Fatty Acids Profile of Wild and Cultivar Tunisian Peanut Oilseeds (A. hypogaea L.) at Different Developmental Stages.

Eleven fatty acids were identified during maturity in the wild (AraA) and varieties peanut kernels (AraC and AraT). These fatty acids included C16:0 (palmitic acid), C18:0 (stearic acid), C18:1 (oleic acid), C18:2 (linoleic acid), C19:0 (nonadecanoic acid), C20:1 (gadoleic acid), C20:0 (arachidic acid), C22:1 (erucic acid), C22:0 (behenic acid), C23:0 (tricosanoic acid) and C24:0 (linoceric acid). Two fatty acids C19:0 and C23:0 were not previously detected from peanut kernels. Furthermore, eight major fatty acids (C16:0, C18:0, C18:1, C18:2, C20:0, C20:1, C22:0 and C24:0) were quantified during maturity. Wild AraA was distinguished by its highest level of oleic (38.72%) and stearic (2.63%) acids contents and the lowest one of linoleic acid (19.40%) compared to the varieties. As for the O/L ratio, wild AraA presents a significantly higher (p < 0.05) (O/L = 2) than that of the AraC and AraT varieties with (O/L = 1.7 and 1.04) respectively. Correlation coefficients (r) between the eight major fatty acids revealed an inverse association between oleic and linoleic acids (r = -0.99, p < 0.001), while linoleic acid was positively correlated to palmitic acid (r = 0.97). These results aim to provide a detailed basis for quality improvement in the cultivated peanut with wild resources.

Theoretical and experimental studies of cationized uracil complexes in the gas phase.

Cationized uracil clusters were generated in the gas phase by electrospray ionization (ESI). Mass spectrometry experiments showed that with particular experimental conditions, decameric uracil clusters are magic number clusters. MS/MS experiments demonstrated that the structure of these decameric uracil clusters depends substantially on the size and the charge of the cation. On the basis of the ab initio and density functional theory (DFT) quantum chemistry calculations, structures for these decameric clusters were proposed. These structures are in agreement with the experimental mass spectra of modified nucleobases. Theoretical calculations showed that complexes experimentally observed using ESI-MS techniques, are not naturally the most stable in the gas phase.

Theoretical and experimental study of tropylium formation from substituted benzylpyridinium species.

Fragmentation pathways of unsubstituted and substituted benzylpyridinium compounds were investigated using mass-analysed kinetic energy (MIKE) technique in combination with high level of quantum chemical calculations in the gas phase. Fast atom bombardment (FAB) source was used for ionisation of the studied compounds. The formation of both benzylium and tropylium species were investigated. Hybrid Hartree-Fock/Density Functional Theory calculations have been performed to assess the geometries and the energies of the transition states and intermediates. For each cases, different reaction pathways were investigated, and particularly in the case of the formation of tropylium species, the formation of the seven-membered ring before or after the loss of pyridine were studied. The effect of para-methyl and para-methoxy substituents on the activation energy of the rearrangement process to form thermodynamically stable tropylium compounds has been studied. Theoretical calculations showed competition between direct bond cleavage and rearrangement reactions to form benzylium and tropylium compounds, respectively. Experimental results also suggested that the rearrangement process takes place to yield stable tropylium under "soft ionisation techniques", such as FAB.

Anionic copper complex fragmentations from enkephalins under low-energy collision-induced dissociation in an ion trap mass spectrometer.

Peptide metallation with Cu2+ was explored in the negative ESI mode using an ion trap mass spectrometer. Under these conditions, the [(M-3H) + CuII]- species formed were investigated under low-energy collision-induced dissociation conditions. MS2 experiments indicate a very different behavior of CuII metallated complexes compared with [M-H]- species. CuII induces an easy loss of CO2 and specific side-chain cleavages (by radical losses) at the C-terminal residue, as observed previously by prompt 'in source' dissociation experiments. The loss of CO2 yields an unstable carbylide that leads to further dissociations involving the migration of a proton or a hydrogen radical (through the reduction of CuII). Multistage MS3 experiments were carried out to rationalize this behavior. Fragmentation pathways are proposed in order to explain the product ions observed. The side-chain radical loss at the C-terminus was demonstrated to be a consecutive process. Finally, evidence is provided that the specific side-chain cleavages can be used for the differentiation of Leu/Ile and Gln/Lys residues when they are located at the C-terminus. The existence of a zwitterionic form in the case of the anionic YGGFK-CuII complex is proposed.

Changes in phospholipid composition, protein content and chemical properties of flaxseed oil during development.

The aim of the present research is to investigate the effect of harvest date on the composition of flaxseed. Samples were collected at regular intervals from 7 to 56 days after flowering (DAF) and analyzed for phospholipid composition, storage protein content and chemical properties. Phospholipid (PL) percentage of the total lipid decreased from 32.72% on the 7th DAF to 2.55% on the 56th DAF. The most phospholipids present in flaxseed were phosphatidylinositol (PI), phosphatidylethanolamine (PE) and lysophosphatidylcholine (LPC) which were highly unsaturated and rich in linolenic and linoleic acids, comprising together 60% of the total fatty acids. Chemical investigation of flaxseed oil showed overall a decrease in UV absorbance (K(232) and K(270)), acid value, free fatty acid content and an increase in peroxide value and storage protein content with development. At full maturity, flaxseed contained 29% proteins on a dry weight basis (DW %).

Electronic effects of 11ß substituted 17ß-estradiol derivatives and instrumental effects on the relative gas phase acidity.

Numerous studies have highlighted the role of the proton donor characteristics of the phenol group of 17ß-estradiol (E(2)) in its association with the estrogen receptor alpha (ERα). Since the substitutions at position C((11)) have been reported to modulate this association, we hypothesized that such substitutions may modify the phenol acidity. Hence, phenol gas-phase acidity of nine C((11))-substituted E(2)-derivatives were evaluated using the extended Cooks' kinetic method, which is a method widely used to determine thermochemical properties by mass spectrometry. To enhance accuracy in data collection we recorded data from several instruments, including quadrupole ion trap, triple quadrupole, and hybrid QqTOF. Indeed, we report for the first time the use of the QqTOF instrument to provide a novel means to improve data accuracy by giving access to an intermediate effective temperature range. All experimental gas-phase acidity values were supported by theoretical calculations. Our results confirmed the ability of distant substituents at C((11)) to modulate the phenol acidity through electrostatic interactions, electron withdrawing inductive effects, and mesomeric effects. However, no relationship was found between the phenol gas-phase acidity of investigated steroids and their binding affinity for ERα assessed in solution. Thus, our results highlight that the intrinsic properties of the hormone do not influence sufficiently the stabilization of the hormone/ERα complex. It is more likely that such stabilization would be more related to factors depending on the environment within the binding pocket such as hydrophobic, steric as well as direct intermolecular electrostatic effects between ERα residues and the substituted steroidal estrogens.

Non-thermal internal energy distribution of ions observed in an electrospray source interfaced with a sector mass spectrometer.

The internal energy distribution P(E(int)) of ions emitted in an electrospray (ESI) source interfaced with a sector mass spectrometer is evaluated by using the experimental survival yield (SY) method including the kinetic shift. This method is based on the relationship between the degree of fragmentation of an ion and its amount of internal energy and uses benzylpyridinium cations due to their simple fragmentation scheme. Quantum chemical calculations are performed, namely at G3(MP2)//B3LYP and QCISD/MP2 levels of theory. The results show that the internal energy distribution of the ions emitted in the ESI source interfaced with a sector analyzer is very narrow. The MassKinetics software is used to confirm these observations. The P(E(int)) is the parameter that allows to fit the experimental SY of each substituted benzylpyridinium cation with theoretical mass spectra generated by the MassKinetics software. The resulting internal energy distributions are similar to the ones obtained with the experimental SY method. This indicates that in the present experimental conditions, P(E(int)) cannot be compared with a 'thermal-like' Boltzmann distribution. In addition, it appears that with the sector analyzer, increasing the collision energy in the first pumping stage of the ESI source does not correspond to a warm-up of the produced ions.

Characteristics and pathways of bioactive 4-desmethylsterols, triterpene alcohols and 4α-monomethylsterols, from developing Tunisian cultivars and wild peanut (Arachis hypogaea L.).

Seven 4-desmethylsterols, five triterpene alcohols and three 4α-monomethylsterols were identified by GC-MS during the development of wild peanut, which is Arbi (AraA), and cultivars peanut, which are Trabelsia (AraT) and Chounfakhi (AraC). Our results showed that the maximum level of 4-desmethylsterols (881.07 mg/100 g of oil) was reached at 12 days after flowering (DAF) date of peanut plant in AraA, as well as the highest level of triterpene alcohols (31.51 mg/100 g of oil) was reached at 23 DAF in AraA, whilst, the highest level of 4α-monomethylsterols (15.11 mg/100 g of oil) was reached at 41 DAF in AraC. Herein, the level of triterpene alcohols and 4α-monomethylsterols was overwhelmed by the amount of 4-desmethylsterols at each stage of peanut maturity. Differences were observed in each sterol contents among the studied cultivars and wild one especially in immature stage.

Decameric uracil complexes around Li+.

Electrospray ionization (ESI) in combination with mass spectrometry (MS) experiments were carried out to study decameric uracil complexes cationized with Li(+) ion. A previous study has shown that, under specific experimental conditions, a particularly intense peak of the decamer U(10)Li(+) is formed, which was referred to as an indication for so-called 'magic number' cluster. In order to gain more insight on the structure of this decameric complex, here, we report experimental studies concerning the kinetics of the fragmentation. In accordance with the new experimental data, structural models were constructed and fully optimized using ab initio and density functional theory quantum chemistry calculations. The theoretical study allowed us to propose a stable gas-phase structure which is compatible with all experimental findings.

Energy-dependent dissociation of benzylpyridinium ions in an ion-trap mass spectrometer.

Benzylpyridinium ions, generated via electrospray ionization of dilute solutions of their salts in acetonitrile/water, are probed by collisional activation in an ion-trap mass spectrometer. From the breakdown diagrams obtained, phenomenological appearance energies of the fragment ions are derived. Comparison of the appearance energies with calculated reaction endothermicities shows a reasonably good correlation for this particular class of compounds. In addition, the data indirectly indicate that at threshold the dissociation of almost all of the benzylpyridinium ions under study leads to the corresponding benzylium ions, rather than the tropylium isomers. Substituent effects on the fragmentation for a series of benzylpyridinium ions demonstrate that neither mass effects nor differences in density of states seriously affect the energetics derived from the ion-trap experiments.

Gas chromatography-mass spectrometry screening for phytochemical 4-desmethylsterols accumulated during development of Tunisian peanut kernels (Arachis hypogaea L.).

4-Desmethylsterols, the main component of the phytosterol fraction, have been analyzed during the development of Tunisian peanut kernels ( Arachis hypogaea L.), Trabelsia (AraT) and Chounfakhi (AraC), which are monocultivar species, and Arbi (AraA), which is a wild species, by gas chromatography-mass spectrometry. Immature wild peanut (AraA) showed the highest contents of beta-sitosterol (554.8 mg/100 g of oil), campesterol (228.6 mg/100 g of oil), and Delta(5)-avenasterol (39.0 mg/100 g of oil) followed by peanut cultivar AraC with beta-sitosterol, campesterol, and Delta(5)-avenasterol averages of 267.7, 92.1, and 28.6 mg/100 g of oil, respectively, and similarly for AraT 309.1, 108.4, and 27.4 mg/100 g of oil, respectively, were found. These results suggest that, in immature stages, phytosterol contents can be important regulator factors for the functional quality of peanut oil for the agro-industry chain from plant to nutraceuticals.

Comparison of the concentrations of long-chain alcohols (policosanol) in three Tunisian peanut varieties (Arachis hypogaea L.).

Policosanol (PC) is a mixture of high molecular weight aliphatic primary alcohols. Literature about the contents and compositions of PC derived from peanut varieties is scarce. Total PC composition and content in whole peanut grain samples from three varieties of peanut (two cultivars, AraC and AraT, and a wild one, AraA) were identified using a gas chromatograph system coupled with a mass spectrophotometer. The results show that, qualitatively, 21 components of peanut aliphatic alcohols were identified (C14-C30). Besides (C18=), the results exhibited a previously unreported mixture of PC compositions in the peanuts: the unsaturated PC (UPC), which are (C20=), (C21=), (C22=), and (C24=). The main components of total PC in Tunisian peanut kernels are docosanol (C22), (Z)-octadec-9-en-1-ol (C18=), hexadecanol (C16), and octadecanol (C18). Quantitatively, the total PC content of the whole peanut samples varied from 11.18 to 54.19 mg/100 g of oil and was higher than those of beeswax and whole sugar cane, which are sources of dietary supplements containing policosanol.

Formation of complexes between uracil and calcium ions: an ESI/MS/MS study in combination with theoretical calculations.

Cationized uracil clusters around calcium metal ions were generated in the gas phase by electrospray ionization (ESI). A previous study showed that with particular experimental conditions, hexamer, octamer, decamer, dodecamer and tetradecamer uracil clusters are present in high quantities. New experiments were carried out to understand the reasons for the particular stability of these complexes. MS/MS experiments suggested that these uracil clusters belong to the same family. Based on ab initio and DFT quantum chemistry calculations, structures in agreement with experimental results are proposed for these clusters.

Characterization and quantification of the aliphatic hydrocarbon fraction during linseed development ( Linum usitatissimum L.).

Changes in hydrocarbon composition were investigated during maturation of three varieties of linseed (H52, O116, and P129) cultivated in Tunisia. The hydrocarbon fraction of the three linseed oil samples was found to contain mainly n-alkanes and squalene. The greatest decrease of these components occurred between 7 and 21 days after flowering (DAF); thereafter, the changes were slight. At 7 DAF, P129 had a significantly higher level of squalene (27.24 mg/100 g of oil) than H52 (3.36 mg/100 g of oil), but from this date until 21 DAF squalene decreased much more actively in P129, resulting in similar levels in H52 (0.57 mg/100 g of oil) and P129 (0.52 mg/100 g of oil) at full maturity. In three varieties of linseed, 13 n-alkanes were detected ranging from C(22) to C(34) carbon atoms. The n-alkane composition of linseed oil was influenced by the ripening stage of seeds. At 7 DAF, C29 was the most predominant hydrocarbon (19.84 mg/100 g of total oil), followed by C(27) (11.82 mg/100 g) and C(25) (11.28 mg/100 g). C(29) exhibited the most significant decrease from 7 to 21 DAF, as a result C(27) being the most significant n-alkane component for the remainder of the period. At full maturity, the content of total n-alkanes in three varieties of linseed ranged from 4.0 to 4.26 mg/100 g of oil.

Phytosterols accumulation in the seeds of Linum usitatissimum L.

A comparative study was performed to determine the free sterols content and composition during the development of three varieties of linseed (H52, O116 and P129). Seed samples were collected at regular intervals from 7 to 60 days after flowering (DAF). Ten compounds were identified: cholesterol, campesterol, brassicasterol, stigmasterol, beta-sitosterol, Delta5-avenasterol, cycloartenol; 24-methylene cycloartanol, obtusifoliol, citrostadienol. The maximum level of 4-desmethylsterols (1,515 mg/100g oil) was reached at 7 DAF in P129 variety. H52 had the highest level of 4-4 dimethylsterols (355 mg/100g oil) at 28 DAF. The greatest amount of 4-monomethylsterols (35 mg/100g oil) was detected in H52 at 14 DAF. During linseed development, beta sitosterol (830 mg/100g oil) was the major 4-desmethylsterols, followed by campesterol (564 mg/100g oil) and stigmasterol (265 mg/100g oil). Some of these compounds followed nearly the same accumulation pattern during linseed maturation.

Investigations of the fragmentation pathways of benzylpyridinium ions under ESI/MS conditions.

Benzylpyridinium ions are often used as 'thermometer ions' in order to evaluate the internal energy distribution of the ions formed in sources of mass spectrometers. However, the detailed fragmentation pathways of these parent ions were not well established. In particular, fragmentation involving a rearrangement (RR) process may be influencing the simulated distribution curves. In a previous study, we suggested that such RR actually occurred under electrospray ionization/mass spectrometry (ESI/MS) and fast atom bombardment/mass spectrometry (FAB/MS) experiments. Here, we present a systematic study of different substituted benzylpyridinium ions. Theoretical calculations showed that RR fragmentation leading to substituted tropylium ions could occur under 'soft ionization' conditions, such as ESI or FAB. Experimental results obtained under gas-phase reactivity conditions showed that some substituted benzylpiridinium compounds actually undergo RR fragmentations under ESI/MS conditions. Mass-analyzed kinetic experiments were also carried out to gain information on the reaction pathways that actually occur, and these experimental results are in agreement with the reaction pathways theoretically proposed.

Stereochemical effects during [M-H]- dissociations of epimeric 11-OH-17beta-estradiols and distant electronic effects of substituents at C(11) position on gas phase acidity.

The affinity of estradiol derivatives for the estrogen receptor (ER) depends strongly on nature and stereochemistry of substituents in C(11) position of the 17beta-estradiol (I). In this work, the stereochemistry effects of the 11alpha-OH-17beta-estradiol (III(alpha)) and 11beta-OH-17beta-estradiol (III(beta)) were investigated using CID experiments and gas-phase acidity (DeltaH degrees (acid)) determination. The CID experiments showed that the steroids decompose via different pathways involving competitive dissociations with rate constants depending upon the alpha/beta C(11) stereochemistry. It was shown that the fragmentations of both deprotonated [III(alpha)-H]- and [III(beta)-H]- epimers were initiated by the deprotonation of the most acidic site, i.e. the phenolic hydroxyl at C(3). This view was confirmed by H/D exchange and double resonance experiments. Furthermore, the DeltaH degrees (acid) of both epimers (III(alpha) and III(beta)), 17beta-estradiol (I), and 17-desoxyestradiol (II) was determined using the extended Cooks' kinetic method. The resulting values allowed us to classify steroids as a function of their gas-phase acidity as follows: (III(beta)) >> (II) > (I) > (III(alpha)). Interestingly, the alpha/beta C(11) stereochemistry appeared to influence strongly the gas-phase acidity. This phenomenon could be explained through stereospecific proton interaction with pi-orbital cloud of A ring, which was confirmed by theoretical calculation.

Theoretical and gas-phase studies of specific cationized purine base quartet.

Guanine tetraplexes are biological non-covalent systems stabilized by alkali cations. Thus, self-clustering of guanine, xanthine and hypoxanthine with alkali cations (Na(+), K(+) and Li(+)) is investigated by electrospray ionization mass spectrometry (ESI-MS) in order to provide new insights into G-quartets, hydrogen-bonded complexes. ESI assays displayed magic numbers of tetramer adducts with Na(+), Li(+) and K(+), not only for guanine, but also for xanthine bases. The optimized structures of guanine and xanthine quartets have been determined by B3LYP hybrid density functional theory calculations. Complexes of metal ions with quartets are classified into different structure types. The optimized structures obtained for each quartet explain the gas-phase results. The gas-phase binding sequence between the monovalent cations and the xanthine quartet follows the order Li(+) > Na(+) > K(+), which is consistent with that obtained for the guanine quartet in the literature. The smallest stabilization energy of K(+) and its position versus the other alkali metal ions in guanine and xanthine quartets is consistent with the fact that the potassium cation can be located between two guanine or xanthine quartets, for providing a [gua(or (xan))(8)+K](+) octamer adduct. Even if an abundant octamer adduct with K(+) for xanthine was detected by ESI-MS, it was not the case for guanine.