Publications (30)63.43 Total impact
- [Show abstract] [Hide abstract] ABSTRACT: The modification of arginine residues by diketonic α-dicarbonyls, in structural proteins and enzymes studies, is a process known for decades. The chemistry of these reaction processes is, however, not fully understood. Moreover, modification of arginine residues by reaction with α-dicarbonyls in glycation has also not been completely elucidated. Aspects related to the early stages of the condensation of arginine with one dicarbonyl molecule, leading to the formation of dihydroxyimidazolidines and their dehydrated forms, in particular, are here approached in more detail. Taking into consideration the usually rapid kinetics involved in the formation of the early reaction product species, we decided to use fast, sensitive and selective analytical techniques, such as electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) to monitor the reactions of a blocked arginine (acetyl-arginine) with several selected diketonic α-dicarbonyls, to identify and characterize the mentioned transient species and to probe the reaction mechanism involved. Compounds grouped into two different classes according to their structural similarity were identified, namely acetyl-dihydroxyimidazolidines and acetyl-bis(dihydroxyimidazolidines), together with their dehydrated species. The former compounds are known to exist in solution. The reactivity of acetyl-bis(dihydroxyimidazolidines) seems to be different from that of acetyl-dihydroxyimidazolidines. To note that dehydration appears to be reinforced in acetyl-bis(dihydroxyimidazolidines) chemistry with respect to acetyl-dihydroxyimidazolidine chemistry, while both structurally related compounds involve mostly dihemiaminals reactivity. Two different ion structures are proposed for single dehydrated acetyl-bis(dihydroxyimidazolidines), concerning the two more symmetrical and two more asymmetrical dicarbonyls reacted. In acetyl-bis(dihydroxyimidazolidines) formation, we concluded that the importance of single dehydration relies on the rapid minimization of sterics and energetics of the reaction moieties formed. These reactions occur also in a selective way, regarding the two compound structures proposed for single dehydrated acetyl-bis(dihydroxyimidazolidines). Further considerations are also established for the formation of single dehydrated acetyl-bis(dihydroxyimidazolidines). An explanation for the reversible nature of the reaction of arginine with diketonic dicarbonyls is also provided. This study reinforces the potential of the fast, sensitive and selective electrospray ionization mass spectrometry techniques for the investigation of transient species and their mechanistics, that might otherwise not be feasible by means of the most commonly used spectroscopic techniques.
- [Show abstract] [Hide abstract] ABSTRACT: The inhibition of α-glucosidase and glucose-6-phosphatase, two enzymes involved in the carbohydrate metabolism, is an important target to control glycaemia on individuals with type 2 diabetes. In this work we report for the first time the inhibition of both enzymes by the antihyperglycemic n-butanol extract from Genista tenera (Fabaceae). This extract decreased α-glucosidase and glucose-6-phosphatase activities to 0.97 and 80.25 %, respectively, being more effective than acarbose, and phlorizin, the positive controls, which reduced enzymes activities only to 17.39 and 96.06 %. Once inflammation and oxidative stress are related to diabetic impairments, the anti-inflammatory activity of the extract was also evaluated, through its inhibitory activity over COX-1 enzyme (47.5 % inhibition). Moreover, after induction of oxidative stress by UV radiation, the viability of irradiated rat liver hepatoma cells exposed to the extract was significantly higher (67.82 %) than that promoted by ascorbic acid, the positive control (45.05 %). In addition, the stability of the extract under gastrointestinal conditions was evaluated by HPLC–DAD-ESI–MS/MS. Flavonoid diglycosides were identified as the main constituents of the extract, and no alterations in the chemical composition nor in the antioxidant activity were observed after in vitro digestion with artificial gastric and pancreatic juices. Graphical Abstract
- [Show abstract] [Hide abstract] ABSTRACT: Aminoguanidine possesses extensive pharmacological properties. This drug is recognized as a powerful α-dicarbonyl scavenger. In order to better elucidate the reactivity of aminoguanidine with α-dicarbonyls, aminoguanidine was reacted with several aldehydic and diketonic α-dicarbonyls. Electrospray ionization mass spectrometry is a suitable technique to study chemical and biochemical processes, and was selected for the purpose. In aminoguanidine reactions, triazines were detected and, other compounds that have never been reported before were identified. Triazine precursor forms were detected, namely tetrahydrotriazines and singly dehydrated tetrahydrotriazines. Moreover, species with bicyclic ring structures, and dehydrated forms, were also identified in aminoguanidine reactions. These species appear to result from tetrahydrotriazines and triazines reactions with one dicarbonyl molecule. Experiments revealed that these bicyclic species, in particular the ones resulting from triazines reactivity, could exist in solution, since they were both identified in the reactions of aminoguanidine and of a selected triazine with the dicarbonyls studied. The results obtained, regarding aminoguanidine/triazines reactivities, appear to support the capability of triazines to condensate and form polycyclic ring structures, and also to support literature mechanistic data for dihydroimidazotriazines formation via dihydroxyimidazolidine-triazines. The data obtained in this study may prove to be valuable to complement solution information, concerning the reactivity of amines with α-dicarbonyls, in particular.
- [Show abstract] [Hide abstract] ABSTRACT: Five isoflavones, daidzein, genistein, formononetin, prunetin and biochanin A, known for their biological properties, are investigated by electrospray ionization mass spectrometry in the positive ion mode. The most probable protonation sites are determined taking into account semi-empirical calculations using the PM6 Hamiltonian. Fragmentation mechanisms are proposed based on accurate mass measurements, MS(3) experiments and supported by the semi-empirical calculations. Some of the fragmentation pathways were found to be dependent on the substitution pattern of the B-ring and the ions afforded by these fragmentations can be considered as diagnostic. It was possible to distinguish between prunetin and biochanin A, two isobaric isoflavone aglycones included in this study. Furthermore, a comparison of the fragmentation patterns of genistein and biochanin A, two isoflavones, with those of their flavone counterparts, apigenin and acacetin, enabled us to identify some key ions mainly due to structural features, allowing distinction to be made between these two classes of compounds.
- [Show abstract] [Hide abstract] ABSTRACT: ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
- [Show abstract] [Hide abstract] ABSTRACT: The antihyperglycaemic effect of eight standard flavonoids, previously identified in the ethanol extract of the claimed antidiabetic plant Genista tenera, was evaluated on streptozotocin (STZ)-induced diabetic Wistar rats. The aglycones apigenin, chrysoeriol and genistein, the monoglucosides apigenin 7-O-glucoside, luteolin 7-O-glucoside and genistein 7-O-glucoside and the diglycosides rutin and luteolin 7,3'-di-O-glucoside were administered i.p. for 7 days (4 mg/kg b.w./day). The protective effect of these compounds over liver and kidneys of STZ-diabetic models was also evaluated by the determination of seric AST, ALT and urea levels. After 7 days of treatment, apigenin, chrysoeriol and genistein significantly lowered the blood glucose levels of diabetic animals; this effect was more pronounced (P < 0.01) in the oral glucose tolerance test. Glucose tolerance was also significantly improved in the rutin (P < 0.01) and in the genistein 7-O-glucoside (P < 0.05) treated groups. In addition, almost all the tested compounds effectively protected the liver and kidneys against STZ-induced damage in rats.
- [Show abstract] [Hide abstract] ABSTRACT: Our previous experiments on ESI-MS analysis of reaction mixture solutions containing HEPES (4-(-2- hydroxyethyl)-1-piperazineethanesulfonic acid), a commonly used buffer, indicated that HEPES species did not significantly suppress analyte species, even in reaction mixture solutions with significant amounts of HEPES. With the purpose of investigating the behaviour of HEPES under ESI-MS conditions, HEPES aqueous solutions and HEPES aqueous solutions containing analyte with high and low polarity and with different acid/base chemistry, were therefore investigated. For electrosprayed aqueous solutions of HEPES with concentrations above 10(-5) M, an enhanced formation of HEPES multimer ions, regarding HEPES monomer ions formation, was observed. This enhanced formation of HEPES multimer ions is much higher than the one observed for other polar compounds, such as acetyl-arginine, acetyl-lysine and histidine. Information from solution behaviour such as, HEPES concentration, solution pH, and instrumental factors, namely the capillary temperature, was related with information from mass spectra. The results obtained led us to conclude that the formation of HEPES ions is related with the initial solution composition. The influence of analyte species on HEPES species formation, for electrosprayed HEPES solutions with analyte, was also investigated. The variations observed for HEPES monomer and multimer ions abundances, which were found to be consistent with those observed for analyte monomer ions abundances, were related with type of analyte, i.e. to their acid/base nature. Strikingly, the variations observed between HEPES monomer and multimer ions abundances, enable to discriminate among the different influence of analyte species on HEPES species formation. The results obtained also enabled to provide an explanation for the observation that HEPES species do not suppress significantly analyte species ion signals, when high concentrated HEPES solutions with analyte are electrosprayed. According to our results, the association behaviour between HEPES species seems to be preserved in the gas phase during electrospray ionization. This observation may provide some information that may be useful regarding the behaviours involved in the gas phase ion formation process from charged droplets during electrospray ionization or, at least, to differentiate among behaviours.
- [Show abstract] [Hide abstract] ABSTRACT: Genista tenera is a plant endemic to the island of Madeira and is used in folk medicine to control diabetes. In the present work we evaluate the antihyperglycaemic activity of its n-butanol extract and determine its chromatographic profile. In addition, this extract, the ethyl acetate and diethyl ether plant extracts were studied in order to assess the plant antioxidant and acetylcholinesterase inhibitory activities, as well as its cyto- and genotoxicities. HPLC-DAD-ESI-MS was used to analyze the flavonoid profile of the n-butanol extract. The antihyperglycaemic activity of this extract was performed over streptozotocin induced diabetic Wistar rats (200 mg/kg, bw/day), for 15 days. Antioxidant activity (DPPH assay) and acetylcholinesterase inhibitory effect (Ellman method) were also performed. Acute cytotoxicity and genotoxicity were assessed by proliferative index quantification and the short-term chromosomal aberration technique, after exposure of lymphocytes to the extracts. The n-butanol extract, where 21 monoglycosyl and 12 diglycosyl flavonoids were detected, significantly lowered blood glucose levels, bringing them to normal values after 15 days of treatment. The best radical scavenging activity was observed for the ethyl acetate extract (48.7% at 139.1 microg/mL), which was also the most effective one at the minimal concentration tested. The highest acetylcholinesterase inhibitory activity (77.0% at 70.0 microg/mL) was also obtained with the ethyl acetate extract. In vitro toxicity studies showed no evidence for acute cytotoxicity or genotoxicity. This is the first report on antidiabetic activity of genus Genista.
- [Show abstract] [Hide abstract] ABSTRACT: Flavonoids are important phytochemicals which have been intensively studied in the last decades in view of their antioxidant activity, which is of particular importance in the case of flavones and flavonols, that differ in a single 3-OH group. Mass spectrometry has been used to elucidate the structures of many types of flavonoids and their metabolites. The work we present here is focused on the electrospray ionization tandem mass spectrometry (ESI-MS/MS) analysis of flavone and flavonols aglycones. Their fragmentation mechanisms in the positive ion mode are described and compared with previously reported mechanisms. We analyzed flavonoid derivatives produced by reaction of the flavonoids with chemically synthesized hypohalous acids (HOCl, HOBr and HOI) and peroxynitrite, reactive species involved in the inflammatory response. All the proposed pathways have been analyzed using computational chemistry methods in order to seek for possible variations and establish the most plausible ones. We observed that the losses of one and two CO molecules can be useful in terms of antioxidant activity prediction. Losses of one and two C(2)H(2)O groups are also informative in terms of structure and activity predictions. The retro-Diels-Alder fragmentations, and subsequent neutral losses, were reviewed and, according to our calculations, the most plausible structures for the product ions were established. These fingerprints will be of great value for differentiating flavonoids from other compounds in complex biological mixtures and for a thorough structural identification of flavonoid aglycones and their in vivo metabolites.
- [Show abstract] [Hide abstract] ABSTRACT: In Saccharomyces cerevisiae, the rate of hydrogen peroxide (H(2)O(2)) diffusion through the plasma membrane decreases during adaptation to H(2)O(2) by a still unknown mechanism. Here, adaptation to H(2)O(2) was observed to modulate rapidly the expression of genes coding for enzymes involved in ergosterol and lipid metabolism. Adaptation to H(2)O(2) also alters plasma membrane lipid composition. The main changes were the following: (a) there was a decrease in oleic acid (30%) and in the ratio between unsaturated and saturated long-chain fatty acids; (b) the phosphatidylcholine:phosphatidylethanolamine ratio increased threefold; (c) sterol levels were unaltered but there was an increased heterogeneity of sterol-rich microdomains and increased ordered domains; (d) the levels of the sterol precursor squalene increased twofold, in agreement with ERG1 gene down-regulation; and (e) C26:0 became the major very long chain fatty acid owing to an 80% decrease in 2-hydroxy-C26:0 levels and a 50% decrease in C20:0 levels, probably related to the down-regulation of fatty acid elongation (FAS1, FEN1, SUR4) and ceramide synthase (LIP1, LAC1) genes. Therefore, H(2)O(2) leads to a reorganization of the plasma membrane microdomains, which may explain the lower permeability to H(2)O(2), and emerges as an important regulator of lipid metabolism and plasma membrane lipid composition.
- [Show abstract] [Hide abstract] ABSTRACT: The effect of an aqueous extract of Pterospartum tridentatum on the blood glucose levels of normal Wistar rats was investigated in a situation of oral glucose challenge. The extract at 300 mg/kg showed an antihyperglycaemic effect in the first 30 min after glucose challenge but then the blood glucose levels rose above those of the control group, indicating the presence of compounds with different effects on glucose tolerance. Nine compounds of isoflavone and flavonol skeletons were identified in the extract by HPLC-ESI-MSn, four of them being identified for the first time in this species. The isoflavone sissotrin and the flavonol derivative, isoquercitrin, were selected for the oral glucose tolerance test. Isoquercitrin (100 mg/kg) showed time-dependent antihyperglycaemic activity by delaying the post-oral glucose load glycaemic peak at 30 min, as did the sodium-dependent glucose transporter inhibitor phloridzin (100 mg/kg). In contrast, sissotrin (100 mg/kg) showed an opposite effect, impairing glucose tolerance. In conclusion, these preliminary results indicate that the effect of the extract on blood glucose may be either antihyperglycaemic or hyperglycaemic. Additionally, as far as is known, these are the first in vivo results on the acute antihyperglycaemic potential of isoquercitrin. Copyright © 2008 John Wiley & Sons, Ltd.
- [Show abstract] [Hide abstract] ABSTRACT: Glycation of proteins by glucose and formation of end-stage adducts (AGEs, advanced glycation end products) has been implicated in pathological mechanisms associated with diabetic complications, macrovascular disease, chronic and renal insufficiency, Alzheimer's disease, and aging. Of the carbonyl containing compounds involved in this process, α-dicarbonyls have particular importance, being established as direct intermediates in the formation of well-known AGEs. The guanidino group, present in arginine residues, suffers direct modifications by sugars and its derivatives, and is considered to be an important chemical basis, targeting the control and inhibition of glycation. Seven dicarbonyl compounds, aldehydic and diketonic, were reacted with guanidine, in an attempt to establish structure/activity relationships. Electrospray mass spectrometry, together with tandem mass spectrometry, was used to identify and characterize the reaction products. The reactivity of guanidine was found to vary with the dicarbonyls used. For glyoxal, a high amount of dihydroxyimidazolidine was formed, whereas for methylglyoxal, dihydroxyimidazolidine was slowly converted into hydroimidazolone. Interestingly, aqueous guanidine was found to prevent argpyrimidine formation. The formation of several amine-dicarbonyl moieties was observed for the larger alkyl-diketonic dicarbonyls reaction systems, in particular. Molecular structures, bearing a polar chain, of an imidazole ring, and a nonpolar one, of alkyl groups, located at both sides of the imidazole rings, were attributed to these moieties. Gas-phase experiments suggested that the larger alkyl groups have a preference for being located at one of the sides of the imidazole rings. Moreover, the referred amine-dicarbonyl moieties are formed via (dihydroxyimidazolidine − 2H2O) moieties. The latter (dihydroxyimidazolidine − 2H2O) moieties are formed in high amounts in the larger alkyl-diketonic dicarbonyl reactions. Since these moieties react with dicarbonyl molecules, and react even faster with already modified amine functions, we can foresee that these species may be useful for controlling and inhibiting glycation of larger biomolecules, such as proteins. Copyright
- [Show abstract] [Hide abstract] ABSTRACT: Non-enzymatic glycation (Maillard reaction) of long-lived proteins is a major contributor to the pathology of diabetes, and possibly aging and Alzheimer's disease. Among the amino residues in proteins arginine plays an important role, and its modification by sugar moieties generates the so-called advanced glycation end products (AGEs). Moreover, α-dicarbonyl compounds have been found as the main participants in those modifications. Four α-dicarbonyl compounds, aldehydic and ketonic, were reacted with the modified amino acid Nα -acetyl-L-arginine (AcArg), in an attempt to establish structure/activity relationships for the reactivity of α-dicarbonyls with the amine compound. Electrospray ionization mass spectrometry (ESI-MS), combined with tandem mass spectrometry (MS/MS), was used to identify and characterize reagents, intermediates and reaction products. The fragmentation patterns of precursor ions showed similarities in all reaction systems studied, in which fragmentation of the amino acid residue prevails, especially for the dehydrated and/or multiple dehydrated precursor ions. For the non-hydrated ion species, fragmentation of the arginyl guanidino group was mainly observed. Specific information regarding the nature of the ions formed, in which the dicarbonyl electrophile character played an important role, was obtained. As an example, singly and doubly hydrated acetyl-argpyrimidine ions were detected for the methylglyoxal reaction only. For symmetrical dicarbonyls, glyoxal and diacetyl, the importance of steric contributions with respect to the energetic ones is discussed. Furthermore, the dehydrated acetyl-tetrahydropyrimidine ions for methylglyoxal and phenylglyoxal reactions revealed fragment ion compositions including the protonated molecules of acetyl-argpyrimidine, -hydroimidazolone and -5-methylimidazolone. An explanation for the acetyl-argpyrimidine formation from the acetyl-hydroimidazolone formation reaction is proposed. Aspects such as the amount of acetyl-hydroimidazolone formed, the response of the hydration equilibria of the dicarbonyl forms to the new unhydrated dicarbonyls introduced by the reversal of the acetyl-hydroimidazolone formation reaction and the stability of the dicarbonyl intermediate involved in the acetyl-argpyrimidine formation are proposed, as being responsible to control the formation of acetyl-argpyrimidine. Copyright
- [Show abstract] [Hide abstract] ABSTRACT: The phenomenon known as non-enzymatic glycation is described as the reaction of reducing sugars with basic amino groups of proteins and nucleic acids, as well as with simple amines, without enzyme mediation. Non-enzymatic model glycation reactions that make use of low-molecular-weight compounds make an important contribution in the elucidation of glicated processes in vitro and in vivo. Four alpha-dicarbonyl compounds, aldehydic (glyoxal, methylglyoxal and phenylglyoxal) and ketonic (diacetyl), were reacted with the modified amino acid N(alpha)-acetyl-L-lysine (AcLys) in an attempt to establish structure/activity relationships for the reactivity of alpha-dicarbonyls with the amine compound. Electrospray ionization mass spectrometry (ESI-MS) combined with tandem mass spectrometry (MS/MS) and collision-induced dissociation (CID) was used to identify and characterize reagents, intermediates and reaction products. The formation of dicarbonyl-derived lysine dimers was observed exclusively. Especially, attention is drawn to alkyl- (asymmetrical dicarbonyl systems) and carboxyl- (glyoxal system) substituted imidazolium ions, at ring position 2. The main differences observed in the reactions studied were related to the reactivity with the diimine intermediate. This intermediate can react either with a non-hydrated dicarbonyl molecule at the aldehydic carbonyl, or with a mono-hydrated one at the ketonic carbonyl, particularly for asymmetrical dicarbonyls. For 2-carboxyl-substituted imidazolium ion (glyoxal reaction), besides the usual keto-enol rearrangement from the diol group, an alternative reaction pathway (proton abstraction) appears to contribute also for the imidazolium ring-closure process. Moreover, the formation of imidazolium ring structures can depend on several factors, namely, the presence (or absence) of electron donor substituents at the formed diol, the degree of stability of the new electrophile generated and/or the equilibrium concentration of the non- and mono-hydrated dicarbonyl forms in solution, the last being particularly important for asymmetrical dicarbonyls. The results reported reveal the complexity of reactivity as well as the diversity of imidazolium molecular structures.
- [Show abstract] [Hide abstract] ABSTRACT: The spectral data of a new Delta7,22 sterol, 24S-ethyl-5alpha-cholesta-7,22E-dien-3alpha-ol beta-galactopyranoside, isolated from the bulbs of Autonoë madeirensis, are reported.
- [Show abstract] [Hide abstract] ABSTRACT: The anti-hyperglycemic flavonoid extract obtained from Genista tenera was first studied by liquid chromatography (LC)-diode array detection (DAD) which showed the presence of two major compounds. One of them was identified as genistein-7-O-glucoside. Luteolin-7-O-glucoside was detected as a minor constituent, while luteolin-7,3'-di-O-glucoside and rutin were found in trace amounts. LC-DAD-ESI-MS and NMR were used to confirm the structure of these compounds and allowed the elucidation of the structure of the unknown major compound, which is the flavonoid 5,7,4'-trihydroxyisoflavone-8-C-glucoside.
- [Show abstract] [Hide abstract] ABSTRACT: The benzidine photodegradation process was studied using UV/Vis spectroscopy and electrospray ionization mass spectrometry (ESI-MS) combined with collision-induced dissociation (CID) and tandem mass spectrometry (MS/MS). Mass spectrometry was used to characterize benzidine and to identify and characterize possible degradation products and intermediates. The MS data showed that benzidine is quite persistent in aqueous medium. Moreover, the MS analysis enabled us to propose the following three degradation products/intermediates: 4'-nitro-4-biphenylamine, tetrahydroxybiphenyl and 4,4'-dinitrobiphenyl. For the benzidine molecular ion and protonated molecule and for the protonated molecules of the degradation products/intermediates detected, fragmentation patterns are proposed based on CID and MS/MS data. For the photodegradation process different catalysts were used, namely the commercial TiO2 Degussa P25, and the laboratory-prepared ZnO, TiO2 anatase and a titanium-zinc oxide with a perovskite type structure. Comparison of the different catalysts showed that degradation was favoured with the commercial TiO2. Nevertheless, the other catalysts appear to be promising and economic alternatives for potential future remediation studies.
- [Show abstract] [Hide abstract] ABSTRACT: We studied quercetin metabolism in rats to determine the nature and conjugation positions on the resulting metabolites and to evaluate their contribution to the antioxidant activity of plasma. HPLC analysis showed that quercetin is primarily metabolized to glucuronides and sulfoglucuronides and, to a minor extent, to sulfates. ESI-MS/MS studies confirmed these results and indicate that the most plausible positions for glucuronidation and sulfation are the hydroxyl groups located at positions 5 and 7, excluding the 3'-OH and 4'-OH groups. Plasma antioxidant status was significantly higher in animals to which quercetin was administrated, suggesting that quercetin metabolites can retain some antioxidant activity when the o-catechol group does not undergo conjugation reactions. It was also shown that plasma quercetin metabolites could compete in vivo with other molecules for peroxynitrite. These results enabled the establishment of quercetin metabolite structure-antioxidant activity relationships and, hence, to understand their contribution for the antioxidant potential of plasma.
University of Lisbon
Lisboa, Lisbon, Portugal
- Faculty of Science