Rapid liquid chromatography tandem mass spectrometry assay to quantify plasma (-)-epicatechin metabolites after ingestion of a standard portion of cocoa beverage in humans

A rapid liquid chromatography electrospray ionization tandem mass spectrometry with negative ion detection method was developed and validated to determine cocoa flavonoid metabolites in human plasma and urine after the intake of a standard portion of a cocoa beverage. A chromatographic run time of only 9 min provided clear separation of all metabolites and internal standards. Samples were analyzed in a product-ion scan of m/z 289, 369, and 465 to identify the metabolites and in multiple reaction monitoring acquisition mode to quantify (-)-epicatechin ((-)-Ec) (289/ 245), (-)-epicatechin-glucuronide ((-)-EcG) (465/289), and (-)-epicatechin-sulfate ((-)-EcS) (369/289). One (-)-Ec-G and three (-)-Ec-S were identified and confirmed in urine as the major metabolites, and one (-)-Ec-G was the only metabolite present in plasma volunteers (n = 5) at a mean concentration of 625.7 +/- 198.3 nmol/L at 2 h after consumption of a cocoa beverage containing 54.4 mg of (-)-Ec.     

Lead dietary intake in a Spanish population (Canary Islands)

For most people diet is the main route of exposure to trace metals, so information about dietary intake is also important to assess risks to human health for these elements. The purpose of this study was to determine the levels of Pb in the foods and drinks of highest consumption in the authors' our community to estimate daily intakes of Pb for each of the seven Canary Islands. Four hundred and twenty samples were analyzed using GFAAS. The total Pb intake of the Canarian population is 72.8 microg/day, 29.12% (for a person of 70 kg body weight) of the provisional tolerable weekly intake limit of 25 microg/kg fixed by the FAO/WHO. The island that presents the highest lead intake is La Gomera, followed by Lanzarote, Tenerife, and Gran Canaria islands. These four islands present a lead dietary intake over the mean Pb intake for the whole archipelago. The islands with lower Pb intakes are La Palma and Fuerteventura, with intakes under 70 microg/day. These results have also been compared with the values found for other national and international communities.     

Extracellular prolyl endoprotease from Aspergillus niger and its use in the debittering of protein hydrolysates

The observation that the bitterest peptides from casein hydrolysates contain several proline residues led us to hypothesize that a proline-specific protease would be instrumental in debittering such peptides. To identify the desired proline-specific activity, a microbiological screening was carried out in which the chromogenic peptide benzyloxycarbonyl-glycine-proline-p-nitroanilide (Z-Gly-Pro-pNA) was used as the substrate. An Aspergillus niger (A. niger) strain was identified that produces an extracellular proline-specific protease with an acidic pH optimum. On the basis of sequence similarities, we conclude that the A. niger-derived enzyme probably belongs to the S28 family of clan SC of serine proteases rather than the S9 family to which prolyl oligopeptidases belong. Incubating the overexpressed and purified enzyme with bitter casein hydrolysates showed a major debittering effect. Reversed phase HPLC analysis revealed that this debittering effect is accompanied by a significant reduction of the number of hydrophobic peptides present.     

Volatile components of Loureira, Dona Branca, and Treixadura wines.

White wines experimentally produced from the white grape varieties Loureira, Dona Branca, and Treixadura have been analyzed over four consecutive harvests. The contents of monoterpenes, volatile phenols, alcohols, sulfur components, acetates, fatty acids, and ethyl esters were determined by gas chromatography (FID) and gas chromatography-mass spectrometry. The mean values from four vintages confirmed that these wines have characteristic profiles. Loureira wines are characterized by a high content of free terpenes, 1-hexanol, ethyl acetate, and fatty acids. Dona Branca wines present the highest concentrations of methanol and sulfur constituents, and the lowest concentrations of higher alcohols, acetates, diethyl succinate, and ethyl esters. The levels of monoterpenes in Treixadura wines are very low, but they have the highest concentrations of volatile phenols, principally due to the vanillin, diethyl succinate, ethyl lactate, and ethyl esters. These results were confirmed by principal component and linear discriminant analysis, which show a clear differentiation among these wines as a function of the varietal origin.     

Determination of phytohormones of environmental impact by capillary zone electrophoresis

A test mixture of five phytohormones [naphthaleneacetic acid (NAA), naphthoxyacetic acid (NOA), indoleacetic acid (IAA), indolebutyric acid (IBA), and indolepropionic acid (IPA)] was investigated. These compounds were cleanly separated with good resolution by capillary zone electrophoresis with a UV diode array detector using 20 mM sodium phosphate buffer (pH 7.25). The lowest detection limit was obtained for IPA (0.45 mg L(-)(1) or 0.005 mg kg(-)(1)) and the highest for NAA (1.04 mg L(-)(1) or 0.014 mg kg(-)(1)). The method has been applied for tomato samples fortified with the five phytohormones using a liquid-liquid extraction procedure, obtaining recovery percentages ranging from 91 to 109.0%.     

Surface functional properties of native, acid-treated, and reduced soy glycinin. 2. Emulsifying properties

Emulsifying properties of native and chemically modified soy glycinins were studied. The influence of ionic strength, protein sample composition and concentration, and assay conditions on the flocculation-creaming process and coalescence resistance was analyzed. Differences in these emulsifying properties were exhibited by native glycinins, which have a variable content of 4S, 11S, and 15S forms. Structure and functionality of native glycinin were modified by means of combined treatments: mild acidic treatments without heating or with heating at variable time and with or without disulfide bonds reduction. Modified glycinins presented different degrees of deamidation, surface hydrophobicity, and molecular mass. A slight enhancement of emulsifying stability at moderated deamidation degrees was observed. In different protein samples, a positive relationship between the flocculation-creaming rate constant and equilibrium oil volume fraction of emulsions with surface hydrophobicity was detected. A remarkable difference was observed between reduced and nonreduced samples, mainly with respect to behavior at low or high ionic strength.     

An Easy Method to Measure Total Particulate Hg in Water Without Chemical Digestion

Particle transport often plays an important role in trace metal cycling in aquatic systems. In this study, we propose a simple method to estimate total particulate Hg in water using combustion as a treatment prior to analysis as opposed to conventional chemical digestion. We also present a field validation of this method in a stream. We sampled water from a stream 24 and 48 h after a heavy storm to obtain two sets of water samples with contrasting particulate Hg levels. Measurements of particulate and dissolved Hg using our new direct combustion method compared favorably with those of total Hg in unfiltered samples (112 ± 15 %). U.S. E.P.A. standard method 1631 gave similar dissolved Hg levels but different particulate Hg levels (calculated as the difference between total and dissolved Hg concentrations) than those obtained with our new method. We suggest that this discrepancy is explained by the propagation of errors resulting from the several calculation steps used in the standard method. Our new method gives reliable results, without labor intensive and costly chemical digestion and analysis of filters.     

Potential antioxidant capacity of sulfated polysaccharides from the edible marine brown seaweed Fucus vesiculosus

Fucus vesiculosus was sequentially extracted with water at 22 degrees C (fraction 1 (F1)) and 60 degrees C (F2), and with 0.1 M HCl (F3) and 2 M KOH (F4) at 37 degrees C. Soluble fractions (42.3% yield) were composed of neutral sugars (18.9-48 g/100 g), uronic acids (8.8-52.8 g/100 g), sulfate (2.4-11.5 g/100 g), small amounts of protein (< 1-6.1 g/100 g), and nondialyzable polyphenols (0.1-2.7 g/100 g). The main neutral sugars were fucose, glucose, galactose, and xylose. Infrared (IR) spectra of the fractions showed absorption bands at 820-850 and 1225-1250 cm(-1) for sulfate. F1, F2, and F4 also exhibited an absorption band at 1425 cm(-1), due to uronic acids, and their IR spectra resembled that of alginate. F3 had an IR spectrum similar to that of fucoidan with an average molecular weight of 1.6 x 10(6) Da, calculated by molecular exclusion high-performance liquid chromatography. The presence of fucose in this polysaccharide was confirmed by (1)H NMR spectroscopy. This fraction showed the highest potential to be antioxidant by the ferric reducing antioxidant power (FRAP) assay, followed by the alkali- and water-soluble fractions. Sulfated polysaccharides from edible seaweeds potentially could be used as natural antioxidants by the food industry.     

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.     

Possible role of plant phenolics in the production of trichothecenes by Fusarium graminearum strains on different fractions of maize kernels

Four trichothecene-producing strains of Fusarium graminearum were grown on three maize grain fractions, whole grain, degermed grain, and the germ, to determine the effect of natural substrates on mycotoxin production. Monitoring the ergosterol content after 25 days of incubation indicated that fungal growth on all grain fractions was comparable. Trichothecene (TCT) production was highest on degermed grain, less on whole grain, and very low or nondetectable on the germ; similar results were found with four different strains. It was concluded that inhibitor(s) of TCT biosynthesis were present in maize germ. The presence of phenolic compounds was investigated in the different fractions. The hydroxamate 4-acetylbenzoxazolin-2-one (4-ABOA), a known inhibitor of mycotoxin production, was found in the degermed and whole grain fractions but not in the germ. Therefore, the TCT inhibition observed on the maize germ fraction used in our study is clearly not linked to 4-ABOA. Other soluble phenolic compounds were found at a much higher concentration in the germ than in the two other fractions. The inhibition property of the soluble ester-bound extracts was tested in liquid culture. A possible role for these compounds is discussed.