) for the assessment of 116 pesticides residues, was used. The analytical method has been validated according to SANTE indications. The recovery yields ranged from 75.5% to 115.3%, repeatability (RSDr) ranged from 3.4% to 18.3%, while reproducibility (RSDwR) ranged from 5.4% to 19.8%. The limit of quantifications (LOQs) ranged from 2.35 µg kg-1 for benthiavalicarb to 6.49 µg kg-1 for allethrin. A total of 159 raw tomato samples were collected from the field. The analysis showed the presence of 46 pesticides with azoxystrobin and chlorantraniliprole the most represented. On the other hand, all industrially processed samples showed values ≤ LOD, confirming that post-harvest processes can lead to a decrease in pesticide residues from agricultural commodities.

Systemic Administration of Orexin a Loaded Liposomes Potentiates Nucleus Accumbens Shell Dopamine Release by Sucrose Feeding.

Orexin neurons originate in the lateral and dorsomedial hypothalamus and perifornical area and produce two different neuropeptides: orexin A (OxA) and orexin B (OxB), which activate OxR1 and OxR2 receptors. In the lateral hypothalamus (LH) orexin neurons are involved in behavior motivated by natural rewards such as palatable food (sugar, high-fat food) and it has been demonstrated similarly that the orexin signaling in the ventral tegmental area (VTA) is implicated in the intake of high-fat food. The VTA is an important area involved in reward processing. Given the involvement of nucleus accumbens (NAc) shell dopamine (DA) in motivation for food, we intended to investigate the effect of OxA on the basal and feeding-activated DA transmission in the NAc shell. OxA is a large peptide and does not cross the blood-brain barrier and for this reason was loaded on two kinds of liposomes: anti-transferrin-monoclonal antibodies (OX26-mAb) and lactoferrin-modified stealth liposomes. The effect of IV administration of both OxA liposomes on NAc shell DA was studied by microdialysis in freely moving rats. OxA, administered using both kinds of liposomes, produced a delayed and transitory increase in dialysate DA in the NAc shell, strongly and lastingly potentiated the increase in dialysate DA elicited by sucrose pellet consumption and increased the number of eaten pellets. These effects of OxA on DA transmission and feeding were prevented by the OxR1 antagonist SB 334867. Hence, OxA acting on VTA OxR1 can facilitate sucrose-stimulated NAc shell DA transmission directly by increasing the basal activity of VTA DA neurons that send their projections to the NAc shell.

Nile red nanosuspensions as investigative model to study the follicular targeting of drug nanocrystals.

The strategy of formulating poorly soluble actives as nanosuspension has been explored by more than a thousand research papers, with some medicinal products for oral and intravenous use having reached the market or advanced clinical trials. Interestingly, there is a limited number of reports of nanosuspensions formulated for dermal and transdermal drug delivery. In the present work, a nanocrystals suspension of the fluorescent, water-insoluble dye Nile Red, is prepared through a media milling technique and exploited to characterize the fate of the nanosuspended drug when applied on the skin. More in detail, the accumulation of Nile Red nanocrystals inside the hair follicles is evidenced by scanning electron microscopy, and the diffusion of drug molecules in the different skin layers is evaluated by confocal microscopy and skin permeation studies. Overall, the combination of the analytical techniques provide a description of the mechanisms underlying dermal accumulation, and transdermal penetration of a drug formulated as a nanosuspension.

Production of nanosuspensions as a tool to improve drug bioavailability: focus on topical delivery.

Over the past two decades nanosizing technology has become one of the most successful formulation approaches for improving the bioavailability of poorly soluble drugs, which show a low oral absorption due to low dissolution velocity. Nanocrystals are nanoparticles of pure drug, without any matrix material, with an average diameter below 1 µm (typically in the range of 200-500 nm), which can be prepared in both water and non-water media as colloidal nanosuspensions stabilized using surfactants or polymers. The reduction of the drug particle diameter below 1 µm increases the dissolution velocity by increasing the particle surface and decreasing the diffusion layer thickness. Nanosuspension production processes involve two different approaches such as bottom-up (drug nanocrystal precipitation) and top-down (drug particle disintegration) technologies or a combination of two. Within these main approaches, a variety of possible techniques to achieve particle size reduction have been proposed by different research groups from both industry and academia. Even though nanosuspensions formulations have been especially studied for oral and parenteral administration, nanocrystals have showed a great potential also for topical delivery through alternative routes such as dermal, pulmonary and ocular route. The purpose of this review is to describe the main technologies used to produce nanosuspensions as well as to explore the most significant results and progresses obtained by application of drug nanocrystal formulations through topical routes.

Maltodextrin fast dissolving films for quercetin nanocrystal delivery. A feasibility study.

The objective of this study was to evaluate the feasibility to prepare fast dissolving films as quercetin nanocrystal delivery systems, using maltodextrins as film forming material and glycerin as plasticizer, with the goal of enhancing quercetin oral bioavailability. Quercetin nanosuspensions were prepared using a high-pressure homogenizer, and then directly used to prepare the films by a casting method. Spectroscopic and calorimetric analysis evidenced that reduction of quercetin size at nanoscale and incorporation in maltodextrin films do not affect the solid state of the active ingredient. The loading of quercetin nanocrystals into the film determined a slight variation of film elasticity and ductility. Indeed, the elastic modulus of the loaded films resulted about a half of the placebo ones, while the elongation at break increased four folds. Free and film loaded quercetin nanocrystals showed a comparable dissolution rate, much higher than that of bulk quercetin.