LC-HR/MS Analysis of Lipophilic Extracts from Calendula arvensis (Vaill.) L. Organs: An Unexplored Source in Cosmeceuticals.

As part of a project aimed at promoting the use of Calendula arvensis (Vaill.) L. (field marigold, Asteraceae) phytocomplexes in cosmeceutical formulations, the chemical composition in apolar specialized metabolites is herein elucidated. Furthermore, the screening of the cytotoxicity of the apolar extracts was evaluated in order to underline their safety as functional ingredients for cosmetics. After dissection of Calendula organs (florets, fruits, leaves, bracts, stems, and roots), ultrasound-assisted maceration in n-hexane as an extracting solvent allowed us to obtain oil-like mixtures, whose chemical composition has been highlighted through a UHPLC-ESI-QqTOF-MS/MS approach. Twenty-nine metabolites were tentatively identified; different compounds, among which the well-known poly-unsaturated fatty acids, and oxylipins and phosphatides were detected for the first time in Calendula genus. The screening of the dose-response cytotoxicity of the apolar extracts of C. arvensis highlighted the concentration of 10 µg/mL as the most suitable for the formulation of cosmeceutical preparations. Sera enriched with leaf and fruit apolar extracts turned out to have the best activity, suggesting it can be used as a new source in skin care thanks to their higher content in fatty acids.

A Joint Approach of Morphological and UHPLC-HRMS Analyses to Throw Light on the Autochthonous 'Verdole' Chestnut for Nutraceutical Innovation of Its Waste.

Nowadays, chestnut by-products are gaining a lot of interest as a low-cost raw material, exploitable for developing added-value products. This is in line with suitable chestnut by-products' management, aimed at reducing the environmental impact, thus improving the chestnut industry's competitiveness and economic sustainability. In this context, with the aim of valorizing local cultivars of European chestnuts (Castanea sativa Mill.), our attention focused on the Verdole cultivar, which has been characterized by using the UPOV guidelines for its distinctness, homogeneity, and stability. After harvesting, Verdole chestnuts were properly dissected to collect the outer and inner shells, and episperm. Each chestnut part, previously crushed, shredded, and passed through diverse sieves, underwent ultrasound-assisted extraction. The extracts obtained were evaluated for their total phenolic, flavonoid, and tannin content. The antiradical capacity by DPPH and ABTS assays, and the Fe(III) reducing power, were also evaluated. Although all the samples showed dose-dependent antioxidant efficacy, plant matrix size strongly impacted on extraction efficiency. LC-HRMS-based metabolic profiling highlighted the occurrence of different polyphenol subclasses, whose quantitative ratio varied among the chestnut parts investigated. The outer shell was more chemically rich than inner shell and episperm, according to its pronounced antioxidant activity. The polyphenol diversity of Verdole by-products is a resource not intended for disposal, appliable in the nutraceutical sector, thus realizing a new scenario in processing chestnut waste.

Hemp Stem Epidermis and Cuticle: From Waste to Starter in Bio-Based Material Development.

Nowadays, hemp farmers are facing an urgent problem related to plant stem disposal after seed harvesting. In this work, the commonly discarded epidermis and cuticle of hemp stems were valorized, turning them towards a sustainable recycling and reuse, contributing to the circular economy concept. Cellulose deprived of amorphous regions was obtained by a green process consisting of an ethanolic ultrasound-assisted maceration followed by mild bleaching/hydrolysis. The obtained hemp cellulose was esterified with citric acid resulting in a 1.2-fold higher crystallinity index and 34 ∘C lower Tg value compared to the non-functionalized hemp cellulose. Green innovative biocomposite films were developed by embedding the modified cellulose into PLA by means of an extrusion process. The structural and morphological characterization of the obtained biocomposites highlighted the functionalization and further embedment of cellulose into the PLA matrix. Attenuated Total Reflectance-Fourier Transform Infrared spectroscopy (ATR-FTIR) results suggested physical and chemical interactions between PLA and the organic filler in the biofilms, observing a homogeneous composition by Field Emission-Scanning Electron Microscopy (FESEM). Moreover, some increase in thermal stability was found for biocomposites added with 5%wt of the hemp cellulose filler. The obtained results highlighted the feasible recovery of cellulose from hemp stem parts of disposal concern, adding value to this agro-waste, and its potential application for the development of novel biocomposite films to be used in different applications.

Cannabidiolic acid in Hemp Seed Oil Table Spoon and Beyond.

Cannabidiolic acid (CBDA) is the main precannabinoid in industrial hemp. It represents a common constituent of hemp seed oil, but mainly abundant in the aerial parts of the plant (including their processing waste). Thus, the optimization of fast and low-cost purification strategies is mandatory, as well as a deep investigation on its nutraceutical and cosmeceutical properties. To this purpose, CBDA content in hemp seed oil is evaluated, and its recovery from wasted leaves is favorably achieved. The cytotoxicity screening towards HaCaT cells, by means of MTT, SRB and LDH release assays, suggested it was not able to decrease cell viability or perturb cell integrity up to 10 µM concentration. Thus, the ability of CBDA to differentially modulate the release of proinflammatory cytokines and chemokines mediators has been evaluated, finding that CBDA decreased IFN-γ, CXCL8, CXCL10, CCL2, CCL4 and CCL5, mostly in a dose-dependent manner, with 10 µM tested concentration exerting the highest activity. These data, together with those from assessing antimicrobial activity against Gram(+) and Gram(-) bacteria and the antibiofilm formation, suggest that CBDA is able to counteract the inflammatory response, also preventing bacteria colonization.

Calendula arvensis (Vaill.) L.: A Systematic Plant Analysis of the Polar Extracts from Its Organs by UHPLC-HRMS.

Calendula arvensis (Vaill.) L. (field marigold, Asteraceae) is an alimurgic plant, whose flowers and leaves are a common part of local food dishes. The diversity in polar specialized metabolites is herein unraveled, with the aim to further promote and valorize the food use of the plant. To this purpose, following the plant dissection of its organs (florets, fruits, leaves, bracts, stems, and roots), ultrasound assisted maceration has been employed in order to recover phenols and polyphenols. Through an untargeted UHPLC-HR MS (Ultra-High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry) approach, and deeper investigation of the fragmentation patterns of each compound by tandem mass spectrometry, the florets' constitution in triterpene saponins and flavonol glycosides has been highlighted, whereas hydroxycinnamoyl compounds are mainly in bracts and fruits. The antiradical and reducing capabilities of the organs' extracts have been assessed, and data acquired have been analyzed by cluster analysis, which allowed bracts and fruits to be observed, despite their negligible food use, as the most active extracts. Chemical and antioxidant data on the diverse organs of field marigold suggest new investigative food and nutraceutical scenarios of this plant, also revalorizing and preserving its traditional uses.

Discrimination of CBD-, THC- and CBC-type acid cannabinoids through diagnostic ions by UHPLC-HR-MS/MS in negative ion mode.

The interest in industrial hemp-based products and by-products to be utilized in food and nutraceutical sector is strictly linked to the demand for improved analytical methods to rapidly discriminate acid phytocannabinoid isomers. Indeed, the differentiation of acid phytocannabinoids, also named pre-cannabinoids, is not properly exploited and valued until now, and it is challenging. Herein, using high-resolution MS/MS, the most common pre-cannabinoids with the resorcinol core linked to the alkyl five carbons chain were deeply investigated in terms of their reactivity to collision-induced dissociation, gaining key data on the integrated energy framework of their dissociation pathway. In fact, CBD-, THC- and CBC-type pre-cannabinoids could be discriminated based on the base peak identity, and the intensity of common fragment ions, when collision energy fragmented precursor ions by 70-75 %. In particular, energy-resolved CID mass spectra highlighted that fragmentation occurs, unrelatedly to alkyl chain length, at phenolic and monoterpenic moieties levels. Accordingly, this tool is effective for further differentiating pre-cannabinoid homologues, from methyl- up to heptyl-homologues, getting new insight in acid cannabinoids heritage of hemp and its products.

Polyphenols vs. Coronaviruses: How Far Has Research Moved Forward?

The epidemic, caused by SARS-CoV-2 at the beginning of 2020, led us to a serious change in our lifestyle that for about three months has confined us to our homes, far from our laboratory routine. In this period, the belief that the work of a researcher should never stop has been the driving force in writing the present paper. It aims at reviewing the recent scientific knowledge about in vitro experimental data that focused on the antiviral role of phenols and polyphenols against different species of coronaviruses (CoVs), pointing up the viral targets potentially involved. In the current literature scenario, the papain-like and the 3-chymotrypsin-like proteases seem to be the most deeply investigated and a number of isolated natural (poly)phenols has been screened for their efficacy.

(‒)-Cannabidiolic Acid, a Still Overlooked Bioactive Compound: An Introductory Review and Preliminary Research.

Cannabidiolic acid (CBDA) is the main phytocannabinoid in fiber and seed-oil hemp (Cannabis sativa L.) plants, but its potential health-related capabilities have been masked for years by a greater scientific interest towards its neutral derivative cannabidiol (CBD). This review aims to collect from the literature and critically discuss all the information about this molecule, starting from its biosynthesis, and focusing on its bioactivity, as an anti-inflammatory, anti-emetic, anti-convulsant, and anti-cancerogenic drug. Furthermore, in the awareness that, despite its multiple bioactive effects, currently poor efforts have been made to achieve its reliable purification, herein, we propose a relatively simple, fast, and inexpensive procedure for its recovery from pollen of industrial hemp cultivars. Spectroscopic and spectrometric techniques allowed us to unequivocally identify pure isolated CBDA and to distinguish it from the constitutional isomer tetrahydrocannabinolic acid (THCA-A).

Hempseed Lignanamides Rich-Fraction: Chemical Investigation and Cytotoxicity towards U-87 Glioblastoma Cells.

The weak but noteworthy presence of (poly)phenols in hemp seeds has been long overshadowed by the essential polyunsaturated fatty acids and digestible proteins, considered responsible for their high nutritional benefits. Instead, lignanamides and their biosynthetic precursors, phenylamides, seem to display interesting and diverse biological activities only partially clarified in the last decades. Herein, negative mode HR-MS/MS techniques were applied to the chemical investigation of a (poly)phenol-rich fraction, obtained from hemp seeds after extraction/fractionation steps. This extract contained phenylpropanoid amides and their random oxidative coupling derivatives, lignanamides, which were the most abundant compounds and showed a high chemical diversity, deeply unraveled through high resolution tandem mass spectrometry (HR-MS/MS) tools. The effect of different doses of the lignanamides-rich extract (LnHS) on U-87 glioblastoma cell line and non-tumorigenic human fibroblasts was evaluated. Thus, cell proliferation, genomic DNA damage, colony forming and wound repair capabilities were assessed, as well as LnHS outcome on the expression levels of pro-inflammatory cytokines. LnHS significantly inhibited U-87 cancer cell proliferation, but not that of fibroblasts, and was able to reduce U-87 cell migration, inducing further DNA damage. No modification in cytokines' expression level was found. Data acquired suggested that LnHS acted in U-87 cells by inducing the apoptosis machinery and suppressing the autophagic cell death.