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.     

Characterization and quantitation of phenolic compounds in new apricot (Prunus armeniaca L.) varieties

Thirty-seven apricot varieties, including four new releases (Rojo Pasión, Murciana, Selene, and Dorada) obtained from different crosses between apricot varieties and three traditional Spanish cultivars (Currot, Mauricio, and Búlida), were separated according to flesh color into four groups: white, yellow, light orange, and orange (mean hue angles in flesh were 88.1, 85.0, 77.6, and 72.4, respectively). Four phenolic compound groups, procyanidins, hydroxycinnamic acid derivatives, flavonols, and anthocyanins, were identified by HPLC-MS/MS and individually quantified using HPLC-DAD. Chlorogenic and neochlorogenic acids, procyanidins B1, B2, and B4, and some procyanidin trimers, quercetin 3-rutinoside, kaempferol 3-rhamnosyl-hexoside and quercetin 3-acetyl-hexoside, cyanidin 3-rutinoside, and 3-glucoside, were detected and quantified in the skin and flesh of the different cultivars. The total phenolics content, quantified as the addition of the individual compounds quantified by HPLC, ranged between 32.6 and 160.0 mg 100 g(-1) of edible tissue. No correlation between the flesh color and the phenolic content of the different cultivars was observed.     

Application of hexanal,E-2-hexenal,and hexyl acetate to improve the safety of fresh-sliced apples

The aims of this work were to evaluate the effects of different concentrations of hexanal, (E)-2-hexenal, hexyl acetate, and their mixtures on the fate of pathogenic species such as Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes inoculated in model systems as well as the antimicrobial activity against the target species of the chosen molecules when added to the packaging atmosphere of inoculated fresh-sliced apples. The result obtained in this work pointed out the potential use of compounds such as hexanal, (E)-2-hexenal, and hexyl acetate for both the extension of shelf life and an improvement of hygienic safety of "minimally processed foods". In fact, hexanal, (E)-2-hexenal, and hexyl acetate had a significant inhibitory effect against pathogen microorganisms frequently isolated from raw materials (E. coli, S. enteritidis, and L. monocytogenes) when inoculated in both model and real systems. In this last condition, these compounds, at the levels used (150, 150, and 20 ppm for hexanal, hexyl acetate, and (E)-2-hexenal, respectively), displayed a bactericide effect on L. monocytogenes and they exhibited significant extensions of lag phase of E. coli and S. enteritidis inoculated at levels of 10(4)-10(5) CFU/g.     

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.     

Genetic variability evaluation in a Moroccan collection of barley, Hordeum vulgare L., by means of storage proteins and RAPDs

The genetic variation existing in a set of barley (Hordeum vulgare L.) landrace samples recently collected in Morocco was estimated. Two kinds of genetic markers, seed storage proteins (hordeins) and random amplified polymorphic DNA (RAPD), were used. Only six out of 31 landraces were subjected to RAPD analysis. Both kinds of markers, RAPD and storage proteins, yielded similar results, showing that the level of variation observed in Moroccan barley was high: all landraces showed variability; 808 different storage protein patterns (multilocus associations) were observed among 1897 individuals (2.32 seeds per association, on average) with an average of 43 multilocus associations per accession. In general, genetic variation within accessions was higher than between accessions. The 100 polymorphic RAPD bands generated by 21 effective primers were able to generate enough patterns to differentiate between uniform cultivars and even between individuals in variable accessions. One of the aims of this work was to compare the effectiveness of RAPD versus storage protein techniques in assessing the variability of genetic resource collections. On average hordeins were more polymorphic than RAPDs: they showed more alternatives per band on gels and a higher percentage of polymorphic bands, although RAPDs supply a higher number of bands. Although RAPD is an easy and standard technique, storage protein analysis is technically easier, cheaper and needs less sophisticated equipment. Thus, when resources are a limiting factor and considering the cost of consumables and work time, seed storage proteins must be the technique of choice for a first estimation of genetic variation in plant genetic resource collections.     

A new yam bean (Pachyrhizus spp.) interspecific hybrid

The yam bean (Pachyrhizus spp.) is a legume tuber / root crop locally grown in Central America, South America and Asia. The tuber is usually consumed raw for its refreshing taste and high moisture content. A recently found P. tuberosus type from a small ecogeographic region in the tropical lowlands of Peru has a high tuber dry matter content and this so-called Chuin-Type is consumed like manioc. This study was conducted to determine the possibility to use the Chuin-Type as a source to incorporate high dry matter into the remaining yam bean gene pool. Three P. tuberosus (Chuin) accessions were crossed with 15 P. ahipa accessions from the Andean highlands resulting in six successful interspecific hybridisations. Successful crosses were confirmed by morphological and agronomical traits as well as multivariate statistics. All hybrid plants were fertile and vigorous. Owing to the fertility and vigour of interspecific hybrids it is assumed that a hybrid between P. tuberosus and P. ahipa might already evolved in the area of origin and that interspecific hybridisation is appropriate to improve the tuber dry matter content in the yam bean which might give the crop new forms of food and processing use.     

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.     

Chlorogenic acids as a potential criterion in coffee genotype selections

A systematic study by HPLC was conducted to determine the content of chlorogenic acids in green coffee beans of the Variedad Colombia (Coffea arabica Caturra var. x Híbrido de Timor) and in other genotypes of interest of C. arabica, Coffea canephora, Híbrido de Timor, and the F1 offspring derived from the crossing Caturra x Híbrido de Timor. Comparisons were made of the total content of these acids, their quantitative differences, and the presence or absence of some of them, using parametric statistical techniques and multivariate analysis. Total content differences were found between C. canephora accessions, between Típica and Caturra varieties of C. arabica, and between Híbrido de Timor accessions. The chromatographic profile comparisons with principal component analysis separated in the first component C. canephora accessions from the rest of the genotypes, whereas the second component separated C. canephora accessions.     

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.     

Determination of Organic Mercury in Biota, Plants and Contaminated Sediments Using a Thermal Atomic Absorption Spectrometry Technique

A simple, rapid procedure for the determination of organic mercury in sediments, plants and fish tissues has been developed and validated. Extraction and separation of organic mercury compounds from the sample matrix was achieved by an established procedure based on an acid leaching of the sample (H₂SO₄/KBr/CuSO₄), followed by extraction of the organic mercury halide with toluene and back-extraction with an aqueous solution of thiosulphate. Detection and quantification of mercury, in the liquid extracts, was made by atomic absorption spectrometry (AAS), following thermal decomposition of the sample. The method was evaluated using Certified Reference Material (CRM) BCR 463 (tuna fish), BCR 580 (estuarine sediment), IAEA-140TM (sea plant homogenate) and NRCC TORT-2 (lobster hepathopancreas). The recovery factors for organic mercury in all tested CRM were between 81–107%. The precision of the method has relative standard deviations of less than 10% for sediments and fish tissues and of less than 16% for plant material. The method was successfully applied to natural samples of sediments, plants, macroalgae and fish tissues collected from an estuarine ecosystem and could, therefore, be used for routine analyses.