Evaluation of isoflavone aglycon and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector

An ultrafast HPLC/UV-vis DAD method working at 254 nm was applied for the determination of isoflavone aglycons and glycosides (genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin, and biochanin A) in roots, stems, leaves, and soy pods of soy plants and in soybeans of five varieties (Korada, Quito, Rita, OAC Erin, and OAC Vison). An Atlantis dC18 ultrafast RP chromatographic column (20 mm x 2.1 mm, 3 microm particle size) was applied for separation of the isoflavone aglycons and glycosides. A flow rate of the mobile phase (0.1% (v/v) acetic acid, pH 3.75-solvent A and methanol-solvent B) was 0.35 mL min(-1), and the column temperature was 36 degrees C. A linear gradient profile from 13 up to 22% B (v/v) from zero to 2.5 min, up to 30% B to 3.21 min, up to 35% B to 4 min, up to 40% B to 4.5 min, up to 50% B to 5.14 min, and followed by negative gradient up to 13% B to 7.71 min was used. The absolute limits of detection per sample injection (5 microL) were the highest for biochanin A (166.2 fmol) and the lowest for genistin (17.0 fmol), respectively. An accelerated solvent extraction (ASE) in combination with sonication was applied for isolation of biologically active compounds. A solid-phase extraction procedure was used to purify the extracts in the case of analysis of soy plants parts. The recoveries of 96-106% were obtained for the different concentrations of the isoflavone aglycons and glycosides and the different matrixes (overall RSDs 2-9%). The highest isoflavone concentrations were found in roots (12.5 microg g(-1) dry weight), while the amounts were about 3-1100 microg g(-1) fresh weight in different varieties of soybeans.     

Chemometric studies of vinegars from different raw materials and processes of production

The phenolic composition, aroma compounds, and organic acid content of 83 vinegars have been determined. Multivariate analysis techniques have been used to classify these vinegar samples according to raw material (white wine, red wine, apple, honey, alcohol, balsamic, and malt) and production process (with and without aging in wood). Cluster analysis grouped the samples according to production process. Only apple and balsamic vinegars were separated from wine vinegars. Alcohol, honey, and malt vinegars were grouped with no aged wine vinegars. Linear discriminate analysis allowed a 88% differentiation according to raw material and 100% according to aging in wood. Besides, from the results obtained, a major role of the volatile compounds in the differentiation of the vinegar samples according to their aging period in wood can be seen.     

Olive fruit cell wall: degradation of pectic polysaccharides during ripening

Olive fruits at three stages of ripening (green, cherry, and black) have been studied. After cell wall isolation, the compositions of the cell wall and that of the phosphate-soluble polysaccharides were determined. In cell walls, decreases in arabinose, xylose, glucose, and uronic acid levels were observed, together with a slight increase in mannose on ripening. At the beginning of ripening, fragments of pectic polymers were the major constituents of the phosphate-soluble fraction, with the hemicellulosic ones increasing toward the end of the process. The molecular weight of the fragments solubilized was approximately 6 kDa. After cell wall fractionation, the pectic polysaccharides soluble in imidazole and sodium carbonate were also studied. In both fractions, between the green and cherry stages of ripening, a significant loss of homogalacturonans took place. Between the cherry and black stages of ripening, rhamnogalacturonan side chains were also released in addition to homogalacturonans. In any of the pectic fractions, changes in apparent molecular weight were quantified.     

Effect of mycorrhizal inoculation and soil restoration on the growth of Pinus halepensis seedlings in a semiarid soil

An experiment was carried out to evaluate the growth of mycorrhizal Pinus halepensis seedlings planted in a semiarid soil amended with urban refuse in southeast Spain. Three fungal species were used: Pisolithus tinctorius, Rhizopogon roseolus, and Suillus collinitus. After 8 months, inoculated seedlings grown under controlled conditions did not differ significantly from controls with regard to plant height and nutrient assimilation. Other features such as root development and stem dry weight showed that the plants grew better in the absence of mycorrhizal inoculation. The mycorrhizal seedlings and the controls were planted in three experimental plots treated with urban refuse (0, 6, and 12 Kg m^-2). After 1 year of growth under field conditions the results showed that the type of fungus inoculated significantly influenced P. halepensis development. This effect varied with the dose of urban refuse. Plant growth was encouraged by the application of refuse but only at the lowest dose. Under these conditions P. tinctorius was the most effective fungus and R. roseolus yielded poorer plant development. The highest application of urban refuse led to notably worse results and a significant decrease in seedling growth compared to controls. In the control plot (without refuse) S. collinitus was the most effective fungus in plant growth improvement. The smallest application of urban refuse had a positive effect on the assimilation of N, P, and K in seedlings inoculated with P. tinctorius and S. collinitus.     

Usefulness of the frequency data of the fourier transform infrared spectra to evaluate the degree of oxidation of edible oils

The oxidation process of 13 edible oil samples with different proportions of oleic, linoleic, and linolenic acyl groups has been studied using Fourier transform infrared spectroscopy. The oxidation experiments were carried out by heating the samples in a convection oven at 70 degrees C. Duplicate spectra were recorded from a film of pure oil between two disks of KBr for each sample every day during the course of the oxidation, and frequency data of each band of the spectrum were collected automatically. Changes in the values of the frequency of most of the bands of the spectra were observed. The shiftings of the frequency value of specific bands allowed one to distinguish between the different stages of the oxidation process and to establish the oxidation degree of each oil sample. This methodology could be useful to evaluate the oxidative stability of edible oils in a simple, fast, and accurate way.     

Spatial patterns of diversity and genetic erosion of traditional cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis> Crantz) in the Peruvian Amazon: An evaluation of socio-economic and environmental indicators

This study evaluates quantitatively the suitability of the use of site-specific socio-economic and environmental data as indicators to rapidly assess patterns of diversity and genetic erosion risk in cassava. Socio-economic data as well as farmers’ estimation of genetic erosion were collected in the study area, the Ucayali region of the Peruvian Amazon, through interviews with 285 cassava farmers in 50 communities, while diversity was assessed based on agromorphological characterization of 295 cassava accessions. Using multivariate regression analyses, 50 and 45% of the variation in respectively diversity and genetic erosion estimation could be explained by a selected set of socio-economic and environmental indicators. In both regression models four out of the total of 38 variables proved to contribute significantly (at p < 0.10 level). Additionally, the study revealed that farmers are a good direct source of information on the diversity present at community level, which can contribute to the development of methodologies to assess diversity more rapidly. The results of this study are valuable for the development of models to rapidly assess diversity dynamics in large areas.     

One-sided tests of bioequivalencewith nonnormal distributions and unequal variances

A test for bioequivalence on a variable of interest is sometimes used to determine whether a site that may have been adversely affected by its past use is sufficiently similar to a control site for the two to be considered the same for all practical purposes. For example, such a test might be used to compare the mean percentage cover of vegetation in an area previously used for mining and subsequently reclaimed, with the mean of the same variable at a neighboring area that has never been mined. A reason for adopting this approach rather than a standard test of significance is that it reverses the burden of proof. A significant result for a bioequivalence test gives evidence for two sites being similar, while a nonsignificant result leads to the conclusion that there is not enough evidence to consider the sites to be similar. This article discusses how to carry out one-sided tests for bioequivalence when the data are from nonnormal distributions, with variances that may be unequal. Examples that are used to illustrate the proposed methods involve the measurement of arsenic levels at an old ammunition testing site, and the vegetation cover at a reclaimed mining site.     

Establishment of Retama sphaerocarpa L. seedlings on a degraded semiarid soil as influenced by mycorrhizal inoculation and sewage‐sludge amendment

A field experiment was carried out to evaluate the effectiveness of mycorrhizal inoculation with three arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge), and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and the addition of composted sewage sludge (SS) with respect to the establishment of Retama sphaerocarpa L. seedlings, in a semiarid Mediterranean area. Associated changes in soil chemical (nutrient content and labile carbon fractions), biochemical (enzyme activities), and physical (aggregate stability) parameters were observed. Six months after planting, both the addition of composted SS and the mycorrhizal‐inoculation treatments had increased total N content, available‐P content, and aggregate stability of the soil. Values of water‐soluble C and water‐soluble carbohydrates were increased only in the mycorrhizal‐inoculation treatments. Rhizosphere soil from the mycorrhizal‐inoculation treatments had significantly higher enzyme activities (dehydrogenase, protease‐BAA, acid phosphatase, and β‐glucosidase) than the control soil. In the short‐term, mycorrhizal inoculation with AM fungi was the most effective treatment for enhancement of shoot biomass, particularly with G. mosseae (about 146% higher with respect to control plants). The addition of the composted SS alone was sufficient to restore soil structural stability but was not effective with respect to improving the performance of R. sphaerocarpa plants.     

Evolution of humic substances from unripe compost during incubation with lignolytic or cellulolytic microorganisms and effects on the lettuce growth promotion mediated byAzotobacter chroococcum

Some recalcitrant organic wastes, which contain a large proportion of lignin or cellulose, are not changed much by composting, and thus the effectiveness of the compost as fertilizer is usua.lly low. In this study, incubation of unripe compost with ligno-cellulolytic microorganisms —Trichoderma viride orBacillus spp. — was investigated to increase the degree of humification of the organic matter present, and improve ils quality as a soil amendment. High-performance liquid chromatography (HPLC) analyses together with humification indices and electrofocusing patterns were used to monitor the evolution of the humic substances during the incubation process. Plant growth effects exerted by Azotobacter chroococcum on lettuce plants growing on the previously incubated compost were affected by the length of incubation and by changes in the composition of humic substances. Higher organic matter content and better humification seem to be important factors for predictingA. chroococcum hebaviour in the rhizosphere.