Formal analyses are fundamental for the definition of honey, a product representing specific territories and their changes: the case of North Tyrrhenian dunes (Italy).

Honey is a variegate matrix depending significantly on the floral origin, and it could become an important agri-food product to valorise specific territories. Being so diverse, different analytical techniques are necessary for its description. Herein we characterized the honey produced in one of the Italian sand dunes systems hosting beekeeping activities. In terms of floristic origin, phytochemical characterization, and sensory and colour analysis, honey collected in 2021 and 2022 was comparable. Honey was polyfloral, with several pollens from dune habitat plants classified as minor. The presence of the allochthonous Amorpha fruticosa L. and the ruderal Rubus fruticosus L. pollens in the category of the secondary pollens testifies the alteration of the park vegetation. The phytochemical profile was rich in polyphenols. Other interesting compounds were coumarine derivatives, likely attributable to resin-laden plants as rockroses, long chain hydroxyacids typical of royal jelly and nicotinic acid and its analogues (2-hydroxynicotinic acid and 2-hydroxyquinoline). The above-mentioned honey showed interesting features and was a good representation of the vegetation of this area. Our study pointed out the importance of relying on multiple analytical techniques for the characterization of honey and the advisability of a technical support toward beekeepers to correctly describe and valorise their product.

Multidisciplinary analysis of Italian Alpine wildflower honey reveals criticalities, diversity and value.

Wildflower honeys produced in mountain grasslands are an expression of the biodiversity of these fragile habitats. Despite its importance, the botanical origin of honey is often defined without performing formal analysis. The aim of the study was to characterize six wildflower mountain honeys produced in the Italian Alps with different analytic techniques (SPME-GC-MS, HPLC-Orbitrap, cicatrizing and antioxidant activity) alongside melissopalynological analysis and botanical definition of the production area. Even though the apiaries were in mountain grasslands rich in Alpine herbaceous species, the honey could be defined as rhododendron/raspberry unifloral or raspberry and rhododendron bifloral while the honey produced at the lowest altitude differed due to the presence of linden, heather and chestnut. The non-compliance of the honey could be due to habitat (meadows and pastures) fragmentation, but also to specific compounds involved in the plant-insect relationship, such as kynurenic acid, present in a high quantity in the sample rich in chestnut pollen. 255 volatile compounds were detected as well as some well-known markers of specific botanic essences, in particular chestnut, linden and heather, also responsible for most of the differences in aroma profiling. A high correlation between nicotinaldehyde content and percentage of raspberry pollen (r = 0.853, p < 0.05) was found. Phenolic acid and hydroxy-fatty acid were predominant in the chestnut pollen dominant honey, which presented the highest antioxidant activity and the lowest cicatrizing activity, while the flavonoid fraction was accentuated in one sample (rhododendron pollen prevalent), that was also the one with the highest effect on wound closure, although all samples had similar cicatrizing effects apart from the chestnut pollen dominant honey (lowest cicatrizing activity). Our study highlighted the difficulty of producing mountain wildflower honey and the importance of a thorough characterization of this product, also to encourage its production and valorisation.

Effectiveness of Different Analytical Methods for the Characterization of Propolis: A Case of Study in Northern Italy.

: Propolis is used as folk medicine due to its spectrum of alleged biological and pharmaceutical properties and it is a complex matrix not still totally characterized. Two batches of propolis coming from two different environments (plains of Po Valley and the hilly Ligurian-Piedmont Apennines) of Northern Italy were characterized using different analytical methods: Spectrophotometric analysis of phenols, flavones and flavonols, and DPPH radical scavenging activity, HPLC, NMR, HSPME and GC-MS and HPLC-MS Orbitrap. Balsam and moisture content were also considered. No statistical differences were found at the spectrophotometric analysis; balsam content did not vary significantly. The most interesting findings were in the VOCs composition, with the Po Valley samples containing compounds of the resins from leaf buds of Populus nigra L. The hills (Appennines) samples were indeed characterize by the presence of phenolic glycerides already found in mountain environments. HPLC-Q-Exactive-Orbitrap®-MS analysis is crucial in appropriate recognition of evaluate number of metabolites, but also NMR itself could give more detailed information especially when isomeric compounds should be identified. It is necessary a standardized evaluation to protect and valorize this production and more research on propolis characterization using different analytical techniques.

Quality Traits of "Cannabidiol Oils": Cannabinoids Content, Terpene Fingerprint and Oxidation Stability of European Commercially Available Preparations.

Cannabidiol (CBD)-based oil preparations are becoming extremely popular, as CBD has been shown to have beneficial effects on human health. CBD-based oil preparations are not unambiguously regulated under the European legislation, as CBD is not considered as a controlled substance. This means that companies can produce and distribute CBD products derived from non-psychoactive hemp varieties, providing an easy access to this extremely advantageous cannabinoid. This leaves consumers with no legal quality guarantees. The objective of this project was to assess the quality of 14 CBD oils commercially available in European countries. An in-depth chemical profiling of cannabinoids, terpenes and oxidation products was conducted by means of GC-MS and HPLC-Q-Exactive-Orbitrap-MS in order to improve knowledge regarding the characteristics of CBD oils. Nine out of the 14 samples studied had concentrations that differed notably from the declared amount, while the remaining five preserved CBD within optimal limits. Our results highlighted a wide variability in cannabinoids profile that justifies the need for strict and standardized regulations. In addition, the terpenes fingerprint may serve as an indicator of the quality of hemp varieties, while the lipid oxidation products profile could contribute in evaluation of the stability of the oil used as milieu for CBD rich extracts.

Comprehensive quality evaluation of medical Cannabis sativa L. inflorescence and macerated oils based on HS-SPME coupled to GC-MS and LC-HRMS (q-exactive orbitrap®) approach.

There are at least 554 identified compounds in C. sativa L., among them 113 phytocannabinoids and 120 terpenes. Phytocomplex composition differences between the pharmaceutical properties of different medical cannabis chemotype have been attributed to strict interactions, defined as 'entourage effect', between cannabinoids and terpenes as a result of synergic action. The chemical complexity of its bioactive constituents highlight the need for standardised and well-defined analytical approaches able to characterise the plant chemotype, the herbal drug quality as well as to monitor the quality of pharmaceutical cannabis extracts and preparations. Hence, in the first part of this study an analytical procedures involving the combination of headspace-solid-phase microextraction (HS-SPME) coupled to GC-MS and High Resolution Mass-Spectrometry LC-HRMS (Orbitrap®) were set up, validated and applied for the in-depth profiling and fingerprinting of cannabinoids and terpenes in two authorised medical grade varieties of Cannabis sativa L. inflorescences (Bedrocan® and Bediol®) and in obtained macerated oils. To better understand the trend of all volatile compounds and cannabinoids during oil storage a new procedure for cannabis macerated oil preparation without any thermal step was tested and compared with the existing conventional methods to assess the potentially detrimental effect of heating on overall product quality.

Extracts and compounds active on TRP ion channels from Waldheimia glabra, a ritual medicinal plant from Himalaya.

BACKGROUND: Waldheimia glabra (Decne.) Regel is a wild plant from the Himalayan Mountains, commonly known as Smooth Ground Daisy. This plant is traditionally used by local populations in religious rituals (incense) or in traditional herbal medicine to treat skin diseases, headache, joint pain and fever. In literature few data are available on the investigation of this aromatic plant. PURPOSE: The present work aims at deepening knowledge about the chemical composition of W. glabra extracts and incense, as well as its activity on TRP ion channels. METHODS: Extracts and incense of W. glabra were analyzed by using HS-SPME GC/MS, GC/MS and NMR analysis. Tests on the activity of W. glabra extracts and isolated compounds (+)-ludartin 1 and B-ring-homo-tonghaosu 2 on TRP channels were also performed. RESULTS: Some extracts and pure compounds from W. glabra showed an interesting activity in terms of efficacy and potency on rat TRPA1, an ion channel involved in several sensory mechanisms, including pungency, environmental irritation and pain perception. Activity is discussed and compared with that of other known TRPA1 natural agonists with different chemical structures. All compounds showed only a negligible inhibition activity on rat TRPM8 ion channel. CONCLUSIONS: Our findings demonstrate that W. glabra is involved in the receptor activation mechanism and therefore represents a new natural product potentially useful in pharmaceutical and agrifood research.

Screening of the chemical composition and bioactivity of Waldheimia glabra (Decne.) Regel essential oil.

BACKGROUND: This research aimed at improving knowledge as to the chemical composition and the antibacterial and anti-cancer activities of the essential oil of Waldheimia glabra, a wild plant from the Himalayan Mountains. RESULTS: The results obtained by GC-MS showed that spathulenol, 9-tetradecenol, thujopsene, α-thujone, santolina alcohol and terpinen-4-ol were the main constituents of Waldheimia glabra essential oil. These results were confirmed by HS-SPME GC-MS analysis that also reported high amounts of artemisia alcohol and camphor. Disc diffusion assay suggested a mild antibacterial activity against both Escherichia coli and Staphylococcus aureus. Finally, a dose-response correlation was observed between Waldhemia glabra essential oil concentration and viability of human breast adenocarcinoma cells MDA-MB-231 and MCF-7. CONCLUSIONS: Together with the GC-MS method, HS-SPME GC-MS proved to be a reliable technique to characterise the chemical composition of essential oil obtained from aromatic plants. Further studies will focus on W. glabra phytochemicals and their biological activity, in order to support traditional uses of the plant. © 2015 Society of Chemical Industry.

Volatile fingerprint of italian populations of orchids using solid phase microextraction and gas chromatography coupled with mass spectrometry.

The volatile fraction of Ophrys sphegodes Mill. subsp. sphegodes, Ophrys bertolonii subsp. benacensis (Reisigl) O. Danesch, E. Danasch & Ehrend. and Neotinea tridentata (Scop.) R.M. Bateman, Pridgeon & M.W. Case, three orchid species with different pollinator attraction strategies, sampled in vivo and in situ, were evaluated by headspace solid phase microextraction coupled with gas-chromatography and mass spectrometry. The results were compared with the volatile compounds emitted by flowering plant samples picked from the same populations of orchid species. Hydrocarbons, aldehydes, alcohols and terpenes were the major constituents of "in vivo" orchid scents and some distinctive differences in volatile metabolite composition were observed between Ophrys and Neotinea species. Moreover, the odour bouquets of the picked flowering plant samples were different from the in vivo ones and in particular different proportions of the various terpenes and an increase of α-pinene were observed. In conclusion HS/SPME GCMS proved to be a suitable technique for analyzing and distinguishing the volatile fingerprint of different orchid species, sampled in vivo and in situ in a non-disruptive way, with potentially great advantages for ecophysiological studies of rare and endangered species.

Secondary metabolites and antioxidant capacities of Waldheimia glabra (Decne.) Regel from Nepal.

BACKGROUND: The phenolic content, volatile compound fingerprint and antioxidant capacity of Waldheimia glabra (Decne.) Regel (Asteraceae), a wild plant from the Himalayan mountains used in Sherpa religious rituals and in traditional medicine, were determined for the first time to investigate its suitability as a source of natural antioxidants. RESULTS: The total phenolic content and antioxidant capacity of dried and powdered plants from two different altitudes (5000 and 5200 m above sea level) in Khumbu Valley, Nepal were estimated by the Folin-Ciocalteu and 2,2-diphenyl-1-picrylhydrazyl radical methods respectively. The volatile organic compound (VOC) fingerprints from leafing plants (in vivo and in situ VOC sampling) and from dried plant samples were evaluated by headspace solid phase microextraction. The results indicated that W. glabra was a good source of phenolics with antioxidant properties. A correlation between total phenolic content and antioxidant capacity was found (r² = 0.9111). Seychellene, a volatile compound that characterises patchouli oil, was detected for the first time in a species of the Asteraceae family. CONCLUSION: Waldheimia glabra was proved to be a good source of antioxidants, partially supporting its traditional uses. Further studies aimed at deepening knowledge on W. glabra bioactivity and its possible uses will be of interest, considering the lack of proper scientific awareness regarding these aspects.

The relationships between phenolic content, pollen diversity, physicochemical information and radical scavenging activity in honey.

Honey is rich in different secondary plant metabolites acting as natural antioxidants and contributing to human health. Radical scavenging activity (RSA) is related to antioxidant activity, while the correlation between the phenolic content and RSA is often weak. Consequently, exclusive information on phenolics is often insufficient to qualify the RSA and the health promoting effects of honey. The paper deals with a case study of honey samples originating from the alpine areas of Italy's Lombardia and Veneto regions and realized by standard physicochemical and statistical analytical methods. In pure honey, the total phenolic content and the RSA were measured in spectrophotometric tests with the 2,2-diphenyl-1-picrylhydrazyl (DPPH·) free radical and Folin-Ciocalteu assays, respectively. Melissopalynological data was used to qualify pollen diversity through rank-frequency curves separating the samples into two groups. On the basis of physicochemical data, the samples were analyzed through multivariate classification and ranking procedures resulting in the identification of an outlier. Elimination of the outlier produced a high correlation between the total phenolic content and RSA in the two pollen diversity groups. The case study suggests that, after disregarding outliers, the RSA activity can be satisfactorily qualified on the basis of phenolics with pollen diversity as a covariate.

Influence of environmental factors on composition of phenolic antioxidants of Achillea collina Becker ex Rchb.

Effects of environmental growth conditions on the antioxidant capacity, total phenolic content and composition of Achillea collina Becker ex Rchb. were investigated. Methanol extracts and infusions obtained from leaves and inflorescences of plants cultivated in the Italian Alps at two different altitudes (600 and 1050 m a.s.l.) were evaluated. Infusions exhibited the highest antioxidant capacity (1/IC(50) values from 4.35 ± 0.72 to 8.90 ± 0.74), total phenolic content (from 31.39 ± 4.92 to 49.36 ± 5.70 mg gallic acid equivalents (GAE) g(-1) DW), chlorogenic acid (from 9.21 ± 1.52 to 31.27 ± 6.88 mg g(-1) DW), 3,5-di-O-caffeoylquinic acid (from 12.28 ± 3.25 to 25.13 ± 1.99 mg g(-1) DW) and 4,5-di-O-caffeoylquinic acid (from 7.38 ± 1.01 to 12.78 ± 2.61 mg g(-1) DW) content. Climate (as influenced by altitude) was shown to be the main environmental factor influencing yarrow composition and properties. Leaf extracts from the higher experimental site showed a 2-4-fold increase of chlorogenic acid level. Achillea collina can be considered as a very good source of bioactive phenolic compounds, and growing it at high altitude may constitute an effective way to significantly enhance its quality for both medicinal and nutritional uses.

Antioxidant and cytoprotective properties of infusions from leaves and inflorescences of Achillea collina Becker ex Rchb.

Plants are the main source of molecules with antioxidant and radical scavenging properties that aid the natural defence systems of cells and may be involved in the preservation of human health, particularly preventing all the physiopathological conditions where oxidative damage is a hallmark. Achillea collina Becker ex Rchb. is a medicinal plant of the Achillea millefolium aggregate (yarrow) traditionally used, particularly in mountain areas, as an infusion or alcohol extract for its digestive, antiinflammatory, analgesic, antipyretic and wound healing properties. The aim of this study was to investigate the antioxidant capacity and cytoprotective activity against oxidative stress of infusions obtained from the leaves and inflorescences of Achillea collina Becker ex Rchb., assessed by chemical (free radical scavenging activity by DPPH and Folin Ciocalteu assay) and biological assays (in vitro model of cytotoxicity and lipid peroxidation in PC12 cells line). Infusions of leaves had the highest antioxidant properties and cytoprotective activity. The antioxidant capacity was significantly correlated with the total phenolic content but not with the cytoprotective profile. Achillea collina Becker ex Rchb. has good antioxidant and cytoprotective properties, suggesting further investigations on its chemical composition and potential health value, particularly for traditionally prepared infusions of leaves.