Further studies on the analysis of DSP toxin profiles in galician mussels

Further studies on mussel samples from Galicia, Spain, have revealed the presence of okadaic acid (OA), dinophysistoxin 2 (DTX2), and the fatty acid acyl esters of both of these toxins as the "DTX3" complex. Measurements were performed with an improved in situ method for the formation of 9-anthryldiazomethane (ADAM) derivatives followed by liquid chromatography with fluorescence detection. Base hydrolysis of DTX3 toxins gave free OA and DTX2, which were determined following ADAM derivatization. Results were confirmed by liquid chromatography/mass spectrometry analyses, and in most of the samples, free DTX2 was the most abundant toxin. However, the OA/DTX2 ratio in the DTX3 conjugated form was different, with OA being the most abundant in all cases. This difference could be due to different rates of metabolism of OA and DTX2 to the acyl esters or due to contamination of the shellfish by the two toxins at different points in time, resulting in less acyl ester formation for one toxin versus the other. The second possibility would be reasonable if two different source organisms were producing the toxins.     

High-resolution NMR and diffusion-ordered spectroscopy of port wine

The use of high-resolution NMR and high-resolution diffusion-ordered spectroscopy (DOSY) for the characterization of selected Port wine samples of different ages with the aim of identifying changes in composition is described. Conventional 1D and 2D NMR methods enabled the identification of about 35 compounds, including minor components such as some medium-chain alcohols, amino acids, and organic acids. High-resolution (HR) DOSY extended sample characterization, increasing the number of compounds identified and NMR assignments made, by providing information on the relative molecular sizes of the metabolites present. Port wines of different ages were found to differ mainly in their content of (a) organic acids and some amino acids, (b) an unidentified possible disaccharide, and (c) large aromatic species. The relative amount of these last high Mw aromatics is seen to decrease significantly in the oldest wine, as expected from the known formation and precipitation of anthocyanin-based polymers during red wine aging.     

Source of polynuclear aromatic hydrocarbons found in sediment in a region of expanding sugarcane cultivation of São Paulo State,Brazil

Purpose

Polycyclic aromatic hydrocarbons (PAHs) are unintentional products that are classified as persistent toxic substances. The goal of the present study was to generate data on the presence of 15 priority PAHs that are found in surface sediment and core sediment in the region of the Turvo/Grande watershed, São Paulo State, Brazil, which is an area of expanding sugarcane cultivation, and to correlate these data with the sources of these PAHs and the guiding values for sediment quality analysis.

Materials and methods

Surface sediments and sediment cores were sampled during the rainy and dry seasons in February and July 2010. The extraction of PAHs from sediments was performed using a Soxhlet extractor, and then the extract was cleaned according to the methods of the US EPA 3630C (US EPA 1996) using a silica gel column. Quantification was performed using high performance liquid chromatography with fluorescence detection.

Results and discussion

The concentrations of all 15 PAHs decreased as the depth of the sediment core increased. Overall, the concentrations decreased along the sediment core; however, the RTURARG (region predominantly used for planting sugarcane and livestock) during the rainy season and the CAPRP sampling site (located at part of the Preto River dam) during the dry season showed increased concentrations in the first few sediment fractions, and then the concentrations decreased. Higher concentrations were observed in urban locations, and the concentration of naphthalene was higher than the probable effect level (PEL) determined by the Canadian environmental agency. The obtained diagnostic ratios indicate that the sediment from areas with an abundance of sugarcane was a pyrolytic source of PAHs, which indicates a contribution from burning straw to the PAH concentrations in those areas.

Conclusions

For all sampling sites and all PAHs, we found a decreasing trend in PAH concentrations with increasing sediment core depth, and the locations, such as CAPRP, that experienced a higher level of human activity had the highest total concentrations of PAHs. These locations were the only areas in which the PAH naphthalene was found in higher concentrations than the PEL. The diagnostic ratios reveal that regions with sugarcane plantations had predominantly pyrolytic sources of PAHs, indicating the contribution of PAHs from sugarcane straw burning.

     

Quantitation of short-chain glycerol-bound compounds in thermoxidized and used frying oils. A monitoring study during thermoxidation of olive and sunflower oils

Major short-chain glycerol-bound compounds were investigated in olive oil (OO) and conventional sunflower oil (SO) during thermoxidation at 180 degrees C for 5, 10, and 15 h. These compounds included methyl heptanoate (C7:0), methyl octanoate (C8:0), methyl 8-oxo-octanoate (8-oxo-C8:0), methyl 9-oxononanoate (9-oxo-C9:0), dimethyl octanodiate (C8:0 diester), and dimethyl nonanodiate (C9:0 diester), which were analyzed by GC after derivatization of triacylglycerols to fatty acid methyl esters. An acceptable linear correlation (r = 0.967) was found between the total content of these compounds and the total content of polar compounds, suggesting that quantitation of the major short-chain glycerol-bound compounds provides a good indication of the total alteration level of oils heated at frying temperature. Samples with levels of polar compounds around 25% on oil showed total contents within 2-3 mg/g of oil. To determine the content of these compounds in used frying oils, 10 samples from restaurants and fried-food outlets in Spain were analyzed. Results showed total levels between 2.13 and 7.56 mg/g of oil in samples with contents of polar compounds ranging from 18.8 to 55.5% on oil. Samples with levels of polar compounds of approximately 25% showed total contents of the short-chain compounds similar to those found in the thermoxidized oils, that is, within 2-3 mg/g of oil.     

Antioxidant activity of ethanolic extracts from several asparagus cultivars

Three different methods (antiradical activity, inhibition of primary oxidation, and ferric reducing power) have been used to evaluate the antioxidant activity of eight different asparagus cultivars and byproducts: white and green asparagus from Alcalá del Río (Guadalquivir Valley, Seville) and American hybrids, native spears, and their byproducts from Huétor-Tájar (Vega de Granada). The correlation between antioxidant activity and total phenol content was studied. Six standards were also tested to validate the modified methods for antioxidant activity determination. Results obtained for antiradical capacity and reducing power were very similar, and a high correlation with phenols was found (R > or = 0.9 for both tests). Sample origin was an important factor, spears from Huétor-Tájar having higher values (ARC between 7 and 10 and P(R) of 0.25-0.33) than those from Alcalá del Río (ARC 0.6-2 and P(R) of 0.05-0.07). Significant differences were found between spears with the same origin, suggesting that genetics are another factor to take into account. Asparagus inhibits lipid primary oxidation, but no correlation between the inhibition percentage and phenols was observed. Asparagus origin was the only factor that led to significant differences: samples from Huétor-Tájar had higher values (POIC between 18 and 32) than those from Alcalá del Río (POIC of 5-9). Byproducts from the canning industry at Huétor-Tájar were also assayed for antioxidant activity; the results obtained suggested that byproducts could be considered as an excellent source of natural antioxidants.     

Pectins as possible source of the copper involved in the green staining alteration of cv. Gordal table olives

The pectic and pigment compositions and Ca and Cu contents of the alcohol-insoluble solid (AIS) residues were determined in cv. Gordal olives treated with NaOH solution and kept at different constant pH values (3.5-6.5). The same controls were made in table olives presenting green staining alteration. The ratio between the various pectin fractions of the more acid pH experiment samples remained similar in fruits not showing green staining. In altered fruits, the protopectin fraction was lower, and the calcium pectate or EDTA soluble pectins were higher. Regarding the presence of Ca and Cu in the AIS, it was observed that, whereas Ca levels fell at the most acid pH values, those of Cu increased. The concentration of Ca was higher in the AIS of altered olives than in nonaltered ones. The same trend was seen for the zone with or without green staining of an altered fruit. In the case of Cu, the relationship was the opposite: a decrease in the levels of AIS Cu in fruits and zones of fruits with green staining. This result was correlated with the highest concentration of Cu-chlorophyll complexes found in such samples and suggested that pectins might act as a reservoir of Cu involved in the alteration.     

Purification and properties of a neutral peroxidase isozyme from turnip (Brassica napus L. Var. Purple Top White Globe) roots

A neutral peroxidase isozyme (pI 7.2) from turnip roots (TNP) was purified to homogeneity and partially characterized. TNP is a monomeric glycoprotein with 9.1% carbohydrate content and a molecular weight of 36 kDa. Optimum pH values for activity using 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and guaiacol as H donors were 4.5 and 5.5, whereas the K(m) values were 0.7 and 3.7 mM, respectively. The ABTS K(m) was approximately 7 times higher than that reported for basic commercial horseradish peroxidase (HRP-C). TNP retained approximately 70% activity after 11 min of heating at 65 degrees C, whereas the activation energy for inactivation (132 kJ/mol) was higher than or comparable to that of other peroxidases. The low ABTS K(m) and high specific activity (1930 units/mg) gave a high catalytic efficiency (500 M(-1) s(-1)). These properties make TNP an enzyme with a high potential as an alternative to HRP in various applications.     

Modified method for the analysis of free fatty acids in fish

The objective of this study was the development of a method for the quantification of free fatty acids (FFA) using less aggressive reactants against the handler and the environment than those used in the classic method of Lowry and Tinsley. The modified procedure is a variation of the Lowry and Tinsley method employing cyclohexane in place of benzene. The use of benzene is prohibited in certain work processes and laboratories, and the competent authority in each country is actively promoting research into harmless or less harmful products that could replace benzene. A comparison with the traditional AOCS titration method for oil analysis was performed. FFA content in mackerel frozen at -10 degrees C was measured according to the three methods over a 12 month period. The results showed similar values, and good correlations were obtained.     

Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds

Dietary ellagic acid and related polyphenols are metabolized in humans to dibenzopyran-6-one derivatives, and the microbial origin of these metabolites has been suggested. However, this has not been demonstrated so far. Fecal samples donated by six volunteers were incubated under anaerobic conditions, and aliquots were used to evaluate the fecal metabolism of ellagic acid, the ellagitannin punicalagin, and an ellagitannin rich extract from walnuts. The isoflavone daidzein was also incubated with the same fecal samples to follow the production of the microbial metabolites previously reported (dihydrogenistein, O-demethylangolensin, and equol) as a positive control of the system and to evaluate similarities between isoflavone and ellagic acid fecal flora metabolism. After fermentation the metabolite "urolithin A" (3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one) was produced from ellagic acid, punicalagin, and the ellagitannin extract in all the fecal cultures from different volunteers, but with very different production rates and concentrations. This large variability in the concentration of metabolite and kinetics of metabolite production is consistent with the large variability found in the excretion of these metabolites in urine in vivo after human consumption of ellagitannins, and with differences in the composition of the fecal microflora. No correlation between isoflavone and ellagic acid metabolism by fecal microflora was observed. The present study confirms the microbial origin of the recently reported in vivo generated hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives in humans and is a further step in the study of the bioavailability and metabolism of ellagic acid and ellagitannins.     

Adsorption of pesticides from water by functionalized organobentonites

Replacement of natural inorganic cations of clay minerals with organic cations has been proposed as a strategy to improve the adsorptive capacity of clay minerals for organic compounds, including pesticides. The organic cations most commonly used for this purpose have been quaternary ammonium ions containing alkyl or aryl chains without specific functional groups. In this work, we evaluated the ability of two bentonites (SWy-2 and SAz-1) exchanged with four natural organic cations containing diverse functional groups (L-carnitine, L-cysteine ethyl ester, L-cystine dimethyl ester, and thiamine) as adsorbents of pesticides varying in their chemical structures (simazine, hexazinone, triadimefon, alachlor, carbaryl, and imazethapyr). For comparison purposes, the adsorptive properties of two "classical" organobentonites, hexadecyltrimethylammonium- and phenyltrimethylammonium-exchanged bentonites, were also determined. Most organobentonites displayed higher affinity for the pesticides than the untreated bentonites, but the improvement in adsorption capacity varied depending on the characteristics of the pesticide and the interlayer organic cation. Triadimefon, carbaryl, and imazethapyr displayed the highest affinity for carnitine (K(f) = 229-2377)-, thiamine (K(f) = 83-354)-, and cystine (K(f) = 96-100)-treated bentonites, respectively, whereas alachlor was adsorbed similarly by all organobentonites. In general, pesticide adsorption-desorption hysteresis was greater for adsorbents with the highest adsorption capacities. The results demonstrate that selective modification of smectitic clay minerals with natural organic cations containing appropriate functional groups can be a useful strategy to improve their performance for the removal of specific pesticides from the environment.