Laura Salvini

Tocopherol, fatty acid and phytosterol content in seeds of nine wild taxa of Sicilian Brassica (Cruciferae)

Abstract The purpose of this study was to determine tocopherol (Toc), fatty acid and phytosterol content in dry seeds of nine wild taxa of Sicilian Brassica sect. Brassica. Analyses were performed by gas chromatography (GC)‐mass spectroscopy (MS). The biochemical analysis of dry seeds showed that in the nine taxa examined, α‐ and γ‐Toc were the major isomers, δ‐Toc was present in traces and β‐Toc was never found. All taxa had a similar fatty acid composition with C 16:0 (357.2 mg kg−1 DW), C 18:1 (393.94 mg kg−1 DW) and C 22:1 (618.29 mg kg−1 DW) acids present in higher amount. Unsaturated fatty acids (52.5–66.5%) were more abundant than saturated (33.5–47.5%) ones relative to the total fatty acids. Moreover, sitosterol was the most abundant (61.7–69.6%) of all phytosterols in all species and subspecies, followed by brassicasterol and campesterol at 16.0–19.3% and 11.9–20.7%, respectively. Total Toc content and the relative percentage of single isoforms were useful to discriminate seeds of B. incana, B. macrocarpa, B. rupestris group and B. villosa group. On the basis of these results, B. villosa subsp. villosa, B. villosa subsp. bivoniana, B. villosa subsp. drepanensis and B. villosa subsp. tinei can be ascribed to the B. villosa group. These taxa, having the highest T‐Toc, γ‐Toc and phytosterol contents, are potentially the most bioactive ones, and could, therefore, be exploited in breeding programmes to develop genotypes with putative positive effects on human health. Abbreviations: B, Brassica; DW, dry weight; FA, fatty acids; RH, relative humidity; T‐FA, total fatty acids; T‐Phy, total phytosterols; T‐Toc, total tocopherol; Toc, tocopherol

Synthetic Approach, Regio- and Stereochemical Characterization and Differentiation of New Potential Antioxidant C- And O-Arylglycosides

Two series of C- and O-arylglycosides with potential antioxidant properties have been synthesized, characterized and structurally differentiated. Reinvestigation of a synthetic approach has provided better insight into the products obtained and their chemical structures. Regio- and stereochemical characterization and differentiation of each compound have been carried out in solution, in the crystal, and in the gas phase. A comparison between the data obtained in solution and the crystal structures suggests closely related features in the two states. Mass spectrometry proved very effective for characterization of and differentiation between C- and O-isomers, as well as positional isomers. Unimolecular reactions occurring in the gas phase are specific to the chemical structures, and ion abundances can be related to their stabilities. This study has allowed the evaluation of the influences of the different linkages between the two moieties and the positions of the substituents on the chemical properties of the compounds. The C-isomers show antioxidant capability, as peroxyl radical scavengers, and inhibit lipid peroxidation. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)

Synthesis of platelet-activating factor (PAF) in transformed cell lines of a different origin

Interest in the possible involvement of the platelet-activating factor (PAF) in tumor growth and invasiveness has been stimulated by the recognition that PAF influences various biological responses relevant to metastatic diffusion, such as angiogenesis, adhesiveness to endothelia and cellular motility. In the present study, we investigated the extent to which PAF is synthesized by a series of human and murine transformed cell lines of a different histotype. Synthesis of PAF was studied by combining the 14C-acetate incorporation into PAF with the quantitative analysis of PAF performed by a procedure based on gas chromatography-mass spectrometry with a negative ion chemical ionization. In the presence of the Ca2+ ionophore A23187, cultures of human melanoma (Hs294T), fibrosarcoma (HT1080) and colon carcinoma (LS180) cell lines synthesized conspicuous amounts of PAF, comparable to those produced by resident peritoneal macrophages. Substantial quantities of PAF were also synthesized by the murine melanoma (F10-M3 cells). PAF synthesis was rather limited in RSV-transformed Balb/c3T3 (B77-3T3) cells and in one of their high metastatic variants (B77-AA6 cells), although it was more abundant in the latter. We also investigated whether certain cytokines, such as TNFalpha and IFNgamma might induce PAF synthesis in our systems of cell lines which we found to express mRNAs encoding receptors for these cytokines. We observed that PAF synthesis was enhanced in human melanoma and colon carcinoma cell lines and in the murine B77-AA6 cells to levels comparable to those obtained with the Ca2+ ionophore. Synthesis of PAF was not inducible by TNFalpha in murine F10-M3 melanoma cells. IFNgamma also stimulated PAF synthesis in human and murine melanoma lines, and in human LS180 colon carcinoma line, but not in the B77-AA6 cells. PAF synthesis was also inducible by exogenous PAF in the human and murine melanoma lines, and in the human LS180 colon carcinoma line, all of which expressed cell surface PAF receptors. PAF synthesis was not inducible by exogenous PAF in the B77-AA6 cells, which do not express PAF receptors.

Structural characterization and differentiation of modified isomeric tryptophans

Different mass spectrometry (MS) techniques have been applied to the study of modified tryptophan isomers obtained by photochemical reactions. The gas phase behavior of the molecular ions and the most abundant fragment ions produced under electron ionization has been selectively studied by MS/MS experiments. Both the fragmentation reactions occurring in the ion source, as well as those produced under collision-induced dissociation conditions have allowed to characterize and differentiate each isomer from the others. Investigation of a bisubstituted derivative has been useful in the rationalization of the gas phase behavior of this series of modified tryptophans. This study has allowed the evaluation of the role played by the substituents and their positions at the indolic ring on the gas phase decompositions that are distinctive and selective for each isomer. The occurrence of regiospecific reactions suggests that isomerization phenomena do not occur either in the molecular ions or in the main fragment ions in the gas phase.

Molecular structure, characterization and stereochemical properties of new biologically interesting 3-(5-imidazo[2,1-b]thiazolylmethylene)-2-indolinones

Abstract New biologically interesting 3-(5-imidazo[2,1-b]thiazolylmethylene)-2-indolinones have been investigated by using X-ray crystallography, mass spectrometry, tandem mass spectrometry and theoretical calculations. The study has allowed to obtain information on the structural characterization of the stereoisomers and their conformational properties.

Application of self-ionization for enhancing stereochemical and positional effects from arylglycosides under electron ionization conditions in an ion trap mass spectrometer

Ion trap mass spectrometry has been used to structurally characterize and differentiate positional and stereo isomers of arylglycosides having potential antioxidant properties. The use of the self-ionization (SI) technique has allowed to evidence a strong reactivity of fragment ions produced from dissociations of the molecular ion towards the molecules introduced into the trap. Specific structural effects due to positional isomers and anomers have been also envisaged through the occurrence of bimolecular processes inside the ion trap analyzer. Under self-ionization conditions, even-electron ions are produced. The charge is retained on the sugar moiety, in agreement with its proton affinity higher than that of the substituted phenol moiety. Most of the fragmentation pathways involve elimination of acetic acid that protects the hydroxylic groups of the glycoside. SI also produces adduct ions, likely as covalent species, having higher m/z values than the molecular ion. The reaction site is mainly the double bond present in the pyranosidic ring. Even if some fragment ions have lost the initial stereochemistry, their formation can be related to the structure of the parent neutrals introduced into the cell. Collision-induced dissociation (CID) experiments, carried out on ions formed by ion-molecule reactions, have allowed to obtain further information on gas phase ion structures. The study of mass-selected ion-molecule reactions and their kinetics have evidenced a spectacularly different reactivity of the ion at m/z 111 towards the two anomers 2α and 2β, with the latter showing a much more pronounced reactivity. The approach developed in this work revealed to be an useful tool in structural characterization, as well as in stereo and regiochemical differentiation of arylglycosides.