Fatty Acids in Dry Beans (Phaseolus vulgaris L.): A Contribution to Their Analysis and the Characterization of a Diversity Panel.

Dry bean (Phaseolus vulgaris L.) is a crop of high nutritional interest widespread throughout the world. This research had two objectives. On the one hand, the development and validation of an analytical method to quantify fatty acids in dry beans based on the extraction and derivatization in a single step and later quantification by gas chromatography. On the other, its application to characterize the fatty acid content in a diversity panel consisting of 172 lines. The method was successfully validated in terms of accuracy, precision and robustness. Among the 14 fatty acids that constitute the fatty acid profile of dry bean, the most quantitatively important were linolenic acid, the major fatty acid in all cases, with an average value of 6.7 mg/g, followed by linoleic acid (3.9 mg/g), palmitic acid (2.9 mg/g) and oleic acid (1.5 mg/g). The concentrations of fatty acids in dry bean were influenced by the gene pool, with the Mesoamerican gene pool showing a higher content of palmitic, stearic, linoleic and linolenic acids and the Andean gene pool a higher level of cis-vaccenic acid. Also, the expression of fatty acid content showed high heritability. The information generated constitutes a robust database of interest in food technology, nutrition and breeding programs.

The Phenolic Composition, Antioxidant Activity and Microflora of Wild Elderberry in Asturias (Northern Spain): An Untapped Resource of Great Interest.

The objective of this study is the characterization of the phenolic profile and antioxidant activity of elderberries (Sambucus nigra L.) from a collection of 79 wild specimens in northern Spain to assess variations in the species at the local level and evaluate its interest as a source of biocompounds. Also, a first study was carried out on the microflora present in this fruit, providing information relevant to its commercial exploitation. Moreover, the phenolic composition, antioxidant capacity and microbial composition in overripe fruits were determined, seeking a better use for this currently wasted resource. A wide variability in levels of phenolics was detected. Elderberries showed high antioxidant activity related to a high cyanidin derivative content, making them of interest to industry. Microflorae were present in very variable concentration ranges, so their levels should be monitored in those applications that require strict control. Overripe fruits are of interest as a source of anthocyanidins, since their concentration and antioxidant capacity remain after the optimal ripening period, promoting sustainability and a better use of natural resources. The database generated is of particular interest for further breeding trials based on the phenolic profile and antioxidant activity of the samples.

Genome-wide association study for the extractable phenolic profile and coat color of common bean seeds (Phaseolus vulgaris L.).

BACKGROUND: A large variation in seed coat colors and seed phenolic metabolites is present in common bean (Phaseolus vulgaris L.). The study of the relationships between seed coat color phenotype and the phenolic profile is an important step in the elucidation of the gene network involved in the phenylpropanoid biosynthetic pathway. However, this relationship is still poorly understood in this species. RESULTS: A genome-wide association study (GWAS) was used to investigate the genomic regions associated with the synthesis of 10 flavonoids (5 anthocyanins and 5 flavonols) and with 10 seed coat color traits using a set of 308 common bean lines of the Spanish Diversity Panel (SDP) which have been genotyped with 11,763 SNP markers.. A total of 31 significant SNP-trait associations (QTNs) were identified, grouped in 20 chromosome regions: 6 for phenolic metabolites on chromosomes Pv01, Pv02, Pv04, Pv08, and Pv09, 13 for seed coat color on chromosomes Pv01, Pv02, Pv06, Pv07, and Pv10, and 1 including both types of traits located on chromosome Pv08. In all, 58 candidate genes underlying these regions have been proposed, 31 of them previously described in the phenylpropanoid pathway in common bean, and 27 of them newly proposed in this work based on the association study and their homology with Arabidopsis anthocyanin genes. CONCLUSIONS: Chromosome Pv08 was identified as the main chromosome involved in the phenylpropanoid pathway and in consequence in the common bean seed pigmentation, with three independent chromosome regions identified, Phe/C_Pv08(2.7) (expanding from 2.71 to 4.04 Mbp), C_Pv08(5.8) (5.89-6.59 Mbp), and Phe_Pv08(62.5) (62.58 to 63.28 Mbp). Candidate genes previously proposed by other authors for the color genes V and P were validated in this GWAS. Candidate genes have been tentatively proposed from this study for color genes B and Rk on Pv02, Asp on Pv07, and complex C on Pv08. These results help to clarify the complex network of genes involved in the genetic control of phenolic compounds and seed color in common bean and provide the opportunity for future validation studies.

Analysis of Cyanogenic Compounds Derived from Mandelonitrile by Ultrasound-Assisted Extraction and High-Performance Liquid Chromatography in Rosaceae and Sambucus Families.

An analytical method for extraction and quantitative determination of amygdalin, prunasin, and sambunigrin in plant material is described. The method is based on extraction with high-power ultrasound (UAE), with acidified water as solvent and quantification by HPLC-DAD. The best extraction conditions were: 80% sonication amplitude, 55 s extraction time, 70% duty cycle, 0.1 g sample mass, and 10 mL acidified water (0.1% perchloric acid). Once developed, the method was validated in terms of accuracy and precision. Good linearity was obtained, with correlation coefficients exceeding 0.999 and the quantification limits ranged from 2.2 µg/g (amygdalin) to 9.6 µg/g (sambunigrin). The accuracy (recovery study) ranged between 90 and 104% and the reproducibility of the method was always <2.3% (RSD). Special attention should be paid to the ratio sample/solvent in samples with potential ß-glucosidase activity to avoid degradation of the cyanogenic glycosides (CNGs). The proposed method was used to evaluate the content of CNGs in kernels of Prunus genera, apple seeds, apple pomace, and different plant materials of Sambucus nigra.

Variation of Morphological, Agronomic and Chemical Composition Traits of Local Hazelnuts Collected in Northern Spain.

Hazelnut is a traditional crop in northern Spain, where it grows wild as well as being cultivated. A field collection of 41 local and 17 non-local accessions, including 15 well-known cultivars, was established at SERIDA in Villaviciosa, Spain. Here, phenotypic variation was documented for phenological and morphological traits and chemical composition. A large degree of variation for most morphological and phenological traits, except nut maturity date, was revealed. Estimates of broad-sense heritability were high (>0.75) for most of the assessed characters, except for the first male bloom date (0.65), male and female flowering periods (0.40, 0.31), kernel weight (0.69), and kernel percentage (0.33). Local accessions produced smaller nuts and kernels than well-known cultivars but with higher kernel percentage. Limited overlapping between the male and female flowering periods (dychogamy) was observed, except for 'Forcinas 1', 'Forcinas 2', and 'Morell'. The local accessions generally exhibited significantly later male and female flowering compared with the reference cultivars. The local materials showed similar nutritional values to those reported previously for hazelnut. Moreover, the local accessions presented average values similar to the non-local accessions for total fat, ash and carbohydrate contents, as well as energy value, but their protein contents were lower. Their oils were rich in functional compounds, such as unsaturated fatty acids (average: 90.1%), tocopherols (514 mg/kg) and squalene (294.3 mg/kg). A hierarchical clustering on principal components analysis grouped the accessions and differentiated eight local accessions from the rest, including the landrace 'Casina'. This finding provides potential new cultivars, as well as sources of desirable traits, for European hazelnut breeding programs.

Phenolic Content and Antioxidant Activity in Seeds of Common Bean (Phaseolus vulgaris L.).

Dry bean (Phaseolus vulgaris L.) is one of the most important pulses consumed in the world. Total phenolic content, total flavonoid content, total monomeric anthocyanin content and antioxidant capacity were determined, using ferric reducing antioxidant power and free radical scavenging activity, in 255 lines grown under the same environmental conditions. For all parameters analysed, there was a wide range of variability, with differences always above one order of magnitude. Phenolic compounds in beans with coloured coats were found to be more efficient antioxidants than those with completely white coats, and samples with more strongly coloured coats (red, cream, black, pink and brown) showed the highest antioxidant capacities. Based on the strong correlation detected between the variables, total phenolic content can be considered an appropriate indicator of antioxidant activity. The results provide a robust database for selecting those lines of greater functional and nutritional interest in terms of cultivation for direct consumption, for inclusions in food formulations or for use in future breeding programs.

Aromatic, olfactometric and consumer description of sweet ciders obtained by cryo-extraction.

Nine ciders obtained by cryo-extraction were analysed for chemical, olfactometric and sensory characteristics. Three types of ice apple juices and three autochthonous yeast strains were evaluated. The quantitative volatile profile is mainly influenced by the apple juice. Regarding the olfactometric profiles of the ice ciders, 23 odorants not previously found in Spanish still ciders, and described as sweet, spicy, fruity and floral were observed. Among these, it is worth mentioning 5 high-boiling point compounds found in an olfactometric zone where heavy, phenolic odours predominate in still ciders. The sensory descriptions obtained by Check-All-That-Apply classified the ice ciders mainly by their sweet taste and smoothness, these attributes being the most influential in the assessment of the overall quality of the ciders. The most highly valued ciders were described as the sweetest, most fruity ones, in agreement with their having the highest values for the ratio between total sugars and total acidity.

Characterization of extractable phenolic profile of common bean seeds (Phaseolus vulgaris L.) in a Spanish diversity panel.

Phenolic compounds are important bioactive compounds in common bean (Phaseolus vulgaris L.). The aim of this work was the characterization of extractable phenolic profile (corresponding to 12 hydroxycinnamic acids and derivatives, 13 anthocyanins and 15 flavonols) in a bean diversity panel constituted by 220 lines, all grown under the same environmental conditions. Hydroxycinnamic derivatives were detected in all samples, while anthocyanins and flavonols were not detected in samples with completely white seed coats. In general, lines with black seeds showed higher contents of anthocyanins, followed by some red-seeded lines, while notable levels of flavonols were detected in market classes, including those with yellow, pink, and cream seed coats. However, a clear relationship between phenolic composition and seed phenotype could not be established, indicating the great influence of the genotype. This wide variability in the phenolic profiles analyzed is of particular interest for further breeding trials and the selection of varieties on the basis of this group of compounds.

Determination of volatile compounds in cider spirits by gas chromatography with direct injection.

Two analytical methods based on gas chromatography with direct injection are described for the quantitative analysis of volatile compounds (acetals, aldehydes, esters, alcohols, and volatile phenols) in cider brandies. Analytes were divided into major, 15, and minor volatile, 24, compounds depending on their usual concentration in samples. Parameters usually tested for method validation are evaluated. Correlation coefficients are calculated to estimate linearity, obtaining values higher than 0.999. Detection limits range between 0.325 mg/L (1-propanol) and 1.663 mg/L (methanol) among the major volatile compounds and between 0.086 mg/L (ethyl 2-methylbutyrate) and 0.332 mg/L (ethyl tetradecanoate) among the minor volatiles. Mean recoveries ranged between 109% (ethyl lactate) and 95% (1-butanol) for major volatiles and between 109% (1-octen-3-ol) and 94% (ethyl 2-methylbutyrate) for minor volatiles, thus confirming the accuracy of both methods. Reproducibility for major volatiles is < 5.4% (furfural) in all cases and < 9.6% (hexyl acetate) for minor volatiles. Moreover, the accuracy of the methods is evaluated by analyzing a certified whisky and five samples from interlaboratory assays, generally obtaining results in accordance with previous values.

Phenolic profile of Asturian (Spain) natural cider.

The polyphenolic composition of natural ciders from the Asturian community (Spain), during 2 consecutive years, was analyzed by RP-HPLC and the photodiode-array detection system, without previous extraction (direct injection). A total of 16 phenolic compounds (catechol, tyrosol, protocatechuic acid, hydrocaffeic acid, chlorogenic acid, hydrocoumaric acid, ferulic acid, (-)-epicatechin, (+)-catechin, procyanidins B2 and B5, phloretin-2'-xyloglucoside, phloridzin, hyperin, avicularin, and quercitrin) were identified and quantified. A fourth quercetin derivative, one dihydrochalcone-related compound, two unknown procyanidins, three hydroxycinnamic derivatives, and two unknown compounds were also found. Among the low-molecular-mass polyphenols analyzed, hydrocaffeic acid was the most abundant compound, representing more than 80% of the total polyphenolic acids. Procyanidins were the most important family among the flavonoid compounds. Discriminant analysis was allowed to correctly classify more than 93% of the ciders, according to the harvest year; the most discriminant variables were an unknown procyanidin and quercitrin.

Sensory and foaming properties of sparkling cider.

The effect of yeast strain and aging time on the chemical composition, analytical, and sensory foam properties of sparkling ciders has been studied. The analytical foam parameters (foamability, HM; Bikerman coefficient, sigma; and foam stability time, T(s)) were significantly influenced by aging and yeast strain. The sensory attributes (initial foam, foam area persistence, bubble size, foam collar, and overall foam quality) improved with aging time. Likewise, the yeast strain positively influenced the assessment of initial foam, foam area persistence, number of bubble chains, and overall foam quality. Significant and positive correlations were found between alcoholic proof, dry extract, total and volatile acidities, total phenols and total proteins, and sigma, whereas HM was negatively correlated with specific gravity, alcoholic proof, dry extract, and total proteins.

Influence of yeast strain and aging time on free amino acid changes in sparkling ciders.

An analytical method for the determination of free amino acids in ciders is reported. It is based on high-performance liquid chromatography with an automatic precolumn derivatization with o-phthaldehyde and 3-mercaptopropionic acid and diode array detection. The method was applied to monitor the amino acids during second fermentation of sparkling ciders. This paper reports the influence of yeast strains and aging time on the amino acid composition of sparkling ciders. The application of principal component analysis enables the ciders to be differentiated on the basis of the two factors considered (yeast strain and aging time). The first principal component, which accounts for 58% of the total variance, achieved the separation according to aging time with serine, glycine, alanine, valine, ornithine, leucine, and lysine as the most important variables. The second principal component, accounting for 28% of the explained variance, is closely related to aspartic acid and asparagine and separates the samples according to the yeast strain.

Typification of cider brandy on the basis of cider used in its manufacture.

A study of typification of cider brandies on the basis of the origin of the raw material used in their manufacture was conducted using chemometric techniques (principal component analysis, linear discriminant analysis, and Bayesian analysis) together with their composition in volatile compounds, as analyzed by gas chromatography with flame ionization to detect the major volatiles and by mass spectrometric to detect the minor ones. Significant principal components computed by a double cross-validation procedure allowed the structure of the database to be visualized as a function of the raw material, that is, cider made from fresh apple juice versus cider made from apple juice concentrate. Feasible and robust discriminant rules were computed and validated by a cross-validation procedure that allowed the authors to classify fresh and concentrate cider brandies, obtaining classification hits of >92%. The most discriminating variables for typifying cider brandies according to their raw material were 1-butanol and ethyl hexanoate.

Application of purge and trap extraction and gas chromatography for determination of minor esters in cider.

The validation of a method based on the purge and trap technique combined with gas chromatography-mass spectrometry-flame ionization detection has been carried out in order to apply it to the analysis of ciders. Although 49 compounds were identified, our work was focused on the study of nine minor esters, obtaining recoveries ranging between 93% for ethyl decanoate and 117% for ethyl 3-methylbutyrate, and a precision (RSDs) ranging between 2.2% for hexyl acetate and ethyl decanoate and 10.9% for isopentyl acetate. To demonstrate the feasibility of the procedure, the method was applied to the analysis of commercial ciders.

Influence of distillation system, oak wood type, and aging time on composition of cider brandy in phenolic and furanic compounds.

A control of phenolic and furanic compounds in cider brandy was carried out during maturation in oak casks, studying three technological factors: distillation (rectification column vs double distillation), oak wood type (French vs American), and aging time (32 months). Gallic acid and benzoic and cinnamic aldehydes significantly increased during maturation of cider brandies, the highest level of these phenolics being obtained when aging was conducted in French oak casks. Benzoic acids increased during aging, though furanic compounds were not influenced by the time factor. Distillation and wood factors significantly influenced furanic concentration; 5-hydroxymethylfurfural not was detected in fresh spirits and was extracted in the highest proportion in French oak. Volatile furanics, such as 5-methylfurfural, furfural, and 2-furylmethyl ketone, were influenced by the distillation factor, with the use of the double distillation system producing a higher level of these compounds. Scopoletin was the majority coumarin detected in cider brandies, the highest yield of which was obtained with the use of American oak.