Interferences of surfactants during determination of chlorophenols by acetylation coupled to headspace-solid phase microextraction-gas chromatography with an electron capture detector.

The micellar extraction (ME) of chlorophenols (CPs) from solid samples is rapid and avoids the use of organic solvents. The cationic surfactant cetyltrimethylammonium bromide (CTAB) is a powerful agent for ME of CPs. However, CTAB will be an important interference when the extracts are subsequently analyzed by direct solid phase microextraction (SPME) without a previous derivatization step. Therefore, CTAB is usually replaced by the nonionic surfactant polyoxyethylene-10-lauryl ether (POLE), which causes less interference but is less efficient in extracting CPs. In order to assess to what extent the derivatization of CPs with headspace (HS) sampling would be able to reduce surfactant interferences, CPs were measured in the presence of CTAB or POLE using acetylation and HS-SPME coupled to GC with an electron capture detector. Experiments were carried out both in water and in a 0.1 g/mL solution of NaCl (salting out agent). Acetylation and HS-SPME strongly decreased the interference of CTAB for CP determination. Therefore, the application of CTAB followed by SPME determination for an efficient ME of CPs from solid samples should be reconsidered since using CTAB with HS-SPME after acetylation proved to be a potential tool for CP determination in those matrixes after method optimization.

Trace metal concentration in a temperate freshwater reservoir seasonally subjected to blooms of toxin-producing cyanobacteria.

In situ interactions between cyanobacteria and metals were studied at Torrão reservoir (Tâmega River, North Portugal). The metal content of water and sediments from the reservoir was monitored monthly at Marco de Canaveses (seasonally subjected to toxic blooms of Microcystis aeruginosa) and upstream at Amarante (no blooms recorded), for 16 months. During the 16 months of the study period, M. aeruginosa bloomed twice at Marco de Canaveses, firstly forming a scum, and later with colonies scattered throughout the reservoir. Metals Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn were analysed in the sediment and in the water column. Cu-binding ligands in water were also determined. When no blooms were taking place, average metal levels for water and sediment were not statistically different at both locations. Therefore, it was considered that the absence of cyanobacteria blooms at Amarante was not due to differences in metal content. When blooms were taking place at Marco de Canaveses, a significant increase of metal levels in the sediment occurred simultaneously. Sediment quality guidelines showed that during this period, Cu and Pb concentrations (32.3 and 43.2 mg kg(-1), respectively) were potentially toxic. However, quantification of the exchangeable metal fraction indicated that these metals were probably not bioavailable. Concentration of Cu-binding ligands in water was higher during the blooms, indicating that cyanobacteria are capable of changing the metal speciation in situ in a reservoir.

Antioxidant response of Phragmites australis to Cu and Cd contamination.

Metals are known to induce oxidative stress in plant cells. Antioxidant thiolic compounds are known to play an important role in plants׳ defence mechanisms against metal toxicity but, regarding salt marsh plants, their role is still very poorly understood. In this work, the involvement of non-protein thiols (NPT), such as cysteine (Cys), reduced glutathione (GSH), oxidised glutathione (GSSG) and total acid-soluble SH compounds (total thiols), in the tolerance mechanisms of the marsh plant Phragmites australis against Cu and Cd toxicity was assessed. Specimens of this plant, freshly harvested in an estuarine salt marsh, were exposed, for 7 days, to rhizosediment soaked with the respective elutriate contaminated with Cu (0, 10 and 100 mg/L) or Cd (0, 1, 10 mg/L). In terms of NPT production, Cu and Cd contamination induced different responses in P. australis. The content of Cys increased in plant tissue after plant exposure to Cu, whereas Cd contamination led to a decrease in GSSG levels. In general, metal contamination did not cause a significant variation on GSH levels. Both metals influenced, to some extent, the production of other thiolic compounds. Despite the accumulation of considerable amounts of Cu and Cd in belowground tissues, no visible toxicity signs were observed. So, antioxidant thiolic compounds were probably involved in the mechanisms used by P. australis to alleviate metal toxicity. As P. australis is considered suitable for phytostabilising metal-contaminated sediments, understanding its tolerance mechanisms to toxic metals is important to optimise the conditions for applying this plant in phytoremediation procedures.

Stability of chlorophenols and their acetylated derivatives in water: sample storage procedures.

The determination of chlorophenols (CPs) in water samples is a subject of increasing interest. Reduction of sample storage space and the stability of CPs when present at very low levels are still problems that deserve research. The stability of CPs at ng/L levels at different temperatures and in the presence or absence of sodium carbonate and acetic anhydride was studied for up to 39 days. Stable and reproducible CP concentrations for about a month of storage in both river and wastewater were achieved in two storage conditions as follows: at -18 degrees C with addition of 10% sodium chloride; and at 4 degrees C with addition of both 10% sodium chloride and 10 mglmL sodium carbonate. These sample treatments are good alternatives to the immobilization of CPs on SPE cartridges in terms of both analyte stability and saving of storage space.

Simultaneous determination of several veterinary pharmaceuticals in effluents from urban, livestock and slaughterhouse wastewater treatment plants using a simple chromatographic method.

Minocycline, oxytetracycline, tetracycline, enrofloxacin and ceftiofur, commonly used veterinary pharmaceuticals, were searched in four urban, two livestock and two slaughterhouse effluents from wastewater treatment plants (WWTPs) in the north of Portugal. A simple method that includes solid-phase extraction followed with analysis by high-performance liquid chromatography with diode array detector was established and applied to the simultaneous determination of the five pharmaceuticals in WWTP effluents. This method, which is expeditious, inexpensive and available in most laboratories, showed to be useful for screening for problematic levels of drugs in WWTP effluents. It is known that several livestock and slaughterhouse effluents (pre-treated or treated) are discharged to the urban network before discharge into the environment. The presence of these drugs in such effluents can constitute a significant environmental problem that should be addressed, by the monitoring of these drugs and by implementation of methodologies that contribute to their decrease/elimination from wastewaters. Minocycline (≤6 µg L(-1)), oxytetracycline (≤7 µg L(-1)), tetracycline (≤6 µg L(-1)) and enrofloxacin (<2 µg L(-1)) could be detected and/or quantified in three urban effluents. Detectable levels of enrofloxacin (<2 µg L(-1)) and quantifiable levels of tetracycline (≤15 µg L(-1)) were found in the slaughterhouse effluents.

A headspace SPME-GC-ECD method suitable for determination of chlorophenols in water samples.

A headspace solid phase microextraction coupled to gas chromatography with electron capture detector (HS-SPME-GC-ECD) method was optimized for the determination of seven chlorophenols (CPs) with different levels of chlorination. This is the first time that HS-SPME-GC-ECD with acetylation of the analytes is used for the simultaneous determination of CPs in water samples. The influence of fibre type, derivatization conditions, salt addition, temperature and time of extraction and temperature of desorption was checked. Possible sources of contamination and analyte losses were considered. The best results were obtained with the polydimethylsiloxane/divinylbenzene fibre, derivatization by acetylation using 100 µL of acetic anhydride and 0.1 g of anhydrous sodium carbonate per 10 mL of sample, salt addition of 100 g L(-1) sodium chloride, extraction at 70 °C for 60 min and desorption in the GC injector at 260 °C for 6 min. The limits of detection (LOD) for monochlorophenols were 12 and 122 ng L(-1) for 2-chlorophenol and 4-chlorophenol, respectively. For polychlorinated CPs, the LODs were lower than 6 ng L(-1), values similar to the existing methods that use SPME with derivatization for CPs determination in water samples. The method is suitable for the determination of CPs in most environmental aqueous samples. Repeatability and reproducibility were less than 16.8% and 11.7%, respectively. The optimized method was successfully applied for the analysis of waters with complex matrices such as river and estuarine water samples.

Effects of minocycline and its degradation products on the growth of Microcystis aeruginosa.

This work aimed to investigate the influence of Microcystis aeruginosa on the rate of minocycline (MNC) removal (abiotic degradation, physical binding or chemical transformation by cells) and the toxicity of MNC and its degradation products to the cyanobacterium. Most of the work was carried out in culture media in the presence or in the absence of M. aeruginosa. The rate of the MNC abiotic degradation in culture media strongly decreased with the increase of the MNC initial concentration. The exudates did not influence the rate of MNC degradation in the media. For concentrations ≥0.48 µM, the rate of the MNC removal from the media was faster in the presence of M. aeruginosa. Biotic MNC removal occurs by both physical binding by the cells (10%) and chemical transformations. EC(50) and EC(20) for MNC on the growth of M. aeruginosa were 0.92 and 0.13 µM, respectively. Interestingly, MNC degradation products might promote M. aeruginosa growth.

Allelopathy in freshwater cyanobacteria.

Freshwater cyanobacteria produce several bioactive secondary metabolites with diverse chemical structure, which may achieve high concentrations in the aquatic medium when cyanobacterial blooms occur. Some of the compounds released by cyanobacteria have allelopathic properties, influencing the biological processes of other phytoplankton or aquatic plants. These kinds of interactions are more easily detectable under laboratory studies; however their ecological relevance is often debated. Recent research has discovered new allelopathic properties in some cyanobacteria species, new allelochemicals and elucidated some of the allelopathic mechanisms. Ecosystem-level approaches have shed some light on the factors that influence allelopathic interactions, as well as how cyanobacteria may be able to modulate their surrounding environment by means of allelochemical release. Nevertheless, the role of allelopathy in cyanobacteria ecology is still not well understood, and its clarification should benefit from carefully designed field studies, chemical characterization of allelochemicals and new methodological approaches at the "omics" level.

Butyltin levels in several Portuguese coastal areas.

This work aimed to report present levels (2007-2008 sampling) of tri- (TBT), di- (DBT), and monobutyltin (MBT) in surface sediments from 11 Portuguese coastal sites and discuss the evolution of BTs contamination in the last two decades. All the samples revealed quantifiable values of TBT, DBT, and MBT with total butyltin concentrations between 1 and 565 ng/g (of Sn in dry sediment). Maximum level of TBT, 66 ng/g, was observed in Sado estuary, at Lisnave site, in the proximity of a big shipyard. MBT decreased site by site by the same order as DBT and TBT did, but its concentrations were much higher in many cases, denoting that TBT contamination was much higher in the past. A comparison with the available previous data confirmed a marked decrease of TBT contamination all over the last years, indicating that the main sources of TBT in Portuguese coastwise stopped effectively.

Application of SPME to the determination of alkylphenols and bisphenol A in cyanobacteria culture media.

In order to survey the influence of estrogenic compounds on cyanobacteria, solid-phase microextraction (SPME) with a carbowax-divinylbenzene fibre was used for the determination of tert-octylphenol (tert-OP), n-nonylphenol (n-NP), technical nonylphenol (tech-NP) and bisphenol A (BPA) in cyanobacteria culture media by gas chromatography with flame ionization detection. Determinations were carried out without derivatization in deionized water and filtered culture media. A comparison between f2 and Fraquil culture media was performed, which showed that only f2 allowed quantitative recoveries. Headspace SPME with salting out, requiring only 10 mL of sample, was suitable for tert-OP, n-NP, and tech-NP determination with limits of detection (LOD) of <0.05 microg L(-1). For BPA, direct immersion SPME could provide a LOD of 1 microg L(-1). Automated sampling allowed reproducible extraction. No exudate substances overlapped with the studied compounds during the chromatographic separation and no matrix effects were observed. Ecotoxicity tests can be performed by single spiking of tert-OP and tech-NP and multiple spiking of n-NP due to its lower stability.

Influence of a salt marsh plant (Halimione portulacoides) on the concentrations and potential mobility of metals in sediments.

Influence of Halimione portulacoides, commonly found in temperate salt marshes, on sediment metal contents, speciation and potential mobility in case of sediment re-suspension was evaluated. Both colonized and non-colonized sediments were studied for total Cd, Cu, Pb and Zn contents and metal fraction exchangeable to water collected in situ. Sediment elutriates, prepared with water collected from each site, were used to simulate a sediment re-suspension phenomenon. As the characteristics and degree of contamination of sediments may influence system behaviour, salt marshes of two Portuguese estuaries, Cavado (NW coast) and Sado (SW coast), were studied. Cu, Pb and Zn contents higher than ERL (quality guideline, effect range-low) were observed, indicating potential risks for living organisms. Strong Cu-complexing organic ligands, also determined in both water and elutriates, were higher in rhizosediment elutriates, at concentrations similar, or even higher, to those of Cu. Such ligands condition metals speciation in the water column and probably also metal bioavailability. From rhizosediment significant amounts of Cu and Zn were transferred to the aqueous phase, concentrations 2-8 times higher than concentrations present in water. In contrast, elutriates of non-colonized sediment removed metals from water, Cu and Zn levels in elutriates being 2-6 times lower than initial ones. Cd and Pb levels in water and elutriates were not measurable in most cases. Results clearly indicate that metals potential solubility in the rhizosphere of plants was markedly higher than that in the surrounding sediment. The obtained results indicated that H. portulacoides presence (and probably other salt marsh plants) may cause a marked increase in metals concentrations in dissolved phase (pore water or even water column if rhizosediment is re-suspended). As salt marsh plants may be abundant in temperate and subtropical estuaries and costal lagoons, this phenomenon should not be disregard in future studies towards the sustainable management of such environments.

Copper, nickel and lead in lichen and tree bark transplants over different periods of time.

This work aimed at comparing the dynamics of atmospheric metal accumulation by the lichen Flavoparmelia caperata and bark of Platanus hybrida over different periods of time. Transplants were exposed in three Portuguese coastal cities. Samples were retrieved (1) every 2 months (discontinuous exposure), or (2) after 2-, 4-, 6-, 8- and 10-month periods (continuous exposure), and analysed for Cu, Ni and Pb. Airborne accumulation of metals was essentially independent of climatic factors. For both biomonitors [Pb]>[Ni]>[Cu] but Pb was the only element for which a consistent pattern of accumulation was observed, with the bark outperforming the lichen. The longest exposure periods hardly ever corresponded to the highest accumulation. This might have been partly because the biomonitors bound and released metals throughout the exposure, each with its own dynamics of accumulation, but both according to the environmental metal availability.

Exudates of different marine algae promote growth and mediate trace metal binding in Phaeodactylum tricornutum.

Phaeodactylum tricornutum was grown in filtered natural seawater enriched with nitrate, phosphate, and silicate only (control) or with exudates from itself, from Emiliania huxleyi (a coccolithophore micro-alga), Porphyra spp. (a red macro-alga) or Enteromorpha spp. (a green macro-alga). Cathodic (and anodic) stripping voltammetry (C(A)SV) were used to determine the concentrations of trace metals, both in the medium and in the algae, as well as total Cu-complexing organic ligands in the medium and, among these, some thiols (compounds identified as cysteine- or as glutathione by CSV). Exudates of different marine micro- and macro-algae could cause allelopathic effects in P. tricornutum cultures. Cell yield of P. tricornutum was increasingly promoted by exudates of E. huxleyi >Porphyra >Enteromorpha. Although exudates strongly complex Cu (and probably other metals), their presence promoted Cu uptake. Significant changes of Ni, Cd, Fe, Zn and Mn uptake by P. tricornutum were also observed in the presence of exudates of different algal species. In addition, both intensity of production and nature of exudates released by P. tricornutum were markedly influenced by the presence of exudates of other algae, the allelopathic effects being very specific (variable from one species to another). Allelopathy will probably also occur in the aquatic environment, although to a lesser extent than in cultures, particularly during bloom events and may have effects on both chemical speciation and bioavailability of chemicals to phytoplanktonic species. Such changes might cause the predominance of some species over other species. Therefore, in future in vitro culture studies with the purpose of using them as models of the real environment, more attention should be paid to the role of algal exudates, in order to improve the environmental relevance and significance of the results.

Can PAHs influence Cu accumulation by salt marsh plants?

The presence of polycyclic aromatic hydrocarbons (PAHs) may change the mechanisms of metal uptake, thus influencing kinetics and extent of metal phytoextraction. Studies on the subject are scarce, particularly for salt marsh plants. The aim of this work was to investigate the effect of PAHs on the uptake of Cu by Halimione portulacoides, a plant commonly found in salt marshes. Experiments were carried out in the laboratory, either in hydroponics (sediment elutriate) or in sediment soaked in elutriate, which were prepared with sediment and water from a salt marsh of the Cavado river estuary (NW Portugal). Groups of H. portulacoides (grown in a greenhouse) were exposed to those media during six days. Cu2+ (as Cu(NO3)2), 10(2) and 10(4) microg l(-1), was added to the media as well as 1.6 microg l(-1) of the sixteen EPA priority PAHs (0.1 microg l(-1) of each PAHs). Cu was assayed in solutions, sediments and different plant tissues before and after experiments. After exposure, photosynthetic efficiency and levels of chlorophylls were also measured, indicating that plant stress indicators were identical in all plants independently of the media to which the plants were exposed. PAHs influenced both the soluble Cu fraction and Cu uptake by plants. The amounts of metal accumulated in both roots and stems were significantly higher when the 10(4) microg l(-1) of Cu enriched elutriate was amended with PAHs. Thus, results suggest that PAHs may modify Cu solubility, the Cu sorption by plants and/or the passive penetration of Cu into the root cells. Therefore, the combined effects of different types of pollutants should be taken in consideration when studying the remediation potential of plants, namely in terms of phytoextraction.

Comparison of the role of the sea club-rush Scirpus maritimus and the sea rush Juncus maritimus in terms of concentration, speciation and bioaccumulation of metals in the estuarine sediment.

The capability of Scirpus maritimus and of Juncus maritimus to accumulate metals and the role of each plant on the physico-chemical composition of sediments, from Douro river estuary (NW Portugal), were investigated through a year of plants' life. The contents of Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn were determined (by atomic absorption spectrophotometry) in sediment, rhizosediments (those in contact with each plant roots and rhizomes) and different tissues of S. maritimus and of J. maritimus. Both plants influenced the sediment composition (concentrating metals around its roots) and were able to bioaccumulate Cd, in spite of some seasonal variations in the metal levels in the system (both in rhizosediments and plants' roots). Therefore, both plants showed to have potential for Cd phytostabilization. S. maritimus could also concentrate Pb in its roots, probably by sorption on the root surface in (hydr)oxide forms, therefore, denoting capability for Pb phytostabilization.

Variability of metal contents in the sea rush Juncus maritimus-estuarine sediment system through one year of plant's life.

Metal accumulation by Juncus maritimus and the effect this plant might have on the physico-chemical composition of sediments, from Douro river estuary (NW Portugal), were investigated through one year of plant's life. The contents of Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn were determined (by atomic absorption spectrophotometry) in sediments, rhizosediments (those among plant roots and rhizomes) and different tissues of J. maritimus. Two estuarine sites with different characteristics were studied, one a sandy sediment with relatively low organic matter content, the other a muddy site. The role of the plant seemed to be markedly dependent on the sediment composition. At the sandy site, significant variations throughout the year (a decrease from winter to summer and an increase in autumn) either in rhizosediment and sediment or in J. maritimus roots were only observed in Cd and Cu levels. These variations were the same in both compartments, probably caused by tidal water movements and the activity of the plant. At the muddy site, variations throughout the year were observed for all studied elements but Pb. Changes in the Cd and Zn contents of roots were opposite to those in the rhizosediment (uptake of Cd and Zn by J. maritimus roots in summer may have caused a decrease in rhizosediment levels). Fe and Mn showed similar patterns of variation in both roots and rhizosediment (contents significantly increasing in summer). Higher contents of Fe and Mn in roots in summer could be related to changes in mass, higher availability of the elements, or accumulation at the surface of the root in (hydr)oxides forms. For the remaining elements no significant correlations between rhizosediment and roots variations were observed, indicating that their uptake and release by roots were not based on single exchanges with the sediment. J. maritimus showed capacity to accumulate Cu, Zn and particularly Cd, thus appearing to be useful for the phytostabilization of these metals.

Spatial and seasonal variations of the macrobenthic community and metal contamination in the Douro estuary (Portugal).

The macrobenthic community structure and characteristics of sediments were studied in samples collected seasonally at five stations in the lower mesotidal Douro estuary, Portugal. Sediment characterisation included grain size distribution, total-recoverable metals (Al, Fe, Mn, Zn, Cu, Cr, Pb, Ni and Cd), acid-volatile sulphides (AVS) and simultaneously extracted metals (SEM) and total organic matter contents. Spatial variation surpassed temporal variation both in terms of environmental parameters and community structure. Clear anthropogenic contamination in terms of Zn, Cr, Cu and Pb, and a positive SEM-AVS, indicating the potential bioavailability of these metals, were detected on the north bank of the estuary. Multivariate analysis revealed that in addition to natural sediment characteristics, the anthropogenic metal contamination was influencing the macrobenthic community structure. In fact, greater concentrations of Zn, Cr, Cu and Pb were associated with lower number of species and/or diversity. Finally, the dry weight-normalised metal concentrations appeared as a more useful tool than the SEM-AVS model in predicting disturbance of the macrobenthic community in the studied area.

The role of a salt marsh plant on trace metal bioavailability in sediments. Estimation by different chemical approaches.

GOAL, SCOPE AND BACKGROUND: The presence or absence of vegetation can condition sediment characteristics. The main aim of this work was to investigate the influence of the sea rush Juncus maritimus on metal (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) availability to organisms living on or in estuarine sediments, from Douro River (NW Portugal), by comparing the characteristics and chemical behaviour of rhizosediments (collected within the plant assemblage) and those of sediment (collected around the plant). In order to evaluate whether and how sediment characteristics condition the role of plants on metal availability, sandy and muddy sediments colonised by J. maritimus were studied in parallel. METHODS: Metal availability was estimated by enzymatic digestion with pepsin (ED), which may provide an estimate of metal availability to organisms living at estuarine sediments. Nevertheless, since no consensus exists yet on the most suitable methodologies to estimate metal bioavailability in sediments, two more conventional approaches, BCR sequential extraction (SE) and AVS/ SEM model, were also used, in parallel, and the information these approaches provided was compared with that provided by ED. Total-recoverable metal contents were determined by atomic absorption spectrophotometry after sediment digestion using a high-pressure microwave system. RESULTS AND DISCUSSION: Plants could concentrate metals around its roots and rhizomes. In addition, they were capable of oxidizing (release of oxygen by the roots) the anaerobic medium surrounding their roots in muddy sediment (reducing AVS). As sulphide oxidation renders metals (Cd, Cu, Ni, Pb and Zn) into more soluble forms, according to the AVS/SEM model, metals from muddy sites would be more available in rhizosediment than in sediment. The SE approach led to a similar conclusion. Nevertheless, the results provided by ED pointed at opposite conclusion, particularly for Cd and Zn, indicating less availability at rhizosediments than in the surrounding sediment. ED results were interpreted as a consequence of an enrichment of the rhizosediment in organic ligands exuded by the roots or liberated by dead plants. The effect of complexation of metals by organic compounds, which ED could not decompose/dissolve, seemed to overcome that caused by sediment oxidation. In general, a comparison of the informa tion about metal availability by ED, SE, AVS/SEM, showed that it did not always match and in few cases it was even contradictory. CONCLUSION AND OUTLOOK: Therefore, a thorough evaluation of the metal availability in sediments requires a combination of different chemical approaches, so as to take into consideration differences in ways of organism exposure (interstitial water and/or ingestion of sediment particles).

N-(2-Carboxyethyl)chitosans: regioselective synthesis, characterisation and protolytic equilibria.

N-(2-Carboxyethyl)chitosans were obtained by reaction of low molecular weight chitosan with a low degree of acetylation and 3-halopropionic acids under mild alkaline media (pH 8-9, NaHCO3) at 60 degrees C. The chemical structure of the derivatives obtained was determined by 1H and 13C NMR spectroscopies. It was found that alkylation of chitosan by 3-halopropionic acids proceeds exclusively at the amino groups. The products obtained are described in terms of their degrees of carboxyethylation and ratio of mono-, di-substitution and free amine content. The protonation constants of amino and carboxylate groups of a series of N-(2-carboxyethyl)chitosans were determined by pH-titration at ionic strength 0.1 M KNO3 and 25 degrees C.

Lead contamination in Portuguese red wines from the Douro region: from the vineyard to the final product.

To quantify lead contamination in wines and to try to identify major lead sources, two winemaking processes were followed during one annual cycle of wine production. Two vineyards from the Douro Portuguese region and two types of wine, one red table wine, which has been produced in a very modern winery, and one red fortified wine (similar to Port), which has been produced by a traditional vinification process, were selected for this study. Aerosols from the vineyards atmosphere, vineyard soil, vine leaves, grapes, and samples from the intermediary and final wine product were collected. Suitable pretreatments, namely, high-pressure microwave assisted digestion (soil, leaves, and grapes) and UV-irradiation (grape juices and samples from the different steps of the vinification processes), were used. The samples were analyzed in terms of lead total concentration and respective isotope ratios by using inductively coupled plasma mass spectrometry and atomic absorption spectrophotometry with electrothermal atomization. It was observed that the major sources of lead were in the vinification system, the more traditional one introducing more lead than the modern one. For the fortified wine, the lead concentration increased from 4.7 microg L(-1), in the grape juice, to 17.2 microg L(-1), in the final product, while for the table wine the increase was from 4.1 to 13.1 microg L(-)(1). Therefore, only about 1/4 (fortified wine) and 1/3 (table wine) of the lead total content of the final products came from soil and atmospheric deposition. Therefore, it is expected that marked reductions of the lead content in the wines would occur if the sources of lead were removed from the tubes and containers used in the vinification system, particularly by using welding alloys and small fittings free of lead. The lead levels in the vine leaves (global mean of 0.43 microg g(dry leave)(-1)) and grapes (global mean of 35 ng g(dry grape)(-1)) were similar in both vineyards.