Susana M. Cardoso

Phenolic characterization of Northeast Portuguese propolis: usual and unusual compounds.

In this study, an ethanolic extract from Portuguese propolis was prepared, fractionated by high-performance liquid chromatography, and the identification of the phenolic compounds was done by electrospray mass spectrometry in the negative mode. This technical approach allowed the identification of 37 phenolic compounds, which included not only the typical phenolic acids and flavonoids found in propolis from temperate zones but also several compounds in which its occurrence have never been referred to in the literature. Four of the novel phenolic compounds were methylated and/or esterified or hydroxylated derivatives of common poplar flavonoids, although six peculiar derivatives of pinocembrin/pinobanksin, containing a phenylpropanoic acid derivative moiety in their structure, were also identified. Furthermore, the Portuguese propolis sample was shown to contain a p-coumaric ester derivative dimer.

Calcium-mediated gelation of an olive pomace pectic extract

Pectic raw material was extracted from the alcohol insoluble residue of olive pomace. After purification, the olive pectic extract (OPE) contained 48% of galacturonic acid (GalA) and 31% of arabinose, in a total sugar content of 72%, and a degree of methylesterification (DM) of 43%. Phase diagrams were established to define the physical state of the OPE/calcium systems, at pH 3 and 7, as a function of GalA and calcium concentration. The rheological properties of the OPE/calcium systems were investigated by small-amplitude oscillatory tests. Kinetics of gel ageing and the viscoelastic properties of the cured gels were evaluated as a function of GalA and calcium concentration at pH 3 and 7. Compared with the observed behaviour of a citrus pectin, taken as representative of a commercial low-methoxyl pectin, the OPE showed higher critical GalA and calcium concentrations for gelation to occur, a larger region corresponding to homogeneous gel, and gels characterised by lower viscoelastic moduli, at similar GalA and calcium concentrations.

Temperature dependence of the formation and melting of pectin-Ca2+ networks: a rheological study

Abstract Small deformation dynamic rheometry was used to characterise the calcium-induced gelation of low-methoxyl pectins, at two different pH values. The gelation kinetics was interpreted with basis on the change of the storage modulus with time, taken as a measure of changes in cross-linking density within the pectin–calcium network. The temperature influence on the rate of gel formation and ageing was evaluated, as well as the temperature sensitivity of the cured gels. The formation and the softening/melting of pectin–calcium networks were also studied under non-isothermal conditions. The structural diversity of the two pectin samples had a great influence on the gelation kinetics and thermal behaviour of these pectin–calcium networks, due to differences in the steric arrangement or environment and/or availability of the chelating groups. An association mechanism is suggested to predominate under conditions of low availability of dissociated carboxylic groups, due to a low pH, higher degree of methylation, or steric constraints introduced by acetylation or neutral side chains, different from the classical ‘egg-box’ model, and where non-ionic hydrogen bonding and hydrophobic interactions may co-exist and cooperate with coordinative binding of calcium-ions.

Structural characterisation of the olive pomace pectic polysaccharide arabinan side chains

An arabinan (97% of Ara and 3% of hexuronic acid) was isolated from the alcohol-insoluble residue (AIR) of olive pomace by treatment with 0.02 M HNO(3), at 80 degrees C, followed by graded precipitation with ethanol. It was separated from acidic pectic polysaccharides by anion-exchange chromatography, and by size-exclusion chromatography its molecular weight was estimated as 8.4 kDa. By methylation analysis, the linkage composition was established as 5:4:3:1 for (1-->5)-Araf, T-Araf, (1-->3,5)-Araf and (1-->3)-Araf, respectively. 13C NMR spectroscopy confirmed this linkage composition, and allowed to assign the alpha anomeric configuration for the arabinofuranosyl residues, except for some terminally linked ones, that were seen to occur as T-beta-Araf. By 2D NMR spectroscopy (1H and 13C), it was possible to conclude that the T-beta-Araf was (1-->5)-linked to a (1-->5)-Araf residue. Also, in the arabinan (1-->5)-Araf backbone, the branched (1-->3,5)-Araf residues were always adjacent to linear (1-->5)-Araf residues. A tentative structure is proposed.

Phenolic Profiling of Portuguese Propolis by LC-MS Spectrometry: Uncommon Propolis Rich in Flavonoid Glycosides

INTRODUCTION Propolis is a chemically complex resinous substance collected by honeybees (Apis mellifera) from tree buds, comprising plant exudates, secreted substances from bee metabolism, pollen and waxes. Its chemical composition depends strongly on the plant sources available around the beehive, which have a direct impact in the quality and bioactivity of the propolis. Being as Portugal is a country of botanical diversity, the phenolic characterisation of propolis from the different regions is a priority. OBJECTIVE Extensive characterisation of the phenolic composition of Portuguese propolis from different continental regions and islands. METHOD Forty propolis ethanolic extracts were analysed extensively by liquid chromatography with diode-array detection coupled to electrospray ionisation tandem mass spectrometry (LC-DAD-ESI-MS(n) ). RESULTS Seventy-six polyphenols were detected in the samples and two groups of propolis were established: the common temperate propolis, which contained the typical poplar phenolic compounds such as flavonoids and their methylated/esterified forms, phenylpropanoid acids and their esters, and an uncommon propolis type with an unusual composition in quercetin and kaempferol glycosides - some of them never described in propolis. CONCLUSION The method allowed the establishment of the phenolic profile of Portuguese propolis from different geographical locations, and the possibility to use some phenolic compounds, such as kaempferol-dimethylether, as geographical markers. Data suggest that other botanical species in addition to poplar trees can be important sources of resins for Portuguese propolis.

Seaweeds as Preventive Agents for Cardiovascular Diseases: From Nutrients to Functional Foods

Being naturally enriched in key nutrients and in various health-promoting compounds, seaweeds represent promising candidates for the design of functional foods. Soluble dietary fibers, peptides, phlorotannins, lipids and minerals are macroalgae’s major compounds that can hold potential in high-value food products derived from macroalgae, including those directed to the cardiovascular-health promotion. This manuscript revises available reported data focusing the role of diet supplementation of macroalgae, or extracts enriched in bioactive compounds from macroalgae origin, in targeting modifiable markers of cardiovascular diseases (CVDs), like dyslipidemia, oxidative stress, vascular inflammation, hypertension, hypercoagulability and activation of the sympathetic and renin-angiotensin systems, among others. At last, the review also describes several products that have been formulated with the use of whole macroalgae or extracts, along with their claimed cardiovascular-associated benefits.

Oleuropein/ligstroside isomers and their derivatives in Portuguese olive mill wastewaters

Olive mill wastewaters (OMW) are a potential source of biophenols, but they have a complex composition with many unknown phenolics. The analysis of purified methanol extracts from two Portuguese OMW by electrospray mass spectrometry in the negative mode showed [M-H]- ions at m/z 539 and m/z 523, corresponding respectively to oleuropein and ligstroside isomers which contain the glucose unit linked to its aromatic moiety. Also, the fragmentation pathway of the [M-H]- ions at m/z 863, 685 and 847 indicated the presence of a diglucoside derivative of the oleuropein isomer and of mono- and diglucosides of the ligstroside isomer, respectively. Moreover, the two OMW samples contained an elenoic derivative of the ion at m/z 685 and a degradation product (m/z 453) of the [M-H]- ion at m/z 523. Future studies focusing on the abundance of these compounds on OMW, as well as their bioactivities, will determine their possible industrial exploitation.

Application of Fourier transform infrared spectroscopy and orthogonal projections to latent structures/partial least squares regression for estimation of procyanidins average degree of polymerisation.

Fourier transform infrared (FTIR) spectroscopy has being emphasised as a widespread technique in the quick assess of food components. In this work, procyanidins were extracted with methanol and acetone/water from the seeds of white and red grape varieties. A fractionation by graded methanol/chloroform precipitations allowed to obtain 26 samples that were characterised using thiolysis as pre-treatment followed by HPLC-UV and MS detection. The average degree of polymerisation (DPn) of the procyanidins in the samples ranged from 2 to 11 flavan-3-ol residues. FTIR spectroscopy within the wavenumbers region of 1800-700 cm(-1) allowed to build a partial least squares (PLS1) regression model with 8 latent variables (LVs) for the estimation of the DPn, giving a RMSECV of 11.7%, with a R(2) of 0.91 and a RMSEP of 2.58. The application of orthogonal projection to latent structures (O-PLS1) clarifies the interpretation of the regression model vectors. Moreover, the O-PLS procedure has removed 88% of non-correlated variations with the DPn, allowing to relate the increase of the absorbance peaks at 1203 and 1099 cm(-1) with the increase of the DPn due to the higher proportion of substitutions in the aromatic ring of the polymerised procyanidin molecules.

Antioxidant Capacities of Flavones and Benefits in Oxidative-Stress Related Diseases

Flavonoids, a group of secondary metabolites widely distributed in the plant kingdom, have been acknowledged for their interesting medicinal properties. Among them, natural flavones, as well as some of their synthetic derivatives, have been shown to exhibit several biological activities, including antioxidant, anti-inflammatory, antitumor, anti-allergic, neuroprotective, cardioprotective and antimicrobial. The antioxidant properties of flavones allow them to demonstrate potential application as preventive and attenuating agents in oxidative stress, i.e., a biological condition that is closely associated to aging processes and to several diseases. Some flavones interfere in distinct oxidative-stress related events by directly reducing the levels of intracellular free radicals (hydroxyl, superoxide and nitric oxide) and/or of reactive species (e.g. hydrogen peroxide, peroxynitrite and hypochlorous acid) thus preventing their amplification and the consequent damage of other biomolecules such as lipids, proteins and DNA. Flavones can also hinder the activity of central free radical-producing enzymes, such as xanthine oxidase and nicotinamide adenine dinucleotide phosphate oxidase (NADPH-oxidase) or inducible nitric oxide synthase (iNOS) and can even modulate the intracellular levels of oxidant and/or antioxidant enzymes. The evaluation of flavones antioxidant ability has been extensively determined in chemical or biological in vitro models, but in vivo therapy with individual flavones or with flavones-enriched extracts has also been reported. The present manuscript revises relevant studies focusing the preventive effects of flavones on stress-related diseases, namely the neurological and cardiovascular diseases and diabetes and its associated complications.

Overview on Mentha and Thymus Polyphenols

Mentha and Thymus are important genera of the Lamiaceae family widely distributed in the entire World and commonly used in traditional medicine. Indeed, many species of the two genera have been credited with a large list of health-benefit effects, including antioxidant, anti-inflammatory, antimicrobial, analgesic, neuroprotective and anticarcino- genic. In turn, these properties have been associated to the polyphenolic composition of the plants. The present review summarizes the phenolic constituents found in Mentha and Thymus genera, as well as the main methods applied in their extraction, purification and identification. Reported species of Mentha and Thymus usually comprise derivatives of caffeic acid and distinct glycosidic forms of the flavonoids luteolin, apigenin, eriodictyol and naringenin. At present, the phenolic composition of many relevant plants of Mentha and Thymus is still unknown and thus, more studies are required for the adequate phenolic characterization of these two genera. In this context, the present implementation of faster and reliable analytical methodologies, as e.g. the chromatographic techniques hyphenated with mass spectrometry, will surely be an enormous tool in the upgrading of the missing information.