Pubblicazioni
Sono riportati i titoli e gli abstracts di pubblicazioni sperimentali riguardanti diverse piante medicinali, appartenenti ai generi: Aloe, Cataranthus, Citrus, Digitalis, Echinacea, Ephedra, Evodia, Glycyrrhiza, Hypericum, Pteleopsis, Vernonia e altri
1991 VAMPA G; BENVENUTI S – SEPARATION OF 18-ALPHA-GLYCYRRHETINIC AND 18-BETA-GLYCY RRHETINIC ACID BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHIC DENSITOMETRY – JOURNAL OF CHROMATOGRAPHY – n. volume 543 – pp. da 479 a 482 ISSN: 0021-9673 [Articolo in rivista (262)]
Abstract – A high-performance thin-layer chromatographic method with scanning densitometry was developed for the simultaneous separation and determination of 18-alpha- and 18-beta-glycyrrhetinic acid.
1991 G. VAMPA, S. BENVENUTI, M. MELEGARI . Determination of glycyrrhizin in pharmaceutical preparations an in licorice products by high-performance thin-layer chromatographic densitometry. – ACTA TECHNOLOGIAE ET LEGIS MEDICAMENTI, vol. 2, p. 87-93, ISSN: 1121-2098
1992 -ANTOLINI L; VAMPA G; BENVENUTI S; PECORARI P – CRYSTAL AND MOLECULAR-STRUCTURE OF 18-ALPHA-GLYCYRRHETINIC ACID – JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS II – n. volume 1 – pp. da 65 a 67 ISSN: 0300-9580 [Articolo in rivista (262) ]
Abstract – The crystal structure of 18-alpha-glycyrrhetinic acid (trans junction of the D/E rings) has been determined in order to compare its stereochemical features with those of the known 18-beta form (cis junction of the D/E rings). The two stereoisomers exhibit different physiological actions. A comparison of their common moieties shows significant differences between the conformations of the unsaturated C ring, with some torsion angles differing by more than ten degrees. Further differences involve the convexity of the ABCD region, which in the alpha-form is smaller than that observed in the beta-isomer, and smaller than that reported for the mean steroid configuration.
1992 – Vampa, G; Benvenuti, S; Rossi, T. – Determination of glycyrrhizin and 18 alpha-, 18 beta-glycyrrhetinic acid in rat plasma by high-performance thin-layer chromatography. – IL FARMACO – n. volume 47(5 Suppl) – pp. da 825 a 832 ISSN: 0014-827X [Articolo in rivista (262)]
Abstract – An HPTLC method is described for the simultaneous determination of glycyrrhizinic acid tripotassium salt (3K-G), 18 alpha- and 18 beta-glycyrrhetinic acid (18 alpha-GA, 18 beta-GA) in rat plasma, after oral administration of 3K-G (30 mg/kg once a day for 30 days). Systolic pressure and the volume of urine excreted in 24 hours were recorded during this period to observe any drug-induced effects.
1993 – Zani, F; Cuzzoni, Mt; Daglia, M; Benvenuti, S; Vampa, G; Mazza, P. ( 1993 ) – Inhibition of mutagenicity in Salmonella typhimurium by Glycyrrhiza glabra extract, glycyrrhizinic acid, 18 alpha- and 18 beta-glycyrrhetinic acids – PLANTA MEDICA – n. volume 59(6) – pp. da 502 a 507 ISSN: 0032-0943 [Articolo in rivista (262)]
Abstract – The effects of Glycyrrhiza glabra L. extract, glycyrrhizinic acid, 18 alpha- and 18 beta-glycyrrhetinic acids on the mutagenicity of the ethyl methanesulfonate, N-methyl-N’-nitro-N-nitrosoguanidine, and ribose-lysine Maillard model systems were investigated by using the Salmonella/microsome reversion assay. The protocol used allowed us to detect desmutagenic and antimutagenic activity and to avoid false positive results due to toxicity. For all the compounds tested, no desmutagenic activity was observed against ethyl methanesulfonate and N-methyl-N’-nitro-N-nitrosoguanidine; only Glycyrrhiza glabra extract showed antimutagenic activity against ethyl methanesulfonate. On using the ribose-lysine mutagenic browning mixture, the desmutagenic activities of the Glycyrrhiza glabra extract, glycyrrhizinic acid, 18 alpha- and 18 beta-glycyrrhetinic acids were observed. 18 beta-Glycyrrhetinic acid was the most active compound. Glycyrrhiza glabra extract also exhibited antimutagenic activity against ribose-lysine
1994 – Rossi T; Vampa G; Benvenuti S; Pini L A; Galatulas I; Bossa R; Castelli M; Ruberto AI; Baggio G. – Glycyrrhizin and 18 beta-glycyrrhetinic acid: a comparative study of the pharmacological effects induced in the rat after prolonged oral treatment. – IN VIVO – n. volume 8 – pp. da 317 a 320 ISSN: 0258-851X [Articolo in rivista (262)]
Abstract – Recent clinical and toxicological studies have investigated the mineralcorticoid-like and hypertensive effects of liquorice, and we therefore set out to identify the active component responsible for these effects. We conducted a 30-day comparative analysis of glycyrrhizin and 18 beta-glycyrrhetinic acid and found that the latter causes significant variations both in systolic blood pressure and in the excretion in the urine of Ca++. The effects were fully reversible on suspension of treatment.
1995 – Benvenuti; F. Severi; L. Costantino; M. Melegari; A. Mucci – Identificazione, caratterizzazione e proprietà biologiche di derivati calconici da GLYCYRRHIZA GLABRA L. ( Convegno di fitochimica GENZIANA e SPECIE AMARO-AROMATICHE – Ricerche e Applicazioni – Camerino – 8-10 giugno 1995) ( – Convegno di fitochimica GENZIANA e SPECIE AMARO-AROMATICHE – Ricerche e Applicazioni – ATTI ) (Università degli Studi di Camerino Camerino ITA ) – n. volume unico [Abstract in Atti di convegno (27]
Abstract – Sono state isolate, purificate e caratterizzate diverse sostanze di natura calconica e ne è stata saggiata l’attività sull’Aldoso Reduttasi.
1996 -S.BENVENUTI, F.SEVERI,, L.COSTANTINO, M.MELEGARI, A.MUCCI – Identificazione, caratterizzazione e proprietà biologiche di derivati calconici da Glycyrrhiza glabra L. RIVISTA ITALIANA EPPOS, vol. 20, p. 13-16, ISSN: 0392-0445
1997 Benvenuti; C. Rustichelli; G. Vampa; V. Ferioli; G. Grassi; M. Melegari; P. Codeluppi; G. Gamberini – Analytical investigations on active principles in aloe leaf exudate ( 7th Meeting on Recent Development in Pharmaceutical Analysis RDPA ’97 – Isola d’Elba – 16-20 settembre 1997) ( – 7th Meeting on Recent Development in Pharmaceutical Analysis RDPA ’97 ) (Università di Firenze Firenze ITA ) – n. volume 1 – pp. da 91 a 91 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – The present work deals with the development of various procedures to estimate aloin and related compounds in aloe products by means of UV, HPTLC and HPLC procedures.
1998 – SEVERI F., BENVENUTI S., COSTANTINO L., VAMPA G., MELEGARI M, ANTOLINI L. . Synthesis and activity of a new series of chalcones as aldoso reducatse inhibitors. EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, vol. 33, p. 859-865, ISSN: 0223-5234
2000 – Fornasiero RB; Maffi L; Benvenuti S; Bianchi A – Morphological and phytochemical features of secretory structures in Hypericum richeri (Clusiaceae) – NORDIC JOURNAL OF BOTANY – n. volume 20 – pp. da 427 a 434 ISSN: 0107-055X [Articolo in rivista (262)]
Abstract – Structural and complementary chemical studies were carried out on Hypericum richeri, a lesser known species amongst those reported for folk medical use. We found only one type of secretory glands consisting of black dots which are present even in early emerging leaves. In the fully expanded leaves the nodular structure appears to be composed by a cluster of cells. These become unfunctional and disassembled towards the end of their development, and are used only as reservoirs of secretion products. HPTLC analyses showed that flower buds and flowers are the plant parts richest in active compounds. However, the spectrum of active compounds accumulated by H. richeri was both quantitatively and qualitatively similar to those reported for the pharmaceutically utilized. H. perforatum, and thus could potentially represent a possible alternative to this species.
2001 L.MAFFI, S.BENVENUTI, R.B.FORNASIERO, A.BIANCHI, M.MELEGARI – Inter-population variability of secondary metabolites in Hypericum spp. (Hypericaceae) of the Northern Apennines, Italy. NORDIC JOURNAL OF BOTANY, vol. 21, p. 585-593, ISSN: 0107-055X
Abstract – Hypericum spp. flowers were collected from different sites of Northern Apennines in Italy and were analysed by HPTLC technique to determine their contents in flavonoids (rutin, hyperosid, quercitrin, isoquercitrin and quercetin) and hypericins (pseudohypericin and hypericin). The H. pet bratum and H. richeri composition was evaluated in order to investigate the influence of ecological conditions on secondary metabolites production. The species Studied showed some differences in the amounts of investigated metabolites and high variability was observed in populations gathered in different places. No significant effects of environmental factors on active compound production were observed, except for the rutin content which is in positive correlation with the altitude of the growing site. Interesting informations are also given about the morphology of secretory structures found in Hypericum richeri.
2001 – Pellati; S. Benvenuti; M. Melegari; F. Firenzuoli – Separation and determination of adrenergic agonists from extracts and herbal products of Citrus aurantium L. var. amara by HPLC techniques ( 9th International Meeting on Recent Developements in Pharmaceutical Analysis (RDPA 01) – Lipari, Italy – 5-8 Giugno 2001) ( – 9th International Meeting on Recent Development in Pharmaceutical Analysis (RDPA 01) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Messina ITA ) – n. volume 1 [Abstract in Atti di convegno (274]
Abstract – Separation and determination of adrenergic agonists from extracts and herbal products of Citrus aurantium L. var. amara by HPLC techniques.
2002 F.PELLATI, S.BENVENUTI, M.MELEGARI, F.FIRENZUOLI – . Determination of adrenergic agonists from extracts and herbal products of Citrus aurantium L. var. amara by LC. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, vol. 29, p. 1113-1119, ISSN: 0731-7085
Abstract -The purpose of this study was to set up a HPLC method to separate adrenergic amines (dl-octopamine, dl-synephrine and tyramine) and to determine their content in fruits, extracts and herbal products of Citrus aurantium L. var. amara. A rapid method for the quantitative analysis of these amines is described. based on their separation by RP-HPLC technique with UV detection. The analysis were conducted on a Lichrospher RP-18 column at room temperature, using a mobile phase consisting of 0.02 M citric acid-0.02 M NaH2PO4 (7:3 v/v) and adjusted to a final pH of 3. The detection was at 220 nm. Since some of these amines are chiral compounds and their enantiomers showed different pharmacological activity, the direct separation of synephrine enantiomers was carried out with HPLC on a beta-cyclodextrin stationary phase, The mobile phase consisted of methanol-NaH2PO4 25 mM pH 3.5 (20:80 v/v) and tetrabutylammonium hydrogen sulfate 10 mM in ratio of 30:70 v/v in isocratic condition and the detection was at 220 nm. The two proposed methods were applied to the analysis of fruits, extracts and herbal products of C. aurantium L. var. amara. Taking into account that some authors have reported that l-synephrine may be converted into its d-form by high temperature, this optical isomerization was monitored by the same HPLC method used for the separation of enantiomers.
2002 – Pellati; S. Benvenuti; M. Melegari; L. Maffi; A. Bianchi – Analisi dicomponenti attivi e morfologia di Echinacea spp. ( 11° Congresso Nazionale della Società Italiana di Fitochimica – Riolo Terme, Italy – 6-9 Ottobre 2002) ( – 11° Congresso Nazionale della Società Italiana di Fitochimica ) (Società Italiana di Fitochimica Riolo Terme ITA ) – n. volume 1 [Abstract in Atti di convegno (274)]
Abstract – In questo studio, sono stati messi a punto metodi analitici avanzati e teniche di indagine farmacognostica per la caratterizzazione fitochimica di specie del genere Echinacea.
2003 – MAGRO L., DE TOGNI H.,, PELLATI F., BENVENUTI, S., MELEGARI M. – . Caratteristiche fitochimiche, proprietà farmacologiche e metodologie estrattive-analitiche di Vernonia kotchyana: recenti acquisizioni. PIANTE MEDICINALI, vol. 2, p. 249-254, ISSN:
2003 – Magro L.; De Togni H.; Pellati F.; Benvenuti; S. Melegari M. – Caratteristiche fitochimiche, proprietà farmacologiche e metodologie estrattive-analitiche di Vernonia kotchyana: recenti acquisizioni – PIANTE MEDICINALI – n. volume 2 – pp. da 249 a 254 ISSN: 1825-5086 [Articolo in rivista (262)]
Abstract – In questo articolo vengono descritte le recenti acquisizioni relative alle caratteristiche fitochimiche, proprietà farmacologiche e alle metodologie estrattive-analitiche applicate alla specie Vernonia kotchyana.
2003 – F.Pellati; S. Benvenuti; H. De Togni; L. Magro; M. Melegari; F. Soragni – Analysis of phenolic compounds and radical scavenging activity of Echinacea spp. ( 10th International Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2003) – Cogne, Italy – 28 Giugno-1 Luglio 2003) ( – 10th International Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2003) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Cogne ITA ) – n. volume 1 – pp. da 101 a 102 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this study, the phytochical analysis of phenolic compounds and radical scavenging activity of Echinacea spp. was described.
2004 – Benvenuti; F. Pellati; M. Melegari – Poly(ethyleneglycol) stationary phase for the analysis of natural bioactive compounds ( 15th International Symposium on Pharmaceutical and Biomedical Analysis (PBA 2004) – Firenze – 2-6 Maggio 2004) ( – 15th International Symposium on Pharmaceutical and Biomedical Analysis (PBA 2004) Abstract book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Firenze ITA ) – n. volume 1 – pp. da 423 a 423 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this study, the development and validation of a HPLC method for the analysis of bioactive compounds in H. perforatum extracts and natural products using a poly(ethyleneglycol) stationary phase was described.
2004 – F.Pellati; S. Benvenuti; D. Bertelli; F. Yoshizaki; M.C. Rossi – HS-SPME-GC-MS analysis of the volatile compounds of Evodia species fruits ( 25th International Symposium on Chromatography (ISC04) – Parigi – 4-8 Ottobre 2004) ( – 25th International Symposium on Chromatography (ISC04) Abstract Book ) (ISC04 Parigi FRA ) – n. volume 1 – pp. da 145 a 145 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this study, a HS-SPME-GC-MS method for the analysis of the volatile compounds of Evodia species fruits was develope
2004 – F.Pellati; S. Benvenuti; M. Melegari- Enantioselective LC analysis of adrenergic amines of natural origin on a protein-based chiral stationary phase ( 15th International Symposium on Pharmaceutical and Biomedical Analysis (PBA 2004) – Firenze – 2-6 Maggio 2004) ( – 15th International Symposium on Pharmaceutical and Biomedical Analysis (PBA 2004) Abstract book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Firenze ITA ) – n. volume 1 – pp. da 156 a 156 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this research, the enantioselective LC analysis of adrenergic amines of natural origin on a protein-based chiral stationary phase was carried out.
2004 – F. Pellati; S. Benvenuti; M. Melegari – Chromatographic performance of a new polar poly(ethyleneglycol) bonded phase for the phytochemical analysis of H. perforatum L. ( 25th International Symposium on Chromatography (ISC04) – Parigi – 4-8 Ottobre 2004) ( – 25th International Symposium on Chromatography (ISC04) Abstract Book ) (ISC04 Parigi FRA ) – n. volume 1 – pp. da 144 a 144 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract –In this study, the chromatographic performance of a new polar poly(ethyleneglycol) bonded phase was evaluated for the phytochemical analysis of H. perforatum L. plant material and commercial products.
2004 – F. PELLATI, S. BENVENUTI, M.MELEGARI . High-performance liquid chromatography methods for the analysis of adrenergic amines and flavanones in Citrus aurantium L. var. amara. PHYTOCHEMICAL ANALYSIS, vol. 15, p. 220-225, ISSN: 0958-0344
Abstract – Reverse-phase HPLC coupled with photodiode array detection was used for the simultaneous separation and determination of naturally occurring adrenergic amines (octopamine, synephrine and tyramine) in fruits and dry extracts of Citrus aurantium L. var. amara and in herbal medicines derived therefrom. Synephrine was the main component in fruits (0.10-0.35%) and in dry extracts (3.00-3.08%) and was present in the range 0.25-0.99% in herbal medicines. Flavanones were analysed in the same samples using a reverse-phase HPLC technique which allowed the identification and quantification of neoeriocitrin, narirutin, naringin, hesperidin, neohesperidin, naringenin and hesperetin. C. aurantium fruits and derivatives contained mainly glycosylated flavanones: in particular, naringin and neohesperidin were found to be the major flavonoids and their concentrations ranged from 1.80 to 26.30 and from 3.90 to 14.71 mg/g, respectively. The levels of aglycones were very low in all samples tested.
2004 – F. PELLATI, S.BENVENUTI, L.MAGRO, M.MELEGARI, F.SORAGNI . Analysis of phenolic compounds and radical scavenging activity of Echinacea spp.. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, vol. 35, p. 289-301, ISSN: 0731-7085
Abstract – The aim of this study was to set up and validate an RP-LC method with DAD-detection to quantify caffeic acid derivatives in various Echinacea spp. Samples were extracted with 80% methanol. The analyses were carried out on a Lichrospher RP-18 column (125 mm x 4 mm i.d., 5 mum), with a mobile phase gradient, which increases the acetonitrile level in a phosphoric acid solution (0.1%). The flow rate was 1.5 ml/min. Detection was set at 330 nm. This method allowed the identification and quantification of caftaric acid, chlorogenic acid, caffeic acid, cynarin, echinacoside and cichoric acid in Echinacea roots and derivatives. The total phenolic content was 10.49 mg/g for E. angustifolia, 17.83 mg/g for E. pallida and 23.23 mg/g for E. purpurea. Among Echinacea commercial herbal medicines, a certain variability in the concentrations of phenolic compounds was observed. The radical scavenging activity of Echinacea methanolic extracts was evaluated in vitro with a spectrophotometric method based on the reduction of an alcoholic 2,2-diphenyl-1-picrylhydrazyl (DPPH.) radical solution at 517 nm in the presence of a hydrogen donating antioxidant. As for pure compounds, echinacoside had the highest capacity to quench DPPH. radicals (EC50 = 6.6 muM), while caftaric acid had the lowest (EC50 = 20.5 muM). The average EC50 values for E. purpurea, E. pallida and E. angustifolia were 134, 167 and 231 mug/ml, respectively. The radical scavenging activity of Echinacea root extracts reflected their phenolic composition. The results indicate that Echinacea roots and derivatives are a good source of natural antioxidants and could be used to prevent free-radical-induced deleterious effects.
2004 – FAVALI M.; MUSETTI R.; S. BENVENUTI; BIANCHI A.; PRESSACCO L. – Catharanthus roseus L. plants and explants infected with phytoplasmas: alkaloid production and structural observations. PROTOPLASMA – n. volume 223 – pp. da 45 a 51 ISSN: 0033-183X [Articolo in rivista (262)]
Abstract – The results of several experiments concerning the presence and composition of alkaloids in different tissues (stems, leaves, roots) of Catharanthus roseus L. plants and explants, healthy and infected by clover phyllody phytoplasmas, are reported. The alkaloids extracted and determined by the reverse phase high-pressure liquid chromatography were vindoline, ajmalicine, serpentine, vinblastine, and vincristine. The total alkaloid concentration was higher in infected plants than in the controls, in particular the increase of vinblastine in infected roots was very significant. The ultrastructural observations of infected roots showed alterations of the cell walls and of the nuclei. These results demonstrate that phytoplasmas, detected in all infected tissues by light fluorescence and transmission electron microscopy, play an important role on secondary metabolism of the diseased plants, modifying both the total content of alkaloids and their ratio.
2005 – F.PELLATI, S.BENVENUTI, M.MELEGARI, T. LASSEIGNE . Variability in the composition of anti-oxidant compounds in Echinacea species by HPLC. PHYTOCHEMICAL ANALYSIS, vol. 16, p. 77-85, ISSN: 0958-0344
Abstract – A fast and reliable HPLC method for the determination of caffeic acid derivatives (caftaric acid, chlorogenic acid, caffeic acid, cynarin, echinacoside and cichoric acid) in various species of the genus Echinacea has been developed. Extraction of root samples by magnetic stirring with 80% methanol aqueous solution at room temperature allowed the complete recovery of all compounds of interest. Root extracts were analysed on a reversed-phase column with gradient elution and photodiode array detection. Caffeic acid derivatives showed differential qualitative and quantitative distributions in Echinacea species. The total amount of phenolic compounds ranged from 33.95 to 0.32 mg/g. The highest contents of caffeic acid derivatives were found in E. paradoxa var. paradoxa, E. paradoxa var. neglecta and E. purpurea, followed by E. angustifolia var. angustifolia, E. simulata, E. pallida and E. laevigata, whilst E. tennesseensis, E. sanguinea and E. atrorubens had low amounts of phenolic compounds. The radical scavenging activities of methanolic extracts of roots of Echinacea species was evaluated in vitro using the DPPH* radical scavenging method. The EC50 values of the samples ranged from 122 to 1223 mu g/mL. The radical scavenging activities of the root extracts were correlated with the content of phenolic compounds, with a correlation coefficient (r(2)) of 0.923.
2005 – F.PELLATI, S.BENVENUTI, M.MELEGARI – . Enantioselective LC analysis of synephrine in natural products on a protein-based chiral stationary phase. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, vol. 37, p. 839-849, ISSN: 0731-7085
Abstract – An enantioselective LC method with photodiode array detection (PAD) was developed for the enantioseparation of synephrine from C. aurantium L. var. amara fruits and phytotherapic derivatives by using a protein-based chiral stationary phase with cellobiohydrolase as the chiral selector (Chiral-CBH). Analyses were carried out on a Chiral-CBH column (100 x 4.0 mm i.d., 5 um), with a mobile phase consisting of 2-propanol (5%, w/w) in sodium phosphate buffer (pH 6.0; 10 mM) and disodium EDTA (50 uM). The flow rate was 0.8 mL/min. Detection was set at 225 nm. To identify the order of elution, the racemate was resolved by the preparation of suitable diastereoisomeric salts with antipodes of appropriate organic acids. Isolation of synephrine from C aurantium fruits and phytoproducts was performed by solid-phase extraction (SPE) with a strong cation-exchange phase. The method developed was validated and was found to be linear in the 0.40-40.14 ug/mL range (r(2) = 1.000, P < 0.0001) for both synephrine enantiomers. The limit of detection (LOD) for each enantiomer was 0.04 ug/mL. The limit of quantification (LOQ) for each enantiomer was 0.13 ug/mL. Intra-day precision (calculated as %R.S.D.) ranged from 0.03 to 0.24% for (-)-synephrine and from 0.03 to 0.35% for (+)-synephrine. Inter-day precision (calculated as %R.S.D.) ranged from 0.07 to 1.45% for (-)-synephrine and from 0.06 to 1.26% for (+)synephrine. Intra- and inter-day accuracies (calculated as %recovery) were in the ranges of 97.4-100.6 and 98.0-101.6% for (-)-synephrine, and in the ranges 97.0-101.5 and 98.1-102.8% for (+)-synephrine. The results of the application of the method to the analysis of C aurantium samples showed that (-)-synephrine was the main component. (+)-Synephrine was not detected in C aurantium fruits and was present in low concentration in the phytoproducts.
2005 – PELLATI F, BENVENUTI S, MELEGARI M .- Chromatographic performance of a new polar poly(ethyleneglycol) bonded phase for the phytochemical analysis of Hypericum perforatum L. JOURNAL OF CHROMATOGRAPHY A, vol. 1088, p. 205-217, ISSN: 0021-9673
Abstract – The aim of this study was to evaluate the chromatographic performance of a poly(ethylene glycol) (PEG) stationary phase for the HPLC analysis of the secondary metabolites (chlorogenic acid, flavonoids, phloroglucinols and naphthodianthrones) in methanolic extracts of Hypericum perforatum L. (St. John’s Wort) flowering tops, herbal medicinal products and dietary supplements. A fast and reliable method was developed. The analyses were carried out on a Supelco Discovery HS PEG column (150 mm × 4.6 mm i.d., 5 μm). A gradient mobile phase, composed of 0.1 M aqueous acetic acid solution (pH 2.8) and methanol–acetonitrile (5:4, v/v), was used. The flow rate was 1 mL/min. The photodiode array detector monitored the eluent at 270 (for chlorogenic acid, flavonoids and phloroglucinols) and 590 nm (for naphthodianthrones). The column was maintained at room temperature. The total running time was 40 min. The method was validated and showed good linearity, precision, accuracy, sensitivity and specificity. Through the above described phytochemical markers, this technique allowed the unequivocal identification and standardization of H. perforatum plant material and phytoproducts. The quantification data highlighted the fact that the products on sale, in particular those labeled as dietary supplements, varied widely in the quantitative composition of the active constituents. The developed method could be considered suitable for the quality control of H. perforatum herb and derivatives.
2005 – F. Pellati; S. Benvenuti; F. Yoshizaki; M. Melegari – Optimization and validation of HPLC methods for the analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species ( 11th Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2005) – Rimini, Italy – 25-28 Settembre 2005) ( – 11th Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2005) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Rimini ITA ) – n. volume 1 – pp. da 186 a 186 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this work, HPLC methods were developed and validated for the phytochemical analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species.
2005 – F.Pellati; S. Calò; S. Benvenuti; B. Adinolfi; A. Chicca; P. Nieri; E. Martinotti; M. Melegari – Isolation, structural elucidation and biological activity of potential antiproliferative polyacetylenes from Echinacea pallida ( Second Joint Italian-Swiss Meeting on Medicinal Chemistry (ITCHMC 2005) – Modena – 12-16 Settembre 2005) ( – Second Joint Italian-Swiss Meeting on Medicinal Chemistry (ITCHMC 2005) Abstracts ) (Divisione di Chimica Farmaceutica della Società Chimica Italiana Modena ITA ) – n. volume 1 – pp. da 112 a 112 ISBN: 0000000000 [Abstract in Atti di convegno (274)]
Abstract – In this work, the isolation, structural elucidation and biological activity of potential antiproliferative polyacetylenes and polyenes from Echinacea pallida was carried out and discussed.
2005 – R.Bruni; F. Pellati; MG. Bellardi; S. Benvenuti; S. Paltrinieri; A. Bertaccini; A. Bianchi – Herbal drug quality and phytochemical composition of Hypericum perforatum L. affected by ash yellows phytoplasma infection – JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY – n. volume 53 – pp. da 964 a 968 ISSN: 0021-8561 [Articolo in rivista (2]
Abstract – Qualitative/quantitative phytochemical variations were observed in dried flowering tops of cultivated Hypericum perforatum L. cv. Zorzi infected by phytoplasmas of the ash yellows class, identified by direct and nested polymerase chain reaction (PCR); this is the first report of ribosomial group 16SrVII phytoplasmas in St. John’s Wort. Methanolic extracts of healthy and infected plants were separated by reversed phase high-performance liquid chromatography to quantify naphthodianthrones and flavonoids, while essential oils were analyzed by means of gas chromatography (GC)-GC/MS. The affected plants exhibited decreased amounts of rutin (1.96 +/- 0.23 vs 4.96 +/- 0.02 mg/g), hyperoside (2.38 +/- 0.21 vs 3.04 +/- 0.05 mg/g), isoquercitrin (1.47 +/- 0.04 vs 3.50 +/- 0.08 mg/g), amentoflavone (0.12 +/- 0.01 vs 0.39 +/- 0.02 mg/g), and pseudohypericin (1.41 +/- 0.23 vs 2.29 +/- 0.07 mg/g), whereas the chlorogenic acid content was doubled (1.56 +/- 0.11 vs 0.77 +/- 0.02 mg/g). Hypericin, quercitrin, and quercetin contents were not severely affected. The essential oil yield was drastically reduced in infected material (0.11 vs 0.75% in healthy material) and revealed an increased abundance of sesquiterpenes (beta-caryophyllene, delta-elemene, and germacrene D, in particular) and a matching decrease in monoterpene hydrocarbons and aliphatics. The consequences that the phytopathological condition of cultivated H. perforatum plants has on the commercial quality, market value, and, therapeutic efficacy are outlined.
2005 – F.Pellati; S. Benvenuti; M. Melegari – Enantioselective LC analysis of synephrine in natural products on a protein-based chiral stationary phase – JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL
2005 -F. PELLATI; S. BENVENUTI; F. YOSHIZAKI; D. BERTELLI; M.C. ROSSI – Headspace solid-phase microextraction-gas chromatography-mass spectrometry analysis of the volatile compounds of Evodia species fruits – JOURNAL OF CHROMATOGRAPHY A – n. volume 1087 – pp. da 265 a 273 ISSN: 0021-9673 [Articolo in rivista (262)]
Abstract – In this study the investigation of the aroma compounds of dried fruits of Evodia rutaecarpa (Juss.) Benth. and E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (i.e. E. officinalis Dode) (Rutaceae family) was carried out to identify the odorous target components responsible for the characteristic aroma of these valuable natural products. To avoid the traditional and more time-consuming hydrodistillation, the analyses were carried out by means of headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography–mass spectrometry (GC–MS). The SPME headspace volatiles were collected using a divinylbenzene–carboxen–polydimethylsiloxane (DVB–CAR–PDMS) fiber. The extraction conditions were optimized using a response surface experimental design to analyze the effect of three factors: extraction temperature, equilibrium time and extraction time. The best response was obtained when the extraction temperature was around 80 °C, equilibrium time near 25 min and extraction time close to 18 min. Analyses were performed by GC–MS with a 5% diphenyl–95% dimethyl polysiloxane (30 m × 0.25 mm I.D., film thickness 0.25 μm) capillary column using He as the carrier gas and a programmed temperature run. The main components of the HS-SPME samples of E. rutaecarpa (concentration >3.0%) were limonene (33.79%), β-elemene (10.78%), linalool (8.15%), myrcene (5.83%), valencene (4.73%), β-caryophyllene (4.62%), linalyl acetate (4.13%) and α-terpineol (3.99%). As for E. officinalis, the major compounds were myrcene (32.79%), limonene (18.36%), β-caryophyllene (9.92%), trans-β-ocimene (6.04%), linalool (5.88%), β-elemene (7.85%) and valencene (4.62%).
2006 – F.PELLATI, S.BENVENUTI, F. YOSHIZAKY, M.MELEGARI . Development and validation of HPLC methods for the analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species. JOURNAL OF SEPARATION SCIENCE, vol. 29, p. 641-649, ISSN: 1615-9306
Abstract – The aim of this study was to evaluate the chromatographic performance of a PEG stationary phase, in comparison with those of C-18 columns, for the HPLC analysis of phenethylamine ((+/-)-synephrine) and indoloquinazoline (rutaecarpine and evodiamine) alkaloids in methanolic extracts of fruits of Evodia rutaecarpa (Juss.) Benth. and E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (i.e., E. officinalis Dode) (Rutaceae family). The method was validated and showed good linearity, precision, accuracy, sensitivity, and specificity. The highest content of both phenethylamine and indoloquinazoline alkaloids was found in methanolic fruit extracts of E. rutaecarpa, and it was closely related to the degree of maturity. E. officinalis fruits displayed low amounts of both types of alkaloids. Furthermore, an enantioselective HPLC method for the enantioseparation of (+/-)-synephrine from Evodia fruits was applied, by using a protein-based chiral stationary phase with cellobiohydrolase (CBH) as the chiral selector (Chiral-CBH). Isolation of synephrine from Evodia aqueous fruit extracts was carried out by strong cation-exchange SPE. The results of the application of the method to the analysis of Evodia samples showed that (-)-synephrine was the main component while (+)-synephrine was present in low concentration.
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2006 – F. Pellati; S. Calo; S. Benvenuti; B. Adinolfi; P. Nieri; M. Melegari – Isolation and structure elucidation of cytotoxic polyacetylenes and polyenes from Echinacea pallida – PHYTOCHEMISTRY – n. volume 67 – pp. da 1359 a 1364 ISSN: 0031-9422 [Articolo in rivista (262) – Articolo su rivista]
Abstract – Bioassay-guided fractionation of n-hexane extracts of Echinacea pallida (Asteraceae) roots led to the isolation and structure elucidation of two polyacetylenes (1, 3) and three polyenes (2, 4, 5). Two are known hydroxylated compounds, namely 8-hydroxy-pentadeca-(9E)-ene-11,13-diyn-2-one (1) and 8-hydroxy-pentadeca-(9E,13Z)-dien-11-yn-2-one (2). Two dicarbonylic constituents, namely pentadeca-(9E)-ene-11,13-diyne-2,8-dione (3) and pentadeca-(9E,13Z)-dien-11-yne-2,8-dione (4), were isolated and characterized for the first time. Furthermore, the structure elucidation of pentad eca-(8Z,13Z)-dien-11-yn-2-one (5) is described. The structure of the compounds isolated was determined on the basis of UV, IR, NMR (including 1D and 2D NMR experiments, such as H-1-H-1 gCOSY, gHSQC-DEPT, gHMBC, gNOESY) and MS spectroscopic data. The cytotoxic activity of the isolated constituents against MIA PaCa-2 human pancreatic adenocarcinoma cells was evaluated in the concentration range 1-100 mu g/ml. Results show that the hydroxylated compounds (1, 2) have low cytotoxicity, while the more hydrophobic polyacetylenes (3) and polyenes (4, 5) displayed moderate activity.
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2006 – F. Pellati; S. Calo; S. Benvenuti; B. Adinolfi; P. Nieri; M. Melegari – Isolation and structure elucidation of cytotoxic polyacetylenes and polyenes from Echinacea pallida – PHYTOCHEMISTRY – n. volume 67 – pp. da 1359 a 1364 ISSN: 0031-9422 [Articolo in rivista (262) – Articolo su rivista]
Abstract – Bioassay-guided fractionation of n-hexane extracts of Echinacea pallida (Asteraceae) roots led to the isolation and structure elucidation of two polyacetylenes (1, 3) and three polyenes (2, 4, 5). Two are known hydroxylated compounds, namely 8-hydroxy-pentadeca-(9E)-ene-11,13-diyn-2-one (1) and 8-hydroxy-pentadeca-(9E,13Z)-dien-11-yn-2-one (2). Two dicarbonylic constituents, namely pentadeca-(9E)-ene-11,13-diyne-2,8-dione (3) and pentadeca-(9E,13Z)-dien-11-yne-2,8-dione (4), were isolated and characterized for the first time. Furthermore, the structure elucidation of pentad eca-(8Z,13Z)-dien-11-yn-2-one (5) is described. The structure of the compounds isolated was determined on the basis of UV, IR, NMR (including 1D and 2D NMR experiments, such as H-1-H-1 gCOSY, gHSQC-DEPT, gHMBC, gNOESY) and MS spectroscopic data. The cytotoxic activity of the isolated constituents against MIA PaCa-2 human pancreatic adenocarcinoma cells was evaluated in the concentration range 1-100 mu g/ml. Results show that the hydroxylated compounds (1, 2) have low cytotoxicity, while the more hydrophobic polyacetylenes (3) and polyenes (4, 5) displayed moderate activity.
2006 F. Pellati; S. Benvenuti; F. Yoshizaki; M. Melegari ( 2006 ) – Development and validation of )HPLC methods for the analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species – JOURNAL OF SEPARATION SCIENCE – n. volume 29 – pp. da 641 a 649 ISSN: 1615-9306 [Articolo in rivista (262) – Articolo su rivista]
Abstract – The aim of this study was to evaluate the chromatographic performance of a PEG stationary phase, in comparison with those of C-18 columns, for the HPLC analysis of phenethylamine ((+/-)-synephrine) and indoloquinazoline (rutaecarpine and evodiamine) alkaloids in methanolic extracts of fruits of Evodia rutaecarpa (Juss.) Benth. and E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (i.e., E. officinalis Dode) (Rutaceae family). The method was validated and showed good linearity, precision, accuracy, sensitivity, and specificity. The highest content of both phenethylamine and indoloquinazoline alkaloids was found in methanolic fruit extracts of E. rutaecarpa, and it was closely related to the degree of maturity. E. officinalis fruits displayed low amounts of both types of alkaloids. Furthermore, an enantioselective HPLC method for the enantioseparation of (+/-)-synephrine from Evodia fruits was applied, by using a protein-based chiral stationary phase with cellobiohydrolase (CBH) as the chiral selector (Chiral-CBH). Isolation of synephrine from Evodia aqueous fruit extracts was carried out by strong cation-exchange SPE. The results of the application of the method to the analysis of Evodia samples showed that (-)-synephrine was the main component while (+)-synephrine was present in low concentration.
2007 -F. Pellati; S. Benvenuti; M. Melegari – Determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase ( 12th Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2007) – Isola d’Elba, Italy – 23-26 Settembre 2007) ( – Recent Developments in Pharmaceutical Analysis 12th International Meeting (RDPA 2007) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Firenze ITA ) – n. volume 1 – pp. da 230 a 230 ISBN: 0000000000 [Abstract in Atti di convegno (274) – Abstract in Atti di Convegno]
Abstract – In this research, the determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase was carried out. Version:1.0 StartHTML:0000000167 EndHTML:0000002592 StartFragment:0000000481 EndFragment:0000002576
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2007 F. Pellati; S. Benvenuti; M. Melegari – Determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase ( 12th Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2007) – Isola d’Elba, Italy – 23-26 Settembre 2007) ( – Recent Developments in Pharmaceutical Analysis 12th International Meeting (RDPA 2007) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Firenze ITA ) – n. volume 1 – pp. da 230 a 230 ISBN: 0000000000 [Abstract in Atti di convegno (274) – Abstract in Atti di Convegno]
Abstract In this research, the determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase was carried out.
2007 F.PELLATI; S. CALÒ; S. BENVENUTI – High-performance liquid chromatography analysis of polyacetylenes and polyenes in Echinacea pallida by using a monolithic reversed-phase silica column – JOURNAL OF CHROMATOGRAPHY A – n. volume 1149 – pp. da 56 a 65 ISSN: 0021-9673 [Articolo in rivista (262) – Articolo su rivista]
AbstractIn this study, a RP-HPLC method for the analysis of polyacetylenes and polyenes in Echinacea pallida roots and phytopharmaceuticals was developed. The reference compounds used for quantification were isolated from the plant material and their structures were determined on the basis of the analysis of UV, IR, NMR and MS data. The complete structure elucidation of three compounds, namely 8-hydroxy-tetradec-(9E)-ene-11,13-diyn-2-one (1), tetradec-(8Z)-ene-11,13-diyn-2-one (6) and pentadec-(8Z)-en-2-one (9) is described. In the analysis of the n-hexane extracts of E. pallida roots, the comparison between conventional and monolithic columns showed that the elution order in both cases is identical and the selectivity is equivalent. However, the retention times achieved by the monolithic column are shorter, resulting in a faster separation (20 min).Therefore, the analyses were carried out on a Chromolith Performance RP-18e (100 mm× 4.6 mm i.d.), with a gradient mobile phase composed by H2O and ACN at the flow rate of 2 mL/min. The column was thermostatted at 20 ◦C. The photodiode array detector monitored the eluent at 210 nm. The validation procedure confirmed that this technique affords reliable analysis of these components and is appropriate for the quality control of complex matrices, such as E. pallida roots and phytopharmaceuticals.
2007 A. Chicca; B. Adinolfi; E. Martinotti; S. Fogli; M.C. Breschi; F. Pellati; S. Benvenuti; P. Nieri- Cytotoxic effects of Echinacea root hexanic extracts on human cancer cell lines – JOURNAL OF ETHNOPHARMACOLOGY – n. volume 110 – pp. da 148 a 153 ISSN: 0378-8741 [Articolo in rivista (262) –
Abstract Echinacea is one of the most widely used alternative medicine in the world. Intake of Echinacea preparations is common among patients with advanced malignancies enrolled onto phase I chemotherapy trials; however, to our knowledge, no data are available regarding the possible direct effect of Echinacea species on human cancer cells. The purpose of the present study was to investigate potential in vitro cytotoxic and pro-apoptotic properties of hexanic root extract of the three medicinal Echinacea (Asteraceae) species (Echinacea pallida (Nutt.) Nutt., Echinacea angustifolia DC. var. angustifolia, Echinacea purpurea (L.) Moench.) on the human pancreatic cancer MIA PaCa-2 and colon cancer COLO320 cell lines. We demonstrated, for the first time, that all the three species reduced cell viability in a concentration- and time-dependent manner; Echinacea pallida was the most active species with IC(50)s of 46.41 +/- 0.87 and 10.55 +/- 0.70 ug/ml in MIA PaCa-2 and COLO320 cells, respectively. Echinacea pallida extract was able to induce apoptosis by increasing significantly caspase 3/7 activity and promoting nuclear DNA fragmentation. These results represent the starting point to establish viable scientific evidence on the possible role of Echinacea species in medical oncology.
2007 – F. PELLATI; S. BENVENUTI – Fast high-performance liquid chromatography analysis of phenethylamine alkaloids in Citrus natural products on a pentafluorophenylpropyl stationary phase – JOURNAL OF CHROMATOGRAPHY A – n. volume 1165 – pp. da 58 a 66 ISSN: 0021-9673 [Articolo in rivista (262) – Articolo su rivista]
Abstract – In this study, the chromatographic performance of a pentafluorophenylpropyl (PFPP) stationary phase was evaluated for the rapid separation of phenethylamine alkaloids (i.e. (±)-octopamine, (±)-synephrine, tyramine, N-methyltyramine and hordenine) in Citrus aurantium plant material (fruits and peel), various Citrus species, extracts and dietary supplements claiming to contain C. aurantium. The problems of phenethylaminealkaloid separation, such as peak tailing, low retention and low resolution, were successfully solved with this stationary phase. The parameters used for the method optimization included the mobile phase counter ion concentration and column temperature. A Discovery HS F5 column (150mm×4.6mm i.d., 5m) was used, with an isocratic mobile phase composed of 10mM ammonium acetate in 90:10 ACN–H2O (v/v), at a flow rate of 1.0 mL/min. The column temperature was set at 20 ◦C. The photodiode array detector monitored the eluent at 225 nm. The total analysis time was 10 min. The validation parameters, such as linearity, sensitivity, accuracy, precision and specificity, were found to be highly satisfactory. With a simple sample preparation procedure, different matrices were successfully analyzed for their alkaloid content. The results indicated that the products on sale, labeled as dietary supplements, vary widely in the quantitative composition of the active constituents: the amount of (±)-synephrine, the major alkaloid, in such products ranged from 0.65 to 27.41 mg/g. The other compounds were either not detected or were present at low levels. The developed method can be considered suitable for the quality control of Citrus plant material and commercial products.
2008 – F. Pellati; S. Benvenuti – Variability of the active compounds in phytotherapic products with antidepressant activity: the case of Hypericum perforatum L. ( Therapeutic and toxic effects of central nervous system drugs: the importance of drug monitoring – Bologna, Italy – 2-3 Ottobre 2008) ( – Therapeutic and toxic effects of central nervous system drugs: the importance of drug monitoring ) (Alma Mater Studiorum, Università di Bologna Bologna ITA ) – n. volume 1 – pp. da 72 a 73 ISBN: 0000000000 [Contributo in Atti di convegno (273) – Relazione in Atti di Convegno]
Abstract – In this presentation, the variability of the active compounds in phytotherapic products containing Hypericum perforatum L. was discussed.
2008 S. Morandi; F. Pellati; S. Benvenuti; F. Prati – Total synthesis of a dienynone from Echinacea pallida – TETRAHEDRON – n. volume 64 – pp. da 6324 a 6328 ISSN: 0040-4020 [Articolo in rivista (262) –
Abstract The first total synthesis of (8Z,13Z)-pentadeca-8,13-dien-11-yn-2-one is described. This dienynone was recently isolated from the n-hexane extract of Echinacea pallida roots and displayed a selective cytotoxic activity toward cancer cells, thus featuring as a potential anticancer lead. The product was obtained in 11 steps in 25% overall yield.
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2008 – N. Romiti; F. Pellati; P. Nieri; S. Benvenuti; B. Adinolfi; E. Chieli. ( 2008 ) – P-Glycoprotein inhibitory activity of lipophilic constituents of Echinacea pallida roots in a human proximal tubular cell line – PLANTA MEDICA – n. volume 74 – pp. da 264 a 266 ISSN: 0032-0943 [Articolo in rivista (262)
Abstract The n-hexane root extracts from Echinacea pallida, Echinacea angustifolia and Echinacea purpurea were evaluated for inhibitionof the multidrug transporter P-glycoprotein (Pgp) activity,the product of the ABCB1 gene, involved in cancer multidrug resistance (MDR) and in herb-drug or drug-drug interactions. Thebiological assay was performed using the human proximal tubule HK-2 cell line that constitutively expresses ABCB1. The n-hexaneextracts of all three species reduced the efflux of the Pgp probe calcein-AM from HK-2 cells two-fold in a concentration-dependent manner, and E. pallida was found to be the most active species. For the first time, two polyacetylenes and three polyenes, isolated from the n-hexane extract of E. pallida roots by a bioassay- guided fractionation, were found to be able to reduce Pgp activity. Pentadeca-(8Z,13Z)-dien-11-yn-2-one was the most efficient compound, being able to decrease the calcein-AM efflux about three-fold with respect to the control at 30 μg/m
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2008 F. Pellati; S. Benvenuti – Determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase – JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS – n. volume 48 – pp. da 254 a 263 ISSN: 0731-7085 [Articolo in rivista (262)
Abstract In this study a pentafluorophenylpropyl (PFPP) stationary phase was applied to the fast and reliable qualitative and quantitative analysis of ephedrine alkaloids in Ephedra plant material and derivatives. A Discovery HS F5 column (150 mm × 4.6 mm i.d., 5 um) was used, with an isocratic mobile phase composed of ammonium acetate (7 mM) in acetonitrile–water (90:10, v/v), at a flow rate of 1.0 ml/min. The column temperature was set at 45 °C. UV detection was set at 215 and 225 nm. The total analysis time was 16 min. The validation parameters, such as linearity, sensitivity, accuracy, precision and specificity, were found to be highly satisfactory. Sonication and microwave extractions were compared in order to optimize the yield of the target analytes. Under the optimized extraction conditions (based on two cycles of sonication with methanol at 40 °C for 15 min), different matrices containing Ephedra were successfully analyzed for their alkaloid content. The method was applied to the analysis of standard reference materials (SRMs) containing Ephedra. Furthermore, the developed technique allowed the simultaneous determinationof ephedrine alkaloids and synephrine, the main phenethylamine alkaloid of Citrus aurantium, that has replaced Ephedra in dietary supplements used in the treatment of obesity. The results indicated that this procedure is suitable for the phytochemical analysis of Ephedra plant material and extracts, and can be applied to demonstrate the label claims for product content, including the absence of ephedrine alkaloids in Ephedra-freeproducts.
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2008 A. Chicca; F. Pellati; B. Adinolfi; A. Matthias; I. Massarelli; S. Benvenuti; E. Martinotti; A.M. Bianucci; K. Bone; R. Lehmann; P. Nieri ( 2008 ) – Cytotoxic activity of polyacetylenes and polyenes isolated from roots of Echinacea pallida – BRITISH JOURNAL OF PHARMACOLOGY – n. volume 153 – pp. da 879 a 885 ISSN: 0007-1188 [Articolo in rivista
Abstract – BACKGROUND AND PURPOSE: The n-hexane extracts of the roots of three medicinally used Echinacea species exhibited cytotoxic activity on human cancer cell lines, with Echinacea pallida found to be the most cytotoxic. Acetylenes are present in E. pallida lipophilic extracts but essentially absent in extracts from the other two species. In the present study, the cytotoxic effects of five compounds, two polyacetylenes (namely, 8-hydroxy-pentadeca-(9E)-ene-11,13-diyn-2-one (1) and pentadeca-(9E)-ene-11,13-diyne-2,8-dione (3)) and three polyenes (namely, 8-hydroxy-pentadeca-(9E,13Z)-dien-11-yn-2-one (2), pentadeca-(9E,13Z)-dien-11-yne-2,8-dione (4) and pentadeca-(8Z,13Z)-dien-11-yn-2-one (5)), isolated from the n-hexane extract of E. pallida roots by bioassay-guided fractionation, were investigated and the potential bioavailability of these compounds in the extract was studied. EXPERIMENTAL APPROACH: Cytotoxic effects were assessed on human pancreatic MIA PaCa-2 and colonic COLO320 cancer cell lines. Cell viability was evaluated by the WST-1 assay and apoptotic cell death by the cytosolic internucleosomal DNA enrichment and the caspase 3/7 activity tests. Caco-2 cell monolayers were used to assess the potential bioavailability of the acetylenes. KEY RESULTS: The five compounds exhibited concentration-dependent cytotoxicity in both cell types, with a greater potency in the colonic cancer cells. Apoptotic cell death was found to be involved in the cytotoxic effect of the most active, compound 5. Compounds 2 and 5 were found to cross the Caco-2 monolayer with apparent permeabilities above 10 x 10(-6) cm s(-1). CONCLUSIONS AND IMPLICATIONS: Compounds isolated from n-hexane extracts of E. pallida roots have a direct cytotoxicity on cancer cells and good potential for absorption in humans when taken orally.
2008 – S. Morandi; F. Pellati; S. Benvenuti; F. Prati – Total synthesis of a dienynone from Echinacea pallida – TETRAHEDRON – n. volume 64 – pp. da 6324 a 6328 ISSN: 0040-4020 Articolo in rivista
Abstract – The first total synthesis of (8Z,13Z)-pentadeca-8,13-dien-11-yn-2-one is described. This dienynone was recently isolated from the n-hexane extract of Echinacea pallida roots and displayed a selective cytotoxic activity toward cancer cells, thus featuring as a potential anticancer lead. The product was obtained in 11 steps in 25% overall yield.
2008 – N. Romiti; F. Pellati; P. Nieri; S. Benvenuti; B. Adinolfi; E. Chieli. – P-Glycoprotein inhibitory activity of lipophilic constituents of Echinacea pallida roots in a human proximal tubular cell line – PLANTA MEDICA – n. volume 74 – pp. da 264 a 266 ISSN: 0032-0943 [Articolo in rivista]
Abstract – The n-hexane root extracts from Echinacea pallida, Echinacea angustifolia and Echinacea purpurea were evaluated for inhibitionof the multidrug transporter P-glycoprotein (Pgp) activity,the product of the ABCB1 gene, involved in cancer multidrug resistance (MDR) and in herb-drug or drug-drug interactions. Thebiological assay was performed using the human proximal tubule HK-2 cell line that constitutively expresses ABCB1. The n-hexaneextracts of all three species reduced the efflux of the Pgp probe calcein-AM from HK-2 cells two-fold in a concentration-dependent manner, and E. pallida was found to be the most active species. For the first time, two polyacetylenes and three polyenes, isolated from the n-hexane extract of E. pallida roots by a bioassay- guided fractionation, were found to be able to reduce Pgp activity. Pentadeca-(8Z,13Z)-dien-11-yn-2-one was the most efficient compound, being able to decrease the calcein-AM efflux about three-fold with respect to the control at 30 μg/mL.
2008 – S. Morandi; F. Pellati; C. Ori; B. Adinolfi; P. Nieri; S. Benvenuti; F. Prati – Isolation, structure elucidation and total synthesis of a cytotoxic dienone from Echinacea pallida – ORGANIC & BIOMOLECULAR CHEMISTRY – n. volume 6 – pp. da 4333 a 4339 ISSN: 1477-0520 [Articolo su rivista]
Abstract – The isolation and structure characterization of a dienone from the roots of Echinacea pallida, namely (8Z,11Z)-pentadeca-8,11-dien-2-one, are described here. To assess the configuration of this secondary metabolite, the stereoselective total synthesis of the two isomeric forms, 8Z,11Z)- and (8Z,11E)-pentadeca-8,11-dien-2-one, was undertaken and the structure elucidation of the naturalcompound was unambiguously carried out. The cytotoxic activity of both isomers was also evaluated on a human T cell leukaemia cancer line (Jurkat cells). The results indicated that these compounds exert a dose-dependent cytotoxicity with a medium-level potency on the tested cell line.
2008 – F. Pellati; S. Benvenuti – Determination of ephedrine alkaloids in Ephedra natural products using HPLC on a pentafluorophenylpropyl stationary phase – JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS – n. volume 48 – pp. da 254 a 263 ISSN: 0731-7085 [Articolo in rivista]
Abstract – In this study a pentafluorophenylpropyl (PFPP) stationary phase was applied to the fast and reliable qualitative and quantitative analysis of ephedrine alkaloids in Ephedra plant material and derivatives. A Discovery HS F5 column (150 mm × 4.6 mm i.d., 5 um) was used, with an isocratic mobile phase composed of ammonium acetate (7 mM) in acetonitrile–water (90:10, v/v), at a flow rate of 1.0 ml/min. The column temperature was set at 45 °C. UV detection was set at 215 and 225 nm. The total analysis time was 16 min. The validation parameters, such as linearity, sensitivity, accuracy, precision and specificity, were found to be highly satisfactory. Sonication and microwave extractions were compared in order to optimize the yield of the target analytes. Under the optimized extraction conditions (based on two cycles of sonication with methanol at 40 °C for 15 min), different matrices containing Ephedra were successfully analyzed for their alkaloid content. The method was applied to the analysis of standard reference materials (SRMs) containing Ephedra. Furthermore, the developed technique allowed the simultaneous determinationof ephedrine alkaloids and synephrine, the main phenethylamine alkaloid of Citrus aurantium, that has replaced Ephedra in dietary supplements used in the treatment of obesity. The results indicated that this procedure is suitable for the phytochemical analysis of Ephedra plant material and extracts, and can be applied to demonstrate the label claims for product content, including the absence of ephedrine alkaloids in Ephedra-freeproducts.
2008 – A. Chicca; F. Pellati; B. Adinolfi; A. Matthias; I. Massarelli; S. Benvenuti; E. Martinotti; A.M. Bianucci; K. Bone; R. Lehmann; P. Nieri – Cytotoxic activity of polyacetylenes and polyenes isolated from roots of Echinacea pallida – BRITISH JOURNAL OF PHARMACOLOGY – n. volume 153 – pp. da 879 a 885 ISSN: 0007-1188 [Articolo in rivista]
Abstract – BACKGROUND AND PURPOSE: The n-hexane extracts of the roots of three medicinally used Echinacea species exhibited cytotoxic activity on human cancer cell lines, with Echinacea pallida found to be the most cytotoxic. Acetylenes are present in E. pallida lipophilic extracts but essentially absent in extracts from the other two species. In the present study, the cytotoxic effects of five compounds, two polyacetylenes (namely, 8-hydroxy-pentadeca-(9E)-ene-11,13-diyn-2-one (1) and pentadeca-(9E)-ene-11,13-diyne-2,8-dione (3)) and three polyenes (namely, 8-hydroxy-pentadeca-(9E,13Z)-dien-11-yn-2-one (2), pentadeca-(9E,13Z)-dien-11-yne-2,8-dione (4) and pentadeca-(8Z,13Z)-dien-11-yn-2-one (5)), isolated from the n-hexane extract of E. pallida roots by bioassay-guided fractionation, were investigated and the potential bioavailability of these compounds in the extract was studied. EXPERIMENTAL APPROACH: Cytotoxic effects were assessed on human pancreatic MIA PaCa-2 and colonic COLO320 cancer cell lines. Cell viability was evaluated by the WST-1 assay and apoptotic cell death by the cytosolic internucleosomal DNA enrichment and the caspase 3/7 activity tests. Caco-2 cell monolayers were used to assess the potential bioavailability of the acetylenes. KEY RESULTS: The five compounds exhibited concentration-dependent cytotoxicity in both cell types, with a greater potency in the colonic cancer cells. Apoptotic cell death was found to be involved in the cytotoxic effect of the most active, compound 5. Compounds 2 and 5 were found to cross the Caco-2 monolayer with apparent permeabilities above 10 x 10(-6) cm s(-1). CONCLUSIONS AND IMPLICATIONS: Compounds isolated from n-hexane extracts of E. pallida roots have a direct cytotoxicity on cancer cells and good potential for absorption in humans when taken orally.
2009 – F. Pellati; G. Cannazza; G. Orlandini; S. Benvenuti – Study on the enantiomerization of synephrine by chiral high-performance liquid chromatography ( Recent Developments in Pharmaceutical Analysis 13th International Meeting (RDPA 2009) – Milano – 9-12 Settembre 2009) ( – Recent Developments in Pharmaceutical Analysis 13th International Meeting (RDPA 2009) Abstract Book ) (Gruppo Intradivisionale di Analisi Farmaceutica della Divisione di Chimica Farmaceutica della Società Chimica Italiana Milano ITA ) – n. volume 1 – pp. da 100 a 100 ISBN: 0000000000 [Abstract in Atti di convegno]
2009 – G. Orlandini; F. Pellati; S. Benvenuti – Stability study on a cytotoxic dienynone isolated from Echinacea pallida ( IX Giornata della Chimica dell’Emilia-Romagna – Bologna – 4 Dicembre 2009) ( – IX Giornata della Chimica dell’Emilia-Romagna ) (Società Chimica Italiana Bologna ITA ) – n. volume * – pp. da 64 a 64 ISBN: 0000000000 [Abstract in Atti di convegno]
Abstract – Recent studies have demonstrated that some of the secondary metabolites (polyacetylenes and polyenes) isolated from the raw lipophilic extract of Echinacea pallida roots have good cytotoxic activity on cancer cell lines3. In particular, pentadeca-(8Z,13Z)-dien-11-yn-2-one was found to be the most active constituent, with an IC50 value of 32.17 ± 3.98 uM on pancreatic MIA PaCa-2 cancer cells and 2.34 ± 0.33 uM on colonic COLO320 cancer cells, after 72 h of exposure. This compound is present in the roots in a not-oxidized form but, during the drying process and the subsequent storage, a change in its concentration was observed, which has been attributed to a possible oxidation process. As part of our ongoing research on bioactive metabolites from Echinacea, the aim of this research was the evaluation of the stability of this secondary metabolite by RP-HPLC-DAD2 under the experimental conditions applied during the biological assays. The study was initially carried out on the lipophilic crude extract obtained from E. pallida roots and then it was focused on the individual component isolated and purified from the plant material by preparative chromatographic techniques. The results showed a moderate change in the HPLC profiles of E. pallida crude extract in 72 h, with an increase of hydroxylated derivatives. The oxidation kinetic of the most active compound, pentadeca-(8Z,13Z)-dien-11-yn-2-one, was found to be quite slow, since the % area of the intermediate hydroperoxide was about 25% after 72 h of exposure. The final product of the oxidation process, i.e. the hydroxilated derivative (i.e. 8-hydroxy-pentadeca-(9E,13Z)-dien-11-yn-2-one, available thanks to a synthetic approach), was not observed in the HPLC chromatogram after 72 h. Considering that during the first 24 h of exposure, the % area of the hydroperoxide intermediate is below 15%, the observed cytotoxic activity can be mainly attributed to the genuine not oxidized compound.
2009 – F. Pellati; S. Benvenuti – Recent advances in the chromatographic analysis of phenethylamine alkaloids in natural products ( Recent Developments in Pharmaceutical Analysis 13th International Meeting (RDPA 2009) – Milano, Italy – September 9-12, 2009) ( – RecentF. Pellati; R. Bruni; M.G. Bellardi; A. Bertaccini; S. Benvenuti ( 2009 ) – Optimization and validation of a high-performance liquid chromatography method for the analysis of cardiac glycosides in Digitalis lanata – JOURNAL OF CHROMATOGRAPHY A – n. volume 1216 – pp. da 3260 a 3269 ISSN: 0021-9673 [Articolo in rivista]
Abstract – In this study, a simple and reliable HPLC method for the qualitative and quantitative analysis of cardiac glycosides in Digitalis lanata Ehrh. raw material was developed and applied to healthy and phytoplasma-infected plants. The target analytes cover a broad range of secondary metabolites, including primary, secondary and tertiary glycosides and the corresponding aglycones. The sample preparation was carried out by sonication of the plant material with 70% (v/v) aqueous methanol at room temperature, followed by reversed-phase solid-phase extraction purification from interfering pigments. The HPLC analyses were performed on a Symmetry C(18) column (75 mm x 4.6mm I.D., 3.5 microm), with a gradient elution composed of water and acetonitrile, at a flow rate of 1.0 mL/min. The column temperature was set at 20 degrees C and the photodiode array detector monitored the eluent at 220 nm. The method was validated with respect to ICH guidelines and the validation parameters were found to be highly satisfactory. The application of the method to the analysis of D. lanata leaves indicated that air-drying was the optimum method for raw material processing when compared with freeze-drying. The analysis of healthy and phytoplasma-infected plants demonstrated that the secondary metabolite mainly affected by the pathogen presence was lanatoside C (153.2 ug/100 mg versus 76.1 ug/100 mg). Considering the importance of D. lanata plant material as source of cardiac glycosides, the developed method can be considered suitable for the phytochemical analysis and for the quality assurance of D. lanata used for pharmaceutical purpose.
2009 – F. Pellati; R. Bruni; M.G. Bellardi; A. Bertaccini; S. Benvenuti – Optimization and validation of a high-performance liquid chromatography method for the analysis of cardiac glycosides in Digitalis lanata – JOURNAL OF CHROMATOGRAPHY A – n. volume 1216 – pp. da 3260 a 3269 ISSN: 0021-9673 [Articolo in rivista]
Abstract – In this study, a simple and reliable HPLC method for the qualitative and quantitative analysis of cardiac glycosides in Digitalis lanata Ehrh. raw material was developed and applied to healthy and phytoplasma-infected plants. The target analytes cover a broad range of secondary metabolites, including primary, secondary and tertiary glycosides and the corresponding aglycones. The sample preparation was carried out by sonication of the plant material with 70% (v/v) aqueous methanol at room temperature, followed by reversed-phase solid-phase extraction purification from interfering pigments. The HPLC analyses were performed on a Symmetry C(18) column (75 mm x 4.6mm I.D., 3.5 microm), with a gradient elution composed of water and acetonitrile, at a flow rate of 1.0 mL/min. The column temperature was set at 20 degrees C and the photodiode array detector monitored the eluent at 220 nm. The method was validated with respect to ICH guidelines and the validation parameters were found to be highly satisfactory. The application of the method to the analysis of D. lanata leaves indicated that air-drying was the optimum method for raw material processing when compared with freeze-drying. The analysis of healthy and phytoplasma-infected plants demonstrated that the secondary metabolite mainly affected by the pathogen presence was lanatoside C (153.2 ug/100 mg versus 76.1 ug/100 mg). Considering the importance of D. lanata plant material as source of cardiac glycosides, the developed method can be considered suitable for the phytochemical analysis and for the quality assurance of D. lanata used for pharmaceutical purpose
Presentazioni P.P. (M.M.): illustrano gli aspetti fitochimici delle droghe vegetali e dei derivati fitoterapici, e protocolli e criteri per i controlli di qualità
2008 Università di Modena e Reggio E.