Volatile Composition Variability of Arnica montana Wild Populations of Trentino-Alto Adige, Italy, Determined by Headspace-Solid Phase Microextraction.

Arnica montana is a plant distributed in most of Europe, including the Alpine arc and Apennines in Italy, and traditionally used worldwide for medicinal properties. Twelve natural populations of the species from Trentino-Alto Adige, Italy, were characterized using Headspace-Solid Phase Microextraction analysis for their volatile profile. Fifty-one compounds were detected in flower heads, the most abundant being (E)-Caryophyllene (23.4 %), 2,2,4,6,6-Pentamethylheptane (8.3 %), α- trans-Bergamotene (7.2 %), Germacrene D (5.7 %), and Hexanal (5.3 %). A multivariate analysis performed on the ten most abundant compounds grouped these investigated accessions into five main clusters. Three clusters, comprising together five accessions, were linked to the geographical origin of two collection sites. This work is a complete characterization of volatiles of the species by SPME analysis reported to date. Furthermore, results suggest that the species' volatile profile can be linked to the geographical origin of the natural populations and, therefore, represent a tool for evaluating biodiversity within the species.

Variability in the Essential Oil Composition of Achillea erba-rotta subsp. moschata (Wulfen) I. Richardson Growing in the Eastern Italian Alps.

Achillea erba-rotta subsp. moschata (Wulfen) I. Richardson is endemic of the Central Alps and distributed in several Italian Regions, where is used for its medicinal and aromatic properties. Nine wild populations of the species were characterized for their essential oil content and composition. In total 45 components were detected of which 40 were identified, accounting from 96.3 % to 98.8 % of the total oil. Monoterpenes represented the main class of compounds (from 84 % to 90.9 %), being cis-thujone (from 1.1 % to 48.4 %), camphor (from 1.8 % to 36.9 %), 1,8-cineole (from 4.9 % to 23.4 %), trans-thujone (from 2.2 % to 33.9 %) and p-cymene (from 1.8 % to 4.8 %) the most abundant ones. The relative amount of chamazulene, detected in some accessions, was here reported for the first time in this species. High variability in the essential oil composition resulted among the investigated accessions, which were grouped by multivariate analysis into four main clusters, in some cases according to their site of origin. Beyond their possible chemo-systematic significance, the obtained results represent useful indications for the selection of interesting chemotypes within the species, especially in the perspective of its future cultivation.

Non-Volatile Terpenoids and Lipophilic Flavonoids from Achillea erba-rotta Subsp. moschata (Wulfen) I. Richardson.

Musk yarrow (Achillea erba-rotta subsp. moschata (Wulfen) I. Richardson) is endemic to the Central Alps, and is used to flavour alcoholic beverages. Despite its popularity as aromatizing agent and its alleged beneficial effects on digestion, the phytochemical profile of the plant is still largely unknown and undiscovered. As a consequence, its authentication in aromatized products is impossible beyond sensory analysis allowing forgery. To address these issues, we phytochemically characterized a sample of musk yarrow from the Italian Eastern Alps, identifying, in addition to widespread phytochemicals (taraxasterol, apigenin), the guaianolides 3, 8, 9; the seco-caryophyllane 6; and the polymethoxylated lipophilic flavonoids 1, 4, and 5. The flavonoid xanthomicrol 1, a major constituent of the plant, was cytotoxic to HeLa cells, but only modestly affected primary 3T3 fibroblasts. On account of their stability, detectability by UV absorption, and concentration, the oxygenated flavonoids qualify as markers to validate the supply chain of the plant growers to consumers.

In vitro evaluation of the effects of methanolic plant extracts on the embryonation rate of Ascaridia galli eggs.

The present study aims to find efficient alternatives to synthetic anthelmintics among ethno-veterinary herbs. Ascaridia galli eggs isolated from the worm uterus were exposed in vitro to methanolic extracts (ME) of nine plant species such as Achillea millefolium (AM), Artemisia absinthium (AA), Artemisia vulgaris (AV), Cicerbita alpina (CA), Cichorium intybus (CI), Inula helenium (IH), Origanum vulgare (OV), Tanacetum vulgare (TV), Tanacetum parthenium (TP). Flubendazole (FL), 0.5% formalin with dimethylsulfoxide and Petri dishes without the addition of reagents were used as positive, negative and untreated control respectively. The effects of the different ME at concentrations 0.500, 0.325, 0.200 mg/ml were assessed on the embryonic development (ED) of the eggs in duplicate. Logit analysis was used to calculate EC50 values. A generalized linear mixed model, having plant species and concentration as fixed effect and day as repeated measure, was used to determine differences in ED. Estimated EC50 was the lowest for FL at 0.11 mg/ml. CA and TV followed with 0.27 mg/ml and 0.32 mg/ml. ED for FL was significantly lower (25%) than that of CA (47%). The analysis showed 0.5 mg/ml of the ME of CA and TV significantly affected the ED at 35% and 42% inhibitions respectively. The ED for all ME showed similar pattern i.e., relatively higher efficacy in the first experimental week compared to the rest of the experimental period. The effect from all multicomponent extracts is time and dose dependent. The plants have promising results in inhibiting ED, contributing to the identification of alternative anthelmintic treatments.

Assessment of the chemical and genetic variability among accessions of Cicerbita alpina (L.) Wallr., an alpine plant with anthelmintic properties.

Cicerbita alpina (L.) Wallr, is a perennial alpine plant and a member of the Asteraceae family, typically found at altitudes above 1000 meters in the Italian Alps. Although previously utilized primarily as a local delicacy, recent studies have revealed strong antiparasitic activity through in vitro experiments. In Europe, numerous chemical drugs employed to combat nematodes - helminths that infest the digestive tract of livestock - are banned due to their environmental harm or show only reduced efficiency because of the development of resistance. Consequently, there is a growing demand for new alternative anthelmintic treatments in agricultural practices. Specialized metabolites found in the extracts of C. alpina could offer a sustainable and biological alternative to chemical drugs, specifically for nematode control. For this purpose, a unique germplasm collection originating from eight distinct natural populations in the Italian Alps was analyzed for its chemical diversity using state-of-the-art targeted LC-MS/MS spectrometry, including quantification based on multiple reaction monitoring. The predominant metabolites identified from the species were the caffeic acid derivatives chicoric acid, chlorogenic acid, and 3. 5-dicaffeoylquinic acid, the sesquiterpene lactone derivative 8-O-acetyl-15-ß-D-glucopyranosyl lactucin and the flavone glycosides, apigenin-7-O-glucoside and luteolin-7-O-glucoside, alongside their precursors apigenin and luteolin, respectively. Additionally, the genetic diversity of eighty individual plants within the germplasm collection was evaluated using ten DNA molecular markers (Simple Sequence Repeats), successfully transferred from two closely related species (Cichorium intybus and Tanacetum parthenium). This investigation unveiled a significant range of genetic diversity within the examined populations, resulting in the establishment of three distinct genetic groups. The findings were further correlated with the original ecological environment and local climate conditions spanning a biennial period, indicating substantial variations among the different accessions and the intricate interplay between genetic background and environmental factors. These results could serve as a basis for future domestication of the species through plant breeding programs ensuring product quality, but also facilitating the cultivation of C. alpina in more diverse geographic regions.

Evaluation of the Farming Potential of Echinacea Angustifolia DC. Accessions Grown in Italy by Root-Marker Compound Content and Morphological Trait Analyses.

The Echinacea genus includes a number of species that are commercially employed for the preparation of herbal products. Echinacea angustifolia DC. is one of these and is widely used, mainly for its immunomodulating properties, as it contains a wide range of compounds that belong to different chemical classes. In particular, echinacoside, cynarin and lipophylic alkylamides are the main specialized metabolites of the roots and can be considered to be marker compounds. In this work, 65 E. angustifolia accessions have been compared in a field trial in Italy, with the aim of investigating the variability/stability of the weight and chemical composition of their roots in order to identify the accessions that are most promising for future genetic-improvement programs. The morphological characteristics of the aerial parts have also been investigated. Seventeen samples were discarded due to germination or plantlet-development issues. Seven of the remaining accessions were identified as being different Echinacea species after a combined phytochemical and morphological evaluation. The morphological traits of the epigeal part, the root weight and the chemical composition data of the 41 confirmed E. angustifolia accessions were submitted to multivariate statistical analysis and a moderately homogenous sample distribution, with low selected-marker variability, was observed. Good echinacoside content was detected in almost all roots (>0.5%). However, two groups of accessions stood out because of their interesting features: One group possessed small roots, but had a high concentration of marker compounds, while another had highly developed roots and a good amount of marker compounds. These accessions can therefore be exploited for future selection work.

Monoterpenoids from the traditional North Italian vegetable Aruncus dioicus (Walter) Fernald var. vulgaris (Maxim.) H.Hara (Rosaceae).

Investigations of young shoots of Aruncus dioicus (Walter) Fernald var. vulgaris (Maxim.) H.Hara (Rosaceae), collected from the wild and used as vegetables in alpine provinces of Italy, yielded eight monoterpenoids. Besides known compounds, aruncin A, aruncide A, and cimicifugolide, five previously undescribed substances, aruncins C, D, and E, and aruncides D and E, were identified. Based on results from the full synthesis of aruncin B, structures of aruncin A and aruncide A were revised. Structures were established by HR mass spectrometry and extensive 1D and 2D NMR spectroscopy and based on data from synthetic aruncin B. An HPLC-DAD-ESI-MS method was developed to investigate the distribution of the monoterpenoids in different organs of Aruncus dioicus var. vulgaris and in aerial parts of A. dioicus var. aethusifolius (H.Lév.) H.Hara [Syn.: Aruncus aethusifolius (H.Lév.) Nakai]. Preliminary bioactivity studies moreover indicated weak cytotoxicity for some of the compounds against human prostrate adenocarcinoma cells.

Seasonal variation in secondary metabolites of edible shoots of Buck's beard [Aruncus dioicus (Walter) Fernald (Rosaceae)].

Aruncus dioicus (Walter) Fernald is an herbaceous plant belonging to the Rosaceae family. In the northern regions of Italy young shoots of A. dioicus are collected and used as vegetable. No studies on the chemical composition of young shoots of A. dioicus have been reported so far. Thus, the aim of the present paper was to develop and validate a comprehensive chromatographic method allowing the identification and quantification of chemical compounds occurring in A. dioicus shoots harvested at different developmental stages. The results showed that shoots contain mainly polyphenols. Twenty-four compounds were detected. Caffeoylglucose derivatives were recognized as major compounds. It was also confirmed that shoots contain the cyanogenic compound prunasin at all investigated vegetation stages. It was shown that the prunasin content significantly increases during plant development, thus justifying the belief that after the development of the first green leaves, shoots should not be used for culinary purposes any longer.