Chemometric-assisted construction of a biosensing device to measure chlorogenic acid content in brewed coffee beverages to discriminate quality.

In this work we propose the use of statistical mixture design in the construction of a biosensor device based on graphite oxide, platinum nanoparticles and biomaterials obtained from Botryosphaeria rhodina MAMB-05. The biosensor was characterized by electrochemical impedance spectroscopy. Under optimized experimental parameters by factorial design, the biosensor was applied to the voltammetric determination of chlorogenic acid (CGA) measured as 5-O-caffeoylquinic acid (5-CQA). The biosensor response was linear (R2 = 0.998) for 5-CQA in the concentration range 0.56-7.3 µmol L-1, with limit of detection and quantification of 0.18 and 0.59 µmol L-1, respectively. The new biosensing device was applied to quality control analysis based upon the determination of CGA content in specialty and traditional coffee beverages. The results indicated that specialty coffee had a significantly higher content of CGA. Principal component analysis of the voltammetric fingerprint of brewed coffees revealed that the laccase-based biosensor can be used for their discrimination.

Factorial design fingerprint discrimination of Coffea arabica beans under elevated carbon dioxide and limited water conditions.

Coffee is one of the most important commodities, showing sensitivity to environmental variations. The main effects and their interaction for two levels of atmospheric CO2 concentrations and two water regimes of a factorial design were investigated for the metabolic profiles of Coffea arabica raw beans using UV fingerprint analysis from a mixture design. UV fingerprint results obtained from pure ethanol and binary ethanol-dichloromethane mixtures showed the largest metabolic discriminations between CO2 levels and their extracts were investigated in detail. The biosynthesis of major metabolites, chlorogenic acids, cafestol, kahweol and caffeine were altered owing to environmental conditions. Higher amounts of chlorogenic acids and kahweol were observed in beans from unirrigated plants grown with enriched CO2 and irrigated ones at the current CO2 level. Water availability and CO2 concentration interaction affects the metabolite amounts. Besides a significant CO2 atmospheric effect water availability was a limiting factor for metabolite content only at current CO2 level, suggesting the successful metabolic coping of CO2 enriched Arabic coffee beans suffering future droughts.

Seasonal changes and solvent effects on fractionated functional food component yields from Mikania laevigata leaves.

Statistical mixture design extraction and fractionation of Mikania plant samples were carried out to quantitatively study seasonal and solvent composition effects as well as their interactions on secondary metabolites. The mixture design consisted of ethanol, acetone, dichloromethane and chloroform solvents and their binary, ternary and quaternary mixtures. Yields were measured for the crude extract and its neutral, organic, basic, polar and fiber fractions obtained with each solvent composition from samples harvested during 2010. Two-way ANOVA found statistically significant seasonal and solvent effects for the crude extract and all the fractions except for the polar fraction solvent. The best solvent extractor depends the harvest season. High crude, organic and polar fraction yields in the summer are correlated with the coumarin UV-vis absorbance at 274 nm. Crude yields of mixtures containing ethanol are correlated with the coumarin absorbances in summer, winter and spring whereas mixtures without ethanol show no significant correlation.

Sequential mixture design optimization for divergent metabolite analysis: Enriched carbon dioxide effects on Coffea arabica L. leaves and buds.

The first metabolic study of the impact of elevated CO2 (590 µL CO2 L-1) levels on the leaves and buds of Coffea arabica L. plants is reported. A novel sequential statistical mixture design strategy allowed optimization of both the extraction and mobile phase solvent systems to increase differences detected in metabolites of Coffea arabica L. plants and buds. Factor analysis showed that the 227 and 273 nm bands of the 1:1:1 ternary ethyl ether - dichloromethane - methanol mixture spectra resulted in discrimination of elevated CO2 extract samples from those obtained from leaves grown in a current level CO2 atmosphere (390 µL CO2 L-1) of leaf sample extracts. DAD-HPLC spectral peak evidence showed a 32% increase in absorbance of the 273 band for the enriched CO2 leaf extracts. This band has been assigned to caffeine-like substances and confirmed by the mass spectral signal at m/z 195 ([M + H]+). No enrichment band increases were found for kahweol, kaempferol and quercetin that had presence confirmed by mass spectral analysis. No epigenetic effect of this metabolic profile was found in new leaves after the addition of CO2 stopped. Enriched CO2 perturbation of the bud metabolite were much smaller than for the leaf samples. Absorbance increases in the 228 nm and decreases in the 235 nm bands play a prominent role in the discrimination of enriched CO2 buds from the controls in the pure dichloromethane extracting solvent. This global metabolome strategy allows the monitoring of chemical groups of plants susceptible to environmental changes as well as elucidate metabolic variations in complex matrices of biochemical responses.

Antibacterial activity of Limonium brasiliense (Baicuru) against multidrug-resistant bacteria using a statistical mixture design.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze (Plumbaginaceae) is commonly known as "baicuru" or "guaicuru" and preparations of its dried rhizomes have been popularly used in the treatment of premenstrual syndrome and menstrual disorder, and as an antiseptic in genito-urinary infections. This study evaluated the potential antibacterial activity of rhizome extracts against multidrug-resistant bacterial strains using statistical mixture design. MATERIALS AND METHODS: The statistical design of four components (water, methanol, acetone, and ethanol) produced 15 different extracts and also a confirmatory experiment, which was performed using water:acetone (3:7, v/v). The crude extracts and their ethyl-acetate fractions were tested against vancomycin-resistant Enterococcus faecium (VREfm), methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae, all of which have been implicated in hospital and community-acquired infections. The dry residue, total polyphenol, gallocatechin and epigallocatechin contents of the extracts were also tested and statistical analysis was applied in order to define the fit models to predict the result of each parameter for any mixture of components. The principal component and hierarchical clustering analyses (PCA and HCA) of chromatographic data, as well as mass spectrometry (MS) analysis were performanced to determine the main compounds present in the extracts. RESULTS: The Gram-positive bacteria were susceptible to inhibition of bacterial growth, in special the ethyl-acetate fraction of ternary extracts from water:acetone:ethanol and methanol:acetone:ethanol against, respectively, VREfm (MIC=19µg/mL) and MRSA (MIC=39µg/mL). On the other hand, moderate activity of the ethyl-acetate fractions from primary (except water), secondary and ternary extracts (MIC=625µg/mL) was noted against KPC. The quadratic and special cubic models were significant for polyphenols and gallocatechin contents, respectively. Fit models to dry residue and epigallocatechin contents were not possible. PCA and HCA of the chromatographic fingerprints were disturbed by displacement retention time of some peaks, but the ultraviolet spectra indicated the homogeneous presence of flavan-3-ols characteristic of tannins. The MS confirmed the presence of gallic acid, gallocatechin, and epigallocatechin in extracts, and suggested the presence of monomers and dimers of B- and A-type prodelphinidins gallate, as well as a methyl gallate. CONCLUSION: Our results showed the antibacterial potential of L. brasiliense extracts against multidrug-resistant Gram-positive bacteria, such as VREfm and MRSA. The statistical design was a important tool to evaluate the biological activity by optimized form. The presence of some phenolic compounds was also demonstrated in extracts.

Saliva as a potential tool for diagnosis of dry mouth including Sjögren's syndrome.

OBJECTIVES: Recently, the use of saliva as a diagnostic tool has gained considerable attention because it is non-invasive and easy to perform repeatedly. In this study, we focused on soluble molecules in saliva to establish a new diagnostic method for xerostomia. MATERIALS AND METHODS: Saliva was obtained from 90 patients with Sjögren's syndrome (SS), 22 patients with xerostomia associated with neurogenic/neuropsychiatric disorders and drugs (XND), 30 patients with radiation-induced xerostomia (RX), and 36 healthy controls. Concentrations of helper T (Th) cytokines in saliva were measured by flow cytometric analysis. Concentrations of secretory IgA (SIgA) and chromogranin A (CgA) were measured by ELISA. RESULTS: Unstimulated and stimulated whole saliva from patients with SS, XND, and RX was significantly reduced compared with controls. Th1 and Th2 cytokines from SS patients were significantly higher than controls. Furthermore, Th2 cytokines were closely associated with strong lymphocytic accumulation in salivary glands from SS patients, while Th1 and Th17 cytokines were negatively associated. SIgA levels were not significantly different between all patient groups and controls. CgA levels from XND patients were significantly higher than controls. CONCLUSIONS: The measurement of cytokines, CgA, and SIgA in saliva is suggested to be useful for the diagnosis of xerostomia and also to reveal disease status.

Evaluation of the photodynamic activity of xanthene dyes on Artemia salina described by chemometric approaches.

The development of drugs for photodynamic therapy (PDT) is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y) with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i) photosensitizers concentration, (ii) the LED irradiation time and (iii) the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 2³ full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

Close association between oral Candida species and oral mucosal disorders in patients with xerostomia.

OBJECTIVE: Heightened interest in oral health has lead to an increase in patients complaining of xerostomia, which is associated with various oral mucosal disorders. In this study, we investigated the relationship between Candida species and oral mucosal disorders in patients with xerostomia. SUBJECTS AND METHODS: We evaluated whole salivary flow rate and presence of oral mucosal disorders in 48 patients with xerostomia and 15 healthy controls. The number of Candida species was measured as colony-forming units after propagation on selective medium. Identification of Candida at the species level was carried out by polymerase chain reaction and restriction fragment length polymorphism analysis. We then examined the relationship between Candida species and oral mucosal symptoms. RESULTS: Compared with controls, patients with xerostomia exhibited significantly decreased whole salivary flow rate, increased rate of oral mucosal symptoms, and higher numbers of Candida. Salivary flow rate negatively correlated with the number Candida. Among patients with oral candidiasis, Candida albicans was isolated from the tongue mucosa and Candida glabrata was isolated from the angle of the mouth. CONCLUSION: These results suggest that particular Candida species are involved in the pathogenesis of oral mucosal disorders in patients with xerostomia.

Mixture designs for exploring class diversity and metabolite fingerprinting: an efficient column chromatographic strategy.

The effects of four solvents, hexane, dichloromethane, ethyl acetate, methanol, and their mixtures on the separation of metabolites in crude extracts of Erythrina speciosa Andrews leaves were investigated using two strategies for open column chromatography. The classical extraction procedure was compared with mobile phases prepared according to a mixture design in order to explore the effects of solvent interactions on metabolite separations. Principal component analysis was used to compare the UV spectra obtained from RP-HPLC-DAD and to estimate the number of independent factors contained in the chromatographic data of the extracts. The results showed that, in addition to solvent polarity, solvent mixtures play an important role in metabolite separation. When pure solvents are used, larger groups of similar spectra are observed in the factor analysis score graphs indicating the same or a limited number of metabolite classes. In contrast solvent mixtures produced score graphs with a larger number of clusters indicating greater metabolic diversity. Besides resulting in more peaks than the pure solvents the chromatographic data of the design mixtures resulted in larger numbers of significant principal components confirming the greater chemical diversity of their extracts. Thus, if the objective of an analysis is to obtain metabolites of the same class, one should use pure solvents. On the other hand, binary and ternary solvent mixtures are recommended for more efficient investigations of class diversity and richer metabolite fingerprints.

Behavioral effects of aqueous and dichloromethane extracts of Erythrina speciosa Andrews, Fabaceae, leaves in mice / Efeitos comportamentais do extrato aquoso bruto e fração diclorometano das folhas de Erythrina speciosa Andrews, Fabaceae, em camundongos

Erythrina speciosa Andrews, Fabaceae, is used in the South of Brazil as sedative and tranquilizers. In this study, behavioral effects of aqueous (AE) and dichloromethane (DCM) extracts of E. speciosa leaves were evaluated in male mice, as well as their lethal dose 50 percent (LD50). The extracts were administered by gavage in doses ranging from 50 to 400 mg/kg 1 h before the behavioral tests. AE decreased ambulation (50, 100 and 400 mg/kg) and rearing (50 and 400 mg/kg) in the open-field but did not influence elevated plus maze, rotarod and pentobarbital-induced sleep tests. No behavioral effects were observed after DCM administration. LD50 for both extracts were higher than 2000 mg/kg. The results showed that AE and DCM extracts of E. speciosa leaves do not produce anxiolytic effect in the elevated plus maze nor seems to depress the Central Nervous System. However, since serotonergic mechanisms may be involved in the pharmacological action of Erythrina plants and the elevated plus maze test is not adequate to evaluate serotonergic drugs, our results do not invalidate the use of this plant in folk medicine but suggest that the mechanism involved in a possible central action of Erythrina needs to be clarified.

Erythrina speciosa Andrews, Fabaceae, é usada na região Sul do Brasil como sedativa e tranquilizante. Neste estudo, uma possível ação central do extrato aquoso (EA) e fração diclorometano (DCM) das folhas da E. speciosa foi avaliada em camundongos machos submetidos a testes comportamentais 1 h após o tratamento (gavage). Também foi avaliada a dose letal 50 por cento (DL50) como indicativa da toxicidade aguda desta planta. O EA diminuiu a locomoção (50, 100 e 400 mg/kg) e o levantar (50 e 400 mg/kg) no teste de campo aberto mas não alterou nenhum dos comportamentos avaliados nos testes de labirinto em cruz elevado, rotarod e sono induzido. A DCM não alterou nenhum dos comportamentos avaliados. A DL50 de ambos os extratos foi estimada como sendo >2000 mg/kg. Os resultados sugerem ausência de efeito ansiolítico e depressor do Sistema Nervoso Central das folhas de E. speciosa. Entretanto, como mecanismos serotonérgicos podem estar envolvidos na ação farmacológica de plantas do gênero Erythrina e o teste de labirinto em cruz elevado não é adequado para avaliar o efeito de drogas serotonérgicas, nossos resultados não invalidam o uso desta planta na medicina popular, mas apontam a necessidade de se investigar o mecanismo de ação envolvido no possível efeito central de plantas do gênero Erythrina.

Antinociceptive and acute toxicity evaluation of Vernonia condensata Baker leaves extracted with different solvents and their mixtures.

Extract of Vernonia condensata (Asteraceae = Compositae) leaves has different uses in Brazilian folk medicine, which includes analgesic and antiinflamatory agent. The aim of this study was to apply a modified simplex-centroid mixture design to evaluate the best extractor system for the antinociceptive activity, evaluated by writhing test. Different solvents (acetone, dichloromethane, ethanol and ethyl acetate) as well as their binary, ternary and quaternary mixtures were used. For comparison, aqueous extract was also evaluated. LD50 was estimated and qualitative phytochemical screening, conducted. The extracts with antinociceptive activity were: aqueous, acetone, dicloromethane (DCM), ethanol (ETOH), acetone-DCM, acetone-ETOH, acetone-ethyl acetate, ETOH-ethyl acetate, acetone-DCM-ethyl acetate, acetone-ETOH-ethyl acetate and DCM-ETOH-ethyl acetate. The higher margin of safety (LD50/ED50) was for acetone > acetone-ETOH-ethyl acetate > aqueous > ETOH = acetone-ETOH > DCM > acetone-ethyl acetate > DCM-ETOH-ethyl acetate > acetone-DCM > acetone-DCM-ethyl acetate. Phytochemical screening showed that all the extracts contained alkaloids, phenolic compounds, flavonoids, tannins and saponins. In conclusion, the extractor system influences both the pharmacological activity and acute toxicity of leaves from V. condensata. Acetone and the ternary mixture, acetone-ETOH-ethyl acetate extracts showed higher margin of safety than aqueous extract.

Statistical mixture design--varimax factor optimization for selective compound extraction from plant material.

Varimax-transformed chromatograms of compounds extracted from Erythrina speciosa Andrews leaves by simplex centroid design mixtures of dichloromethane, hexane, ethanol and acetone are reported and compared with principal component results. Six varimax factors were investigated focusing on the three main groups of extracted compounds with retention times of 1.7, 3.1 and 6.6 min. Varimax models provide chromatographic loading profiles that are simpler than their principal component counterparts. Furthermore varimax score models in terms of extraction medium compositions are simpler to interpret. The first varimax model results in an optimum extraction mixture of 71% dichloromethane-29% acetone although substitution of acetone by ethanol results in an almost identical extraction profile. The second varimax score model predicts optimum extraction binary mixtures with hexane proportions ranging from 50 to 100% for complementary proportions of either acetone or ethanol. The third varimax factor provides a response surface that is very similar to the one found for the third PC. Confirmatory experiments were performed to validate the model predictions.

Mixture-mixture design for the fingerprint optimization of chromatographic mobile phases and extraction solutions for Camellia sinensis.

A composite simplex centroid-simplex centroid mixture design is proposed for simultaneously optimizing two mixture systems. The complementary model is formed by multiplying special cubic models for the two systems. The design was applied to the simultaneous optimization of both mobile phase chromatographic mixtures and extraction mixtures for the Camellia sinensis Chinese tea plant. The extraction mixtures investigated contained varying proportions of ethyl acetate, ethanol and dichloromethane while the mobile phase was made up of varying proportions of methanol, acetonitrile and a methanol-acetonitrile-water (MAW) 15%:15%:70% mixture. The experiments were block randomized corresponding to a split-plot error structure to minimize laboratory work and reduce environmental impact. Coefficients of an initial saturated model were obtained using Scheffe-type equations. A cumulative probability graph was used to determine an approximate reduced model. The split-plot error structure was then introduced into the reduced model by applying generalized least square equations with variance components calculated using the restricted maximum likelihood approach. A model was developed to calculate the number of peaks observed with the chromatographic detector at 210 nm. A 20-term model contained essentially all the statistical information of the initial model and had a root mean square calibration error of 1.38. The model was used to predict the number of peaks eluted in chromatograms obtained from extraction solutions that correspond to axial points of the simplex centroid design. The significant model coefficients are interpreted in terms of interacting linear, quadratic and cubic effects of the mobile phase and extraction solution components.

Simplified cryopreservation of porcine cloned blastocysts.

Recently, a non-invasive delipation (lipid removal) method combined with ultrarapid vitrification has been used successfully for in vitro produced (IVP) porcine embryos. In the present study, this method was combined with parthenogenesis and a recent form of somatic cell nuclear transfer (SCNT) - handmade cloning (HMC) - to establish a simplified and efficient cryopreservation system for porcine cloned embryos. In Experiment 1, zonae pellucidae of oocytes were partially digested with pronase, followed by centrifugation to polarize lipid particles. Ninety percent (173/192) oocytes were successfully delipated in this way. Parthenogenetic activation (PA) after complete removal of zona resulted in similar blastocyst rates in delipated vs. control oocytes (28+/-7% vs. 28+/-5%, respectively). Subsequent vitrification of produced blastocysts with the Cryotop technique resulted in higher survival rates in the delipated group compared to the control group (85+/-6% vs. 32+/-7%, respectively; P<0.01). In Experiment 2, delipated oocytes were used for HMC with normal oocytes as control. Partial zona digestion was further applied before enucleation both in delipated and control groups, to bisect oocyte successfully. Although the blastocyst rate of reconstructed embryos was similar between groups derived from delipated vs. control oocytes (21+/-6% and 23+/-6%, respectively), after vitrification higher survival rates were achieved in the delipated groups than in controls (79+/-6% vs. 32+/-8%, respectively). Our results prove that porcine embryos produced from delipated oocytes by PA or HMC can be cryopreserved effectively by ultrarapid vitrification. Further experiments are required to assess the in vivo developmental competence of the cloned-vitrified embryos.

In vitro activity evaluation of Parkia pendula seed lectin against human cytomegalovirus and herpes virus 6.

Human cytomegalovirus (HCMV) in vitro infectivity was inhibited by Parkia pendula seed lectin (PpeL) in contrast to human herpes virus 6 (HHV-6) which was not affected. The antiviral activity was detected for HCMV in human embryo lung (HEL) cells using a microtechnique in culture plates. The assay showed a reduction of cellular infectivity from approximately 95%, at a concentration of 150microg/mL with minimal cytotoxicity (25%). Also, a reduction of 75% was observed in HEL cells at a concentration of 75microg/mL without toxic effect. The reduction on infectivity was observed even after virus pre-adsorption to cells suggesting that this action should occur after virus penetration, in the intracellular replication phase. MT4 lymphocytes and cord blood mononuclear cells (CBMC) were used to evaluate the lectin effect on HHV-6 following the same technique. Lectin concentrations with few or no toxic effects on lymphocytes did not show inhibitory action of HHV-6 cytopathic effect. The results obtained with PpeL demonstrate that it may have an impact in the design of pharmacological strategies to infection of cytomegalovirus.

Exploratory analysis of simultaneous degradation of anthocyanins in the calyces of flowers of the Hibiscus sabdariffa species by PARAFAC model.

A three-way resolution method based on PARAFAC model was applied for the UV-Vis spectra to study the simultaneous degradation of anthocyanins extracted from fresh calyces of flowers of the Hibiscus sabdariffa. This methodology was used to resolve a chemical system, for which there was no a priori information about the composition or the pure spectra, a so-called black system. In the pH range from 1 to 13, seven species were identified: flavylium cation, carbinol, quinoidal base, E- and Z-chalcones and E- and Z-ionized chalcones. The concentration changes were determined as functions of pH at different wavelengths. The pK values for the acidity constants as well as tautomeric constant were estimated as 2.70, 3.54 and 0.14, respectively. The spectral profiles recovered by the chemometric methods are in excellent agreement with bands of experimental spectra reported in the literature for the species measured at specific pH values.

Simultaneous spectrophotometric determination of Hg2+, Pb2+, and Cu2+ by Zn(dmit) using the multivariate calibration method.

This work investigates the applicability of the anionic complex [NBu4]2[Zn(1,3-dithiol-2-tione-4,5-dithiolate)2] on the simultaneous spectrophotometric determination of transition metals. Principal Component Regression was used in the data analysis. This method was used to determine Hg2+, Cu2+ and Pb2+ in water solution, at t = 0, 20 and 60 min of reaction. The root mean square errors (rms) for Hg2+, Cu2+, Pb2+ were 1.92 x 10(-7), 1.89 x 10(-7) and 9.35 x 10(-8) for t = 0 min, 1.79 x 10(-7), 2.1 x 10(-7) and 2.22 x 10(-7) for t = 20 min and 1.22 x 10(-7), 1.31 x 10(-7) and 1.10 x 10(-7) for t = 60 min, respectively. Results showed the viability of the method in determining concentrations of metallic ions in aqueous medium with high spectral overlap. It is an efficient alternative method for exploring the kinetic behavior of multicomponents systems of interest in physicochemical research.

Three-way chemometric method study and UV-Vis absorbance for the study of simultaneous degradation of anthocyanins in flowers of the Hibiscus rosa-sinensys species.

Ultraviolet-visible spectra of flower extracts of the Hibiscus rosa-sinensys L. var. regius maximus species have been measured between 240.02 and 747.97nm at pH values ranging from 1.1 to 13.0. Deconvolution of these spectra using the Parallel Factor Analysis (PARAFAC) model permitted the study of anthocyanin systems without isolation and purification of the individual species. Seven species were identified: flavilium cation, carbinol, quinoidal base, and E and Z-chacone and their ionized forms. The concentration changes of flavilium cation, quinoidal basen, and E and Z ionized chalcones were determined as function of pH at the different wavelengths. The flavilium cation, quinoidal base, and ionized E-clacone are involved in tho stage kinetic processes, a fast one followed by a slower one. Ionized Z-chalcone obeys a simple first-order processes. The spectral profiles recovered by PARAFAC model are in excellent agreement with bands of experimental spectra reported in the literature for the individual species measured at specific pH values. These results complement those obtained using chemical and simple mathematical techniques and demonstrate how chemometric methods can resolve problems for complex systems.

Comparison of metal analysis in sediments using EDXRF and ICP-OES with the HCl and Tessie extraction methods.

The work presents an investigation on metal availability in sediments during 13 months using the dispersive-energy X-ray fluorescence (EDXRF) and atomic emission spectrometry with induced argon plasma (ICP-OES) techniques and single extraction (0.1moll(-1) HCl) and Tessie's sequential speciation methods. The EDXRF technique could yield essentially the same profile as ICP-OES for the seasonal variation of metals in sediments, but in a more practical way. The sequential extraction procedure (SEP) was more efficient in metal dissolution than single extraction. The Pb, Ni, Al, Cr, and Fe elements were less efficiently extracted with single extraction in relation to sequential extraction. For Co both methodologies were equivalent, but for Cu and Mn the extraction was higher with single extraction. Single extraction does not mobilize Pb, Ni, Al, Cr, and Fe adsorbed on oxides and bound to organic matter. However for Cu and Mn, not only extracted these metals from the four fractions, but it also dissolved part of the fifth fraction (residual). Principal Component Analysis discriminated seasonal variations in the content of several metals, mainly Fe, Co, Ni, and Zn. The mobility of metallic ions in the sediments is conditioned to the seasonal flow of organic and inorganic material coming from the river or by the erosion of adjacent soils.