Bioprospecting of Targeted Phenolic Compounds of Dictyota dichotoma, Gongolaria barbata, Ericaria amentacea, Sargassum hornschuchii and Ellisolandia elongata from the Adriatic Sea Extracted by Two Green Methods.

The content of bioactive compounds in four brown and one red algae from the Adriatic Sea (Dictyota dichotoma, Gongolaria barbata, Ericaria amentacea, Sargassum hornschuchii and Ellisolandia elongata) is explored. The efficiency of two different extraction methods viz. ultrasound-assisted extraction (UAE) and matrix solid-phase dispersion (MSPD) to obtain the extracts rich in phenolic compounds was compared. The effect of the extraction solvent to modulate the phenolic profile was assessed. In general, the mixture ethanol/water in an isovolumetric proportion showed the best results. The total phenolic content (TPC) and antioxidant activity (AA), as well as the individual polyphenolic profile, were evaluated for five target algae. TPC values ranged between 0.2 mg GAE/g (for E. elongata) and 38 mg GAE/g (for S. hornschuchii). Regarding the quantification of individual polyphenols by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, the presence of a high number of hydroxybenzoic acid derivatives (mainly of 3- and 4-hydroxybenzoic acids) in all species was noted. In G. barbata their concentrations reached up to 500 mg/kg. IC50 values (ABTS assay) ranged between 44 mg/L (for S. hornschuchii) and 11,040 mg/L (for E. elongata). This work contributes to the in-depth characterization of these little-explored algae, showing their potential as a natural source of phenolic compounds.

Phytochemical Analysis and Antiproliferative Activity of Ulex gallii Planch. (Fabaceae), a Medicinal Plant from Galicia (Spain).

The genus Ulex comprises thirteen accepted species of perennial shrubs in the family Fabaceae. In Galicia (Spain) many of these are considered spontaneous colonizing species, which are easy to establish and maintain. Among them, Ulex gallii Planch. is used in traditional medicine for the same anti-infective, hypotensive and diuretic purposes as Ulex europaeus L., which is the most studied species. Likewise, some studies have described the antitumoral properties of several species. However, there are few scientific studies that justify the use of Ulex gallii Planch. and nothing has been reported about its composition to date. In our study, the entire plant was extracted with methanol and the crude extract was subjected to liquid phase extraction with distinct solvents, yielding three fractions: hexane (H), dichloromethane (D) and methanol (M), which were subsequently fractionated. The dichloromethane (D5, D7 and D8) and methanol (M4) sub-fractions showed antiproliferative activity on A549 (lung cancer) and AGS (stomach cancer) cell lines, and caspase 3/7 activity assessment and DNA quantification were also performed. Targeted analysis via UHPLC-QToF, in combination with untargeted analysis via MS-Dial, MS-Finder and Global Natural Products Social Molecular Networking (GNPS), allowed us to tentatively identify different metabolites in these sub-fractions, mostly flavonoids, that might be involved in their antiproliferative activity.

Understanding the chemical and mineralogical composition of commercial henna and jagua tattoos and dyes-a multi-analytical approach.

Temporary tattoos and dyes constitute a great analytical challenge in relation to the regulatory control of their ingredients. Most of these commercial products are not labeled according to their content and their chemical nature is highly diverse. Therefore, it is necessary to analyze these complex samples to evaluate the potential presence of metallic impurities, to ensure the safety of cosmetic products contributing to health protection. This study proposes a multi-analytical methodology, which includes handheld X-ray fluorescence (h-XRF) and X-ray diffraction (XRD), complemented by variable pressure scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (VP-SEM-EDS) to fully characterize 34 commercial samples of jagua and henna tattoos and dyes. The approach allowed the identification of the main constituents providing complementary compositional data and differences between sample types were established. In addition, information on the degree of natural pigments homogeneity was also obtained. The results' discussion considering the current European cosmetics regulation may be useful to support the drafting of safety requirements and specific regulation.

Recent Advances in Sample Preparation for Cosmetics and Personal Care Products Analysis.

The use of cosmetics and personal care products is increasing worldwide. Their high matrix complexity, together with the wide range of products currently marketed under different forms imply a challenge for their analysis, most of them requiring a sample pre-treatment step before analysis. Classical sample preparation methodologies involve large amounts of organic solvents as well as multiple steps resulting in large time consumption. Therefore, in recent years, the trends have been moved towards the development of simple, sustainable, and environmentally friendly methodologies in two ways: (i) the miniaturization of conventional procedures allowing a reduction in the consumption of solvents and reagents; and (ii) the development and application of sorbent- and liquid-based microextraction technologies to obtain a high analyte enrichment, avoiding or significantly reducing the use of organic solvents. This review provides an overview of analytical methodology during the last ten years, placing special emphasis on sample preparation to analyse cosmetics and personal care products. The use of liquid-liquid and solid-liquid extraction (LLE, SLE), ultrasound-assisted extraction (UAE), solid-phase extraction (SPE), pressurized liquid extraction (PLE), matrix solid-phase extraction (MSPD), and liquid- and sorbent-based microextraction techniques will be reviewed. The most recent advances and future trends including the development of new materials and green solvents will be also addressed.

Multi-Target Strategy to Uncover Unexpected Compounds in Rinse-Off and Leave-On Cosmetics.

The wide range and complexity of cosmetic formulations currently available on the market poses a challenge from an analytical point of view. In addition, during cosmetics manufacture, impurities coming from raw materials or formed by reaction of different organic compounds present in the formulation may be present. Their identification is mandatory to assure product quality and consumer health. In this work, micro-matrix solid-phase dispersion (µMSPD) is proposed as a multi-target sample preparation strategy to analyze a wide number of unexpected families of compounds including polycyclic aromatic hydrocarbons (PAHs), pesticides, plasticizers, nitrosamines, alkylphenols (APs), and alkylphenol ethoxylates (APEOs). Analytical determination was performed by gas chromatography-mass spectrometry (GC-MS) for the determination of 51 target compounds in a single run, whereas liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed for the analysis of six APs and APEOs. Both methodologies were successfully validated in terms of linearity, accuracy, and precision in leave-on and rinse-off cosmetics. Limits of detection (LODs) were calculated in the low ng g-1, showing their suitability to determine trace levels of impurities and banned compounds with different chemical natures, providing useful tools to cosmetic control laboratories and companies.

Unraveling the environmental impacts of bioactive compounds and organic amendment from grape marc.

In a society that produces large amounts of solid waste, the search for new methods of valorisation has led to the development of techniques that make it possible to obtain new products from waste. In the case of bio-waste, biological treatment such as anaerobic digestion or composting appear to be suitable options for producing bio-energy or bio-fertilizers respectively. Vermicomposting is a method of converting solid organic waste into resources through bio-oxidation and stabilization of the organic waste by earthworms. The purpose of this study is to establish the environmental impacts of a complete route for the valorisation of grape pomace in order to identify environmental hotspots. In this valorisation route, different value-added products are produced with potential application in the cosmetic, food and pharmaceutical sectors. Priority was given to the use of primary data in the elaboration of the data inventories needed to perform the life cycle assessment (LCA). The main findings from this study reported that the energy requirement of the distillation process is an important hot spot of the process. Although the valorisation route has some poor results in terms of the two environmental indicators (carbon footprint and normalised impact index), when economic revenues were included in this analysis, its environmental performance was better than that of other alternatives for bio-waste recovery.

Strategies for recycling and valorization of grape marc.

Grapes are one of the most cultivated fruit crops worldwide. Either for wine or juice production, grape processing generates a large amount of residues that must be treated, disposed of or reused properly to reduce their pollution load before being applied to the soil. In this review, a special focus is given to the treatment and valorization of the winemaking by-product like grape marc via anaerobic digestion, composting and vermicomposting at laboratory, pilot, and industrial scales. The impact of the final products (digestates, composts, and vermicomposts) on soil properties is briefly addressed. Moreover, the role of grape marc and seeds as a valuable source of natural phytochemicals that include polyphenols and other bioactive compounds of interest for pharmaceutical, cosmetic, and food industries is also discussed. This is of paramount importance given the fact that sustainability requires the use of management and valorization strategies that allow the recovery of valuable compounds (e.g. antioxidants) with minimum disposal of waste streams.

Extreme cosmetics and borderline products: an analytical-based survey of European regulation compliance.

The cosmetic industry currently focuses on products with magnified or exaggerated effects or extremely long-lasting characteristics. There are also a number of related commercial products for which the regulatory framework is far from clear; they are called 'borderline', and the European authorities only recommend which regulations they need to comply with. In any case, all these products must be safe under reasonable conditions of use in accordance with the applicable laws in force in the European Union (EU) framework. In this context, adequate analytical methodology is needed to evaluate the degree of compliance. Ultrasound Assisted Extraction (UAE) procedures for the analysis of 70 cosmetic ingredients have therefore been developed in this work. Moreover, for cosmetics with plastic applicators, a Supported-UAE (Sup-UAE) method was also opportunely optimized to check if a partial transfer of plasticizers to the cosmetics-and thereby to the consumers-could happen. In a survey of 50 commercial products (30 'extreme' and 20 'borderline'), the methods afforded mean recoveries of about 100% and RSD values lower than 5% for UAE and 10% for Sup-UAE, and with detection limits far below the legal requirements, for all the target compounds, thereby demonstrating their analytical suitability. Results are discussed in detail for phthalates, fragrances (musks and allergens) and some frequent preservatives. Additionally, a labelling study was performed to check if the consumer is correctly and fully informed. Graphical abstract ᅟ.

Bioconversion of Scotch broom into a high-quality organic fertiliser: Vermicomposting as a sustainable option.

Wild Scotch broom ( Cytisus scoparius (L.) Link) shrubs are widely distributed throughout the world and, in some countries, are considered to be a threat to other plant species. The use of plant biomass from Scotch broom as a fertiliser seems to be the optimum solution for its disposal because it contains considerable amounts of macronutrients. However, its direct application to soils may cause phytotoxicity due to the release of polyphenols, which could negatively affect crop growth. This study evaluated the efficiency of vermicomposting in processing this leguminous plant on an industrial scale. Vermicomposting substantially reduced the biomass of Scotch broom (by 84%), mainly as a result of the loss of volatile solids. Simultaneously, the initial population of earthworms ( Eisenia andrei) increased remarkably throughout the process, offering the possibility of obtaining earthworm protein for animal feed. A nutrient-rich and stabilised peat-like material without polyphenol-associated phytotoxicity was obtained after 42 days of vermicomposting. Lower values of microbial biomass and activity, indicative of stabilised materials, were recorded at the end of the trial. These findings suggest that vermicomposting is an environmentally sound management system for Scotch broom and could easily be scaled up for industrial application.

Vermicomposting grape marc yields high quality organic biofertiliser and bioactive polyphenols.

Grape is the largest fruit crop in the world, and most (80%) of the harvested fruit is used to make wine. The main by-product of the wine industry is called grape marc, which consists of the stalks, skin, pulp and seeds that remain after pressing the grapes. The aim of this study was to evaluate whether grape marc could be processed by vermicomposting on an industrial scale to yield both a high-quality organic, polyphenol-free fertiliser and grape seeds (as a source of bioactive polyphenols). Vermicomposting reduced the biomass of grape marc substantially (by 58%), mainly as a result of the loss of volatile solids. After 2 weeks, the process yielded a nutrient-rich, microbiologically active and stabilised peat-like material that was easily separated from the seeds by sieving. Although the polyphenol content of the seeds was considerably reduced, this disadvantage was outweighed by the ease of separation of the seeds. Separation of the seeds also eliminated the polyphenol-associated phytotoxicity from the vermicompost. The seeds still contained useful amounts of polyphenols, which could be directly extracted for use in the pharmaceutical, cosmetic and food industries. The procedure described is effective, simple and economical, and could easily be scaled up for industrial application.

The generally recognized as safe (GRAS) microalgae Haematococcus pluvialis (wet) as a multifunctional additive for coloring and improving the organoleptic and functional properties of foods.

This work proposes the application of astaxanthin-rich H. pluvialis wet paste (HPW) as a partial substitute for wheat flour in the preparation of filloas, a dish that combines the basic ingredients of industrial bakery. The nutritional and color profile of HPW-enriched samples was evaluated by comparative analysis with a mixture of synthetic food dyes. The highest content of carotenoids (798 ± 12 µg g-1) and fatty acids (76 ± 2 mg g-1) was obtained for a filloa fortified with H. pluvialis in contrast to a non-significant dye response. Subsequently, the color stability of the fortified filloa was evaluated over time (3, 6 and 9 days), as well as its physicochemical properties and microbiological profile. As a result, HPW provided filloas with a longer shelf life, brightness (*L), and texture, in comparison with a mixture of synthetic dyes. In addition, an inhibitory effect of HPW towards mesophilic aerobic microorganisms in the food was obtained.

Antibacterial Activity against Clinical Strains of a Natural Polyphenolic Extract from Albariño White Grape Marc.

Infections caused by multidrug-resistant bacteria are becoming increasingly frequent and sometimes difficult to treat due to the limited number of antibiotics active against them. In addition, they can spread between countries and/or continents, which is a problem of great relevance worldwide. It is, therefore, urgent to find alternatives to treat infections caused by multidrug-resistant bacteria. This study aimed at exploring a possible therapeutic alternative in the fight against antibiotic resistance. Based on the known antibacterial capacity of polyphenols, we tested the antimicrobial activity of a polyphenolic extract of Albariño white grape marc on clinical strains since research on such bacteria has been very scarce until now. First, the extract was obtained using a medium-scale ambient temperature (MSAT) system, which is an efficient and sustainable extractive method. The determinations of the polyphenolic content of the extract and its antioxidant capacity showed good results. Using chromatographic and mass spectrometric tools, 13 remarkable polyphenols were detected in the extract. The antibacterial activity of our grape marc extract against nineteen clinical strain isolates, some of which are multidrug-resistant, was evaluated by means of the calculation of half of the maximum inhibitory concentration (IC50) and the value of the minimum bactericidal concentrations (MBCs). In conclusion, the extract showed effectiveness against all clinical strains tested, regardless of their level of antibiotic resistance, and shows promise in the fight against antibiotic resistance.

The chemistry behind the body art: unveiling the elemental profile and heavy metal content of natural tattoos and dyes by ICP-MS.

Temporary tattoos and dyes are increasing popularity as an alternative to permanent ones. However, knowledge on its elementary composition is scarce and, this scientific gap can potentially make them a source of heavy metal exposure on humans. The present research aimed to explore the possibility of using the inorganic chemical signature to characterize natural pigments-based products and to evaluate the heavy metal content in jagua temporary tattoos and henna temporary hair dyes and tattoos. Thirty-four different samples of commercial products were analyzed for the presence and quantity of eleven trace elements (Al, Cr, Mn, Co, Ni, Cu, Zn, As, Cd, Ba, and Pb) by inductively coupled plasma mass spectrometry (ICP-MS). The overall mean concentrations varied between 0.02 and 973 µg g-1 in solid samples or 0.01-1878 µg g-1 for paste ones, wherein potential fake products were uncovered. None of the 34 samples considered comply with the current European Cosmetics Regulation. Samples were differentiated according to their Cr, Zn, Ba, and Pb content. The overall results revealed highly variable levels in the investigated samples, which leads us to suggest detailed quality controls of these materials, due the fact that their continued use can origin potential threat to human health.

Optimization of bioactives extraction from grape marc via a medium scale ambient temperature system and stability study.

A scalable procedure with minimum energy requirements, MSAT (Medium Scale Ambient Temperature), in combination with solvents generally recognized as safe (GRAS), has been optimized to obtain polyphenolic extracts from white grape (Vitis vinifera) marc. The solvents considered were propylene glycol (Pg), ethanol (Et), and ethyl lactate (Lc), as well as their respective hydro-organic mixtures. In a first approach, the operating parameters were optimized through a response surface matrix: extraction solvent volume (range 10-150 mL), marc mass (range 20-200 g) and marc/dispersant mass ratio (range 0.5-2 g⋅g-1), using the total polyphenol content (TPC) and the antioxidant activity (AA) of the extracts as response parameters. The highest TPC (5,918 mgGAE⋅L-1) and AA (44 mmolTE⋅L-1) values were obtained using 200 g marc and 100 mL solvent. Regarding the type of solvent, a better response was reached with Lc > Et > Pg > H2O obtaining a polyphenol concentration of 252 mg⋅L-1 for the hydro-organic isovolumetric ratio of ethyl lactate. In addition, the stability of the extracts was studied for 62 days. The effect of factors such as temperature, light exposure, and oxidative reactivity was evaluated. The bioactivity indices showed no changes with the storage conditions of the extracts in the first month of analysis, after which 75% of the antioxidant activity as the concentration of the polyphenolic profile (204 mg⋅L-1) remains. The absence of reactive oxygen and the cooling of the extract (4°C) were the most determining factors (p < 0.05) in modulating the stability of the total polyphenolic profile.

Occurrence of Patulin and Polyphenol Profile of Croatian Traditional and Conventional Apple Cultivars during Storage.

Apples and apple-based products are among the most consumed fruits around the world. However, they are susceptible to infection with the fungi Penicilium expansum. In addition to the reduction of apple quality, secondary metabolism of this fungus produces a mycotoxin patulin that has a negative effect on human health. Currently, there is no available research in the literature on the resistance of Croatian traditional apple cultivars to contamination with P. expansum, and consequently, on the patulin content in apples and apple juice produced from those apples. Although the mechanism of apple resistance to fungal diseases has not yet been sufficiently investigated, some studies have shown that polyphenolic compounds have some impact on fungi growth. In order to contribute with new knowledge, this research deals with monitoring the growth of P. expansum on apples, patulin detection by LC/MS-MS, determination of polyphenol profile by validated HPLC method, and determining the effect of polyphenolic compounds on fungi growth and patulin production during apple storage. The results of this study have shown that Croatian traditional apple cultivars harvested from family farm Horvatic contain higher concentration of polyphenolic compounds and higher antioxidant activity. At the same time, they showed more resistance to infection by P. expansum than conventional ones. The higher content of dihydrochalcones and flavanols encouraged the biosynthesis of patulin in examined cultivars. However, the higher content of non-flavonoids such as 2-6 dimethoxybenzoic acid, 4-hydroxycinnamic acid and chlorogenic acid leads to decrease in content of patulin. In conclusion, it seems that content of polyphenols and patulin production are correlated.

Identification and Quantification of Polyphenols in Croatian Traditional Apple Varieties.

Apples and apple-based products are particularly interesting due to being a good source of polyphenols in an everyday diet. Recently there has been increased interest in the preservation of traditional apple varieties due to studies that suggest that traditional apple varieties have a higher content of polyphenols and antioxidant activity compared to commercial varieties. This study shows that traditional apple varieties contain higher concentration of polyphenolic compounds than conventional ones, such as chlorogenic acid (1.29-456 mg/kg dw), catechin (0.70-312 mg/kg dw), epicatechin (1.75-244 mg/kg dw), procyanidin B1 + B2 (4.08-358 mg/kg dw) and quercetin-3-glucoside (0.96-231 mg/kg dw). This research underlies the great potential of traditional apple varieties as a source of natural antioxidants and polyphenolics.

Analysis of multi-class preservatives in leave-on and rinse-off cosmetics by matrix solid-phase dispersion.

Matrix solid-phase extraction has been successfully applied for the determination of multi-class preservatives in a wide variety of cosmetic samples including rinse-off and leave-on products. After extraction, derivatization with acetic anhydride, and gas chromatography-mass spectrometry analysis were performed. Optimization studies were done on real non-spiked and spiked leave-on and rinse-off cosmetic samples. The selection of the most suitable extraction conditions was made using statistical tools such as ANOVA, as well as factorial experimental designs. The final optimized conditions were common for both groups of cosmetics and included the dispersion of the sample with Florisil (1:4), and the elution of the MSPD column with 5 mL of hexane/acetone (1:1). After derivatization, the extract was analyzed without any further clean-up or concentration step. Accuracy, precision, linearity and detection limits were evaluated to assess the performance of the proposed method. The recovery studies on leave-on and rinse-off cosmetics gave satisfactory values (>78% for all analytes in all the samples) with an average relative standard deviation value of 4.2%. The quantification limits were well below those set by the international cosmetic regulations, making this multi-component analytical method suitable for routine control. The analysis of a broad range of cosmetics including body milk, moisturizing creams, anti-stretch marks creams, hand creams, deodorant, shampoos, liquid soaps, makeup, sun milk, hand soaps, among others, demonstrated the high use of most of the target preservatives, especially butylated hydroxytoluene, methylparaben, propylparaben, and butylparaben.

Antimicrobial Activity of Polyphenols and Natural Polyphenolic Extracts on Clinical Isolates.

Antibiotic resistance is a growing global problem that affects people, animals, the environment, and the economy. Many clinically relevant bacteria have become resistant to antibiotics, and this fact is emerging as one of the major threats to public health. The lack of new antibiotics, which is due to their time-consuming and costly development, exacerbates the problem. Therefore, it is necessary to identify new antimicrobial agents to treat bacterial and fungal infections. Plant extracts, which are valuable sources of bioactive compounds, mainly polyphenols, play an important role as a new strategy to combat pathogenic microorganisms. There is an extensive body of supporting evidence for the potent antibacterial and antifungal activities of polyphenols. Furthermore, some polyphenols show a synergistic effect when combined with antibiotics and antifungals, suggesting a promising alternative for therapeutic strategies against antibiotic resistance. However, only a few articles are found when searching the antibacterial or antifungal activities of polyphenols employing clinical isolates. Hence, this review focuses on the antimicrobial activity of polyphenols and extracts rich in polyphenols on clinical isolates, organized according to the World Health Organization priority pathogens classification.

Multicomponent Polyphenolic Extracts from Vaccinium corymbosum at Lab and Pilot Scale. Characterization and Effectivity against Nosocomial Pathogens.

An extraction method was designed and scaled up to produce multicomponent polyphenolic extracts from blueberries (Vaccinium corymbosum) of three different varieties. The process was specifically drawn up to comply with green chemistry principles. Extracts were obtained for the direct assessment of their antimicrobial and antiadhesive activities, and their direct use in the control of infections caused by concerning multidrug-resistant nosocomial pathogens. Analytical characterization was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Similar qualitative profiles were obtained in the three studied varieties with some significant quantitative differences. Up to 22 different polyphenols were identified with a clear predominance of anthocyani(di)ns followed by flavanols, non-flavonoids, and far behind by flavan-3-ols and procyanidins. The individual content of the main polyphenols was also discussed. A pilot scale extract has been also produced as a proof-of-concept, showing that scaling-up triples the content of bioactive phytochemicals. The effect of the polyphenolic extracts was analyzed against seven multidrug-resistance bacterial species by performing biofilm formation and growth and killing curves assays. All the studied varieties showed antibacterial and antiadhesive activities, being the extract containing the highest concentration of bioactive polyphenols, the most active with a high bactericidal effect.

Polyphenolic content and bioactivities of Crataegus oxyacantha L. (Rosaceae).

Solid-Liquid Extraction (SLE) using solvent of different polarities (CHCl3, EtOAc, and n-BuOH) has been applied to leaves and fruits from (Crataegus oxyacantha L.), a deciduous shrub with an expected rich phytochemical profile. The total polyphenols content and the radical scavenging activity of each extract were evaluated. These extracts were analyzed by HPLC-DAD and rutin, quercetin-3-glucoside, caftaric and caffeic acid had been positively identified. The phytochemical study of the ethyl acetate extract of C. oxyacantha, led to the isolation and structural elucidation of quercetin (1); quercetin-3-O-ß-glucoside (2); epicatechin (3); naringenin (4), reported for the first time from this species except caffeic acid and epicatechin. These compounds were identified by 1D and 2D NMR combined analysis as well as by MS and UV.The antimicrobial activity of these extracts has also been tested, showing strong antibacterial activity-solvent dependent-against Gram positive bacteria. Additionally, bactericidal power was demonstrated in fruit extracts.