INTRODUCTION Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. OBJECTIVE After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe-emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. METHODOLOGY Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO(2) extraction). Separation and quantification of anthraquinones were accomplished using a reversed-phase C(18) column with the mobile phase of H(2)O-methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. RESULTS All calibration curves were linear over the concentration range tested (10-200 mM) with the determination coefficients >or=0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. CONCLUSION The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays.

Several species belonging to the genus Rhamnus (Rhamnaceae), comprising ones among which are found the most typical plants of the Italian flora, are known to contain biologically active anthraquinone secondary metabolites. Although several Rhamnus species were so far investigated, no information is available concerning the content and relative abundances of anthraquinones in R. saxatilis. In this study we used a simple, reliable, and accurate analytical method to determine the anthraquinones in bark of R. saxatilis. This allowed us also to trace a comparative study on the efficacy of different extraction solvents in ultrasonication time dependent assays. Separation and quantification of anthraquinones were accomplished using a C18 column with the mobile phase of H2O:methanol (40:60, v/v, 1% formic acid) at a flow rate of 0.7mL/min and a detection wavelength of 254nm, while the qualitative analyses were also achieved at a wavelength of 435nm.Finally, the described HPLC method, was used to obtain a specific chemical fingerprint for this species in comparison with other species from the same family.

Potentilla detommasii Ten. (Rosaceae) is a perennial herb that grows on rocky gravels of South-Europe at altitude of 200–1200 meters [1]. No reports on phytochemical or ethnobotany properties of this species have been cited in the literature. The topic of our work was the isolation and characterization of secondary metabolites from the title plant collected at flowering stage. Extraction was performed by overnight maceration with increasing polarity solvents. The resulting extracts were repeatedly purified by SiO2 gel and HPLC chromatographies and each fraction obtained was analyzed by IR, UV and NMR spectroscopies, GC-MS and collecting data compared with those reported in the literature for the same compound whenever possible. The main secondary metabolites characterized were fatty acids,β-sitosterol, D-(+)-chiro-inositol and finally two pyrone-dicarboxylic acids, namely 2-pyrone-4,6-dicarboxylic acid 1 (R=H) and its homologous 5-methyl-2-pyrone-4,6-dicarboxylic acid 2 (R=CH3).

Three Italian strains of hyssop wildly grown in different natural habitats of the Abruzzi region (Central Italy) and classified as Hyssopus officinalis L. subsp. aristatus, were characterized on the basis of their essential oil composition. The oils were obtained by steam distillation of the fresh aerial parts of the plants and analyzed by GC and GC/MS. Thirty-three compounds were identified. Relevant differences in the quantitative composition were observed among the oils so that the existence of three different chemotypes could be realized. In particular, one of the strains was characterized by high contents of myrtenol (32.6%) and β-pinene (193%), another contained β-pinene (24.7%) and 1,8-cineole (231%) as main components, while the third was very rich in methyl eugenol (439%) and limonene (15.9%). Whereas the latter two strains were similar in composition to oils from Spain and Montenegro, respectively, the former possessed a very unusual oil composition. Key Word Index: Hyssopus officinalis subsp aristatus essential oil compositionchemotypesmyrtenolβ-pinene1,8-cineolemethyl eugenollimonene

Variability and genetic divergence of 11 field populations of seven species of the Campanula elatines and C. fragilis rock-plants group from the Mediterranean and pre-Alpine areas have been studied by starch-gel electrophoretic techniques. Campanula isophylla, C. elatines, C. elatinoides, C. fragilis subsp. fragilis and subs. cavolinii, C. garganica were collected in Italy, whilst C. fenestrellata subsp. istriaca and C. portenschlagiana came from W. Jugoslavia. Twelve enzymatic loci for each population were genetically analysed: PGI-1 and 2, PGM-2 IDH-1 and -2, SKDH, ME, ADH, GOT-2, MPI-1 and -2, SOD-1. The genetic distances among the above mentioned entities have been calculated by Nei's index and depicted in a dendrogram.

Riassunto Viene descritta una nuova specie del genere Goniolimon Boiss., nuovo per la Flora Italiana, rinvenuta in prati aridi calcarei, a circa 800 m s.l.m., nei pressi de L'Aquila, denominata Goniolimon italicum. Il suo corredo cromosomico è 2 n = 32.