Occurrence of Isoflavonoids in Brazilian Common Bean Germplasm

Article pubs.acs.org/JAFC

Occurrence of Isoflavonoids in Brazilian Common Bean Germplasm (Phaseolus vulgaris L.) Paula Feliciano de Lima,*,† Carlos Augusto Colombo,‡ Alisson Fernando Chiorato,§ Lydia Fumiko Yamaguchi,∥ Massuo Jorge Kato,∥ and Sérgio Augusto Morais Carbonell§ †

Instituto de Química, Universidade Estadual de Campinas, CP 6154, 13084-971 Campinas, São Paulo, Brazil Centro de Pesquisa e Desenvolvimento de Recursos Genéticos Vegetais and §Centro de Pesquisa e Desenvolvimento de Grãos e Fibras, Instituto Agronômico de Campinas, CP 28, 13012-970 Campinas, São Paulo, Brazil ∥ Núcleo de Apoio à Pesquisas em Diversidade Molecular de Produtos Naturais, Instituto de Química, Universidade de São Paulo, CP 26077, 05508-000 São Paulo, São Paulo, Brazil ‡

ABSTRACT: Common bean (Phaseolus vulgaris) is present in the daily diet of various countries and, as for other legumes, has been investigated for its nutraceutical potential. Thus, 16 genotypes from different gene pools, representing seven types of seed coats and different responses to pathogens and pests, were selected to verify their isoflavone contents. The isoflavonoids daidzein and genistein and the flavonols kaempferol, myricetin, and quercetin were found. Grains of the black type showed the highest concentrations of isoflavonoids and were the only ones to exhibit daidzein. IAC Formoso, with high protein content and source of resistance to anthracnose, showed the greatest concentration of genistein, representing around 11% of the content present in soybean, as well as high levels of kaempferol. Arc 1, Raz 55, and IAC Una genotypes showed high content of coumestrol. The results suggest the use of IAC Formoso to increase the nutraceutical characteristics in common bean. KEYWORDS: Phaseolus vulgaris, flavonoids, isoflavones, functional food, LC-qMS

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INTRODUCTION Phaseolus vulgaris, the common bean, is the most important among the 50 species of native Phaseolus present in the Americas. This species has Mexican origin and contains two major gene pools whose plants differ mainly in relation to seed size: Mesoamerican (small seeds) and Andean (large seeds). The nutraceutical importance of these beans as a protein source is remarkable in several regions or the world, including Brazil.1,2 Studies related to the nutritional and nutraceutical potential of legumes have gained prominence in recent years, especially for soybean and chickpea, since they have higher levels of phytoestrogens such as isoflavonoids.3−5 More recently, common bean has received functional food status as it contains bioactive phenolic compounds and large amounts of complex carbohydrates and fibers, in addition to being a source of iron, phosphorus, magnesium, manganese, zinc, and calcium.3,6,7 However, there is still little information that combines genetic breeding of common bean with its nutraceutical diversity, even in light of a growing number of studies on consumption of the phenolic compounds present in common bean, such as flavonols, isoflavones, and anthocyanins, and their beneficial effects on health. Isoflavonoids have gained prominence because they may play an important role in the prevention and treatment of chronic diseases, as has already been seen in studies that evaluated models in vitro, in vivo, and as interventions in humans. In all these cases, the important biological action of these compounds was observed in antioxidant, “antimutagenic”, and anticarcinogenic activities.3,8−11 A subclass of flavonoids, isoflavonoids, are phytoestrogens that exhibit pseudohormonal properties as a result of their functional and structural similarity to the natural © XXXX American Chemical Society

estrogen 17β-estradiol and may interact with estrogen receptors (ER).12,13 Isoflavone content is variable among plant species and may even vary among genotypes of the same species. In addition, isoflavone content may also be affected by external factors related to crop location, such as temperature, fertilization levels, occurrence of pests and diseases, time since harvest, farming practices, and processing and food preparation methods.14,15 The isoflavone content is significantly less in seeds developed at high temperatures during the grain filling phase, and some studies report a possible relationship between resistance to pathogens and the presence of phytoestrogens, where isoflavoids would act as phytoalexins in plants.16 In the class of coumestans, coumestrol is a metabolite found in different legumes, occurring especially in soybean, alfalfa, clover sprouts, and beans.17 It is discussed mainly in regard to its high affinity for estrogen receptors, around 30−100 times greater than the isoflavonoids exhibiting antagonist functions by competing with ER.18 It also has been shown to decrease ovulation rates, increasing ovarian apoptosis in mammals.19 A novel activity of coumestrol was reported recently and the authors reported that coumestrol binding to human estrogen receptor β inhibited microglia-mediated inflammation.20 In the context of an increasing number of studies on the nutraceutical potential of common bean and the preference of Brazilian consumers for this food, the aim of the present study Received: March 2, 2014 Revised: September 12, 2014 Accepted: September 18, 2014

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dx.doi.org/10.1021/jf5033312 | J. Agric. Food Chem. XXXX, XXX, XXX−XXX

Journal of Agricultural and Food Chemistry

Article

Table 1. Flavonoid Content in 16 Genotypes of Common Bean by LC-qMSa μg/g dry weight of seed flour genotype Arc 1 IAC Diplomata Raz 55 TU IPR Uirapuru IAC Una IAC Alvorada IAC Carioca Comum IAC Formoso IAC Pérola IAC Harmonia IAC Boreal Brancão Argentino IAC Jabola IAC Esperança Flor de Mayo Soja

type of grain

origin

DA

GE

KA

M

Q

CM

B B B B B B C C C C S S W Bo Bo R

M M M M M M M M M M A/M A/M A A/M A/M A

6.82 ± 0.95 nd 8.05 ± 1.83 nd nd 2.30 ± 0.28 nd nd nd nd nd nd nd nd nd nd 62.66 ± 2.68

6.38 ± 0.78 1.10 ± 0.80 5.98 ± 0.89 0.98 ± 0.17 1.45 ± 0.06 2.95 ± 0.13 nd 1.17 ± 0.15 8.93 ± 0.67 0.82 ± 0.65 nd nd nd 1.39 ± 0.09 2.16 ± 0.29 1.93 ± 0.33 77.80 ± 8.38

2.28 ± 0.19 0.78 ± 0.06 5.40 ± 0.78 4.85 ± 3.53 2.82 ± 0.39 1.58 ± 0.74 1.64 ± 1.18 35.09 ± 2.55 60.07 ± 3.17 16.37 ± 11.53 0.40 ± 0.32 2.69 ± 0.40 nd 117.65 ± 5.09 72.13 ± 1.06 6.47 ± 0.59 nd

nd nd 26.15 ± 22.86 78.94 ± 13.94 nd nd nd nd nd nd nd nd nd nd nd nd nd

2.56 ± 0.44 1.09 ± 0.17 8.34 ± 0.91 15.85 ± 3.57 4.37 ± 3.80 2.80 ± 0.94 nd nd 9.88 ± 3.04 nd nd nd nd 2.79 ± 0.65 nd nd nd

7.75 ± 0.33 2.32 ± 0.09 17.67 ± 4.40 0.75 ± 0.14 2.60 ± 0.21 5.95 ± 0.40 nd 1.19 ± 0.16 16.85 ± 0.81 3.31 ± 2.87 nd nd nd 4.50 ± 1.04 2.53 ± 0.36 nd nd

DA + GE 13.20 14.03

5.25

140.46

a

Values shown are means of three repetitions. Type of grain: B, black; C, carioca; S, striped; W, white; Bo, bolinha; R, red. Origin: A, Andean; M, Mesoamerican. Isoflavonoids: DA, daidzein, and GE, genistein. Flavonols: KA, kaempferol, M, myricetin, and Q, quercetin. CM, coumestrol. nd, not detected.

was to assess the flavonoid and coumestrol content in germplasm with a broad genetic base of the species, for the purpose of introducing new nutraceutical parameters in the crop breeding program.

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MATERIALS AND METHODS

Plant Material. The study was carried out at the Campinas (SP) (22° 54′ 20″ S, 47° 03′ 39″ W) during the rainy season (December− May) with 16 common bean genotypes from the two major gene pools with different seed coats and grain sizes, and one soybean genotype used as a reference (Table 1, Figure 1). Seeds of each cultivar were sown in a 5-m-length row (10 plants/m) for a total area of 105 m2. After 3 months, grains from all genotypes were collected and kept in an air-circulation laboratory oven at 30 °C for drying to constant weight, and afterward ground in a mill and stored with protection against moisture. Standards. Authentic standard daidzein, daidzin, genistein, and genistin (isoflavonoids); quercetin, kaempferol, and myricetin (flavonols); coumestrol (coumestan); and chrysin were purchased from Sigma Chemical Co. (St. Louis, MO). Stock solutions were prepared by dissolving the standards in MeOH−H2O (1:1) for a final concentration of 10 mg/mL. Solvents. All solvents used were HPLC-grade and were purchased from Sigma Chemical Co. (St. Louis, MO). Extraction and Purification of Flavonoids. Portions (2 g) of dry and ground grains were extracted with 60 mL of 70% ethanol (pH 2.0, adjusted with formic acid) for 24 h at ambient temperature.21 The extracts were partitioned with 60 mL of hexane, and the aqueous phase was evaporated under reduced pressure to dryness. The residue of each extract was resuspended in MeOH−H2O (1:1), filtered through a 13 mm PTFE membrane filter of 0.45 μm, and kept at −20 °C up to the time of analysis. Each extract was prepared and analyzed in triplicate. Identification and Quantification of Flavonoids. Quantification of all compounds was conducted from construction of a calibration curve with chrysin as internal standard. Individual identification of the flavonoids (negative mode) was performed on the basis of retention times, spectroscopic data, and mass-to-charge ratio (m/z) of each standard by use of 10 points of concentration, 0.005−5 μg/mL, containing the internal standard at a final

Figure 1. Seed coat colors of the 16 genotypes evaluated. (A) Arc1, (B) IAC Diplomata, (C) Raz 55, (D) TU, (E) IPR Uirapuru, (F) IAC Una, (G) IAC Alvorada, (H) IAC Carioca Comum, (I) IAC Formoso, (J) IAC Pérola, (K) IAC Harmonia, (L) IAC Boreal, (M) Brancão Argentino, (N) IAC Jabola, (O) IAC Esperança, and (P) Flor de Mayo. concentration of 4 μg/mL (Table 2). Results were expressed as milligrams per gram dry weight of seed flour. Analytical Techniques and Equipment. The LC-ESI-QTOFMS MicroTOF-QII Bruker equipped with an autosampler, binary pump, thermostated column compartment, and multiple-wavelength detector was used. Separations of flavonoids were achieved on a 150 × 2.0 mm i.d, 5 μm C18 Luna column (Phenomenex, Torrence, CA) at 30 °C. Injection volume was 20 μL at a flow rate of 0.25 mL/min. Linear gradient elution was performed with solvent A (methanol/ formic acid, 95:5 v/v) and solvent B (5% formic acid in water, v/v) at B



Article

With the exceptions of IAC Alvorada (carioca seed coat), IAC Boreal and IAC Harmonia (striped seed coat), and Brancão Argentino (white seed coat), all the genotypes exhibited genistein (GE), ranging from 0.82 (IAC Pérola) to 8.93 (IAC Formoso) μg/g. Among the types of grains, the genotypes of black seed coat and Mesoamerican origin stood out: in addition to containing GE, they were the only ones to also exhibit daidzein (DA), as in the case of IAC Una (2.30 ± 0.28 μg/g), Arc 1 (6.82 ± 0.95 μg/g), and Raz 55 (8.05 ± 1.83 μg/g). A similar result was reported by Romani et al.,26 who observed the presence of ́ these isoflavonoids in a local variety of black bean, and by DiazBatalla et al.25 in assessment of cultivated and wild Mexican common beans. Liggins et al.10 also reported levels of daidzein and genistein in beans with red seed coat. IAC Una, Arc 1, and Raz 55 were the only ones to show DA and GE at higher levels. IAC Una is considered to be the main Brazilian cultivar for resistance to the anthracnose pathogen (Colletotrichum lindemuthianum).27 In Arc 1 and Raz55, the presence of the protein arcelin was observed, which is considered to be the main source of resistance to Zabrotes subfasciatus, one of the main pests of stored grains that occurs in common bean.28 With variation from 0.82 to 14.03 μg/g isoflavonoids for the genotypes assessed, the results obtained were similar to those reported in common bean, where contents from 6 to 14 μg/g were found for different landraces, from 8.3 to 13.4 μg/g for striped common beans, and from 7.1 to 11.4 μg/kg of isoflavonoids (daidzein and genistein) in haricot bean.26,27,29 In soybean, a microbial toxic action of the glycosides daidzein and genistein was proposed in resistance to fungus infection of seeds and seedlings.30 In the genus Phaseolus, Adesanya et al.31 tested isoflavones, isoflavonones, pterocarpans, and coumestan from five species against Aspergillus niger and Cladosporium cucumericum and observed the importance of at least a phenolic function to activate the fungitoxic activity of these compounds, making assessment of the structure−fungitoxic activity relationships within these groups of isoflavonoids possible. Corroborating the results found by Beninger et al.,32 flavonols occurred in all the genotypes assessed, except for the genotype Brancão Argentino (Table 1). The highest occurrence of the flavonol kaempferol (KA) was seen in grains with a light-colored seed coat like those of the bolinha and carioca types. IAC Jabola and IAC Esperança, both with bolinha and carioca seed coat types, such as IAC Carioca Comum, IAC Formoso, and IAC Pérola (all of Mesoamerican origin), exhibited kaempferol contents that ranged from 16.37 to 117.65 μg/g (Table 1). A similar profile was found by Ranilla et al.6 in assessment of 25 Brazilian common bean cultivars as a function of color of the seed coat and of the cotyledon. This flavonol modifies a series of cell signaling pathways acting with less toxicity in comparison to standard chemotherapies.33 Quercetin (Q) and myricetin (M) occurred preferentially in black grains, ranging from 1.09 to 15.85 μg/g and from 26.15 to ́ 78.94 μg/g, respectively. Diaz-Batalla et al.25 found a similar occurrence of quercetin in black grains and kaempferol in lighter-colored grains. In fact, quercetin and kaempferol are the most abundant flavonols in foods and are antioxidant and chelating compounds with beneficial effects on health.34 They are also associated with the function of protecting seeds against pathogens and predators.35 An explanation for the fact of common bean being considered a source of flavonols was the high occurrence of quercetin and kaempferol in the cultivar

Table 2. LC-qMS and Calibration Parameters of Flavonoids compd

tR (min)

[M − H]− (m/z)

r2

concn (μM)

daidzein genistein quercetin kaempferol myricetin coumestrol chrysin

13.39 13.39 15.56 17.47 13.10 18.19 20.25

253 269 301 285 317 267 253

0.976 0.991 0.996 0.995 0.999 0.991

0.02−19.7 0.02−18.5 0.02−16.5 0.02−17.5 0.01−15.7 0.02−18.7 0.02−19.7

30% A/70% B from 0 to 4 min, 100% A from 4 to 50 min, and 30% A/ 70% B from 50 to 65 min. Chromatograms were recorded by UV detection at 254, 270, and 330 nm and by ESI-MS in negative-ion scan mode (100−600 m/z) with nitrogen as nebulizer gas at 58 psi and 4.5 kV capillary voltage at 350 °C. Identification of flavonoids and coumestan was based on the following ions ([M − H]−, m/z) for each analyte: 253 for daidzein, 269 for genistein, 415 for daidzin, 431 for genistin, 301 for quercetin, 285 for kaempferol, 317 for myricetin, 267 for coumestrol, and 253 for chrysin. Data Analysis. Descriptive statistics were used to characterize the data overall and across factors. To identify statistical differences in flavonoid content among factors, ANOVA and canonical analysis methods were employed. Statistical analyses were performed by use of the Genes program.22

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RESULTS AND DISCUSSION Identification of Bean Flavonoids and Coumestrol. The maximum and minimum temperatures of the location where the plant material was grown ranged from 17.6 to 27.5 °C (Figure 2) with mean monthly rainfall of 7.14 mm

Figure 2. Average maximum and minimum temperature during the growth biological assay.

(December 2010−May 2011). Studies on genetic and environmental effects, performed on soybean in locations where mean temperatures ranged from 19 to 27 °C, manifested a strong influence of the environment on isoflavonoid contents.23,24 In all 16 genotypes assessed, we found the flavonoids quercetin (Q), myricetin (M), and kaempferol (KA) and the nonglycosylated forms of isoflavonoids daidzein (DA) and genistein (GE), both of greater occurrence among the legumes, as well as coumestrol, all with wide variation of occurrence among the genotypes of the study (Table 1).25 C



Article

IAC Formoso, exhibiting contents from 9.88 ± 3.04 to 60.07 ± 3.17 μg/g.11 Genotypes of Mesoamerican origin with black and carioca seed coats and of Andean origin with bolinha seed coat manifested coumestrol (Table 1). Coumestrol is an important compound which contains high levels of estrogenic activity, 30−100 times greater than that of the isoflavones, and it was identified in all the black grains (Table 1), ranging from 0.75 (TU) to 17.67 (Raz 55) μg/g.18 Among the grains with carioca seed coat, IAC Formoso presented the greatest content of this coumestan (16.85 ± 0.81 μg/g). Konar36 assessed common bean and lentils through LC-MS/MS and found this coumestan in common bean (18.5 μg/kg) and a mean value of 16.9 μg/kg in yellow, green, and red lentils. Moreover, by LC-MS/MS and with standards marked with 13C, Kuhnle et al.37 found contents ranging from 1 to 10 μg/100 g for common bean, values considered very inferior to ours, even when it is taken into account that the authors expressed their results by fresh weight. IAC Formoso exhibited the greatest concentration of genistein among all the genotypes assessed (8.93 ± 0.67 μg/ g), around 11% of the total presented by the soybean control. Genistein is considered to be the isoflavone with greatest biological activity and is furthermore classified as an inducible phytoalexin.21 In addition, IAC Formoso exhibited the second greatest concentration of coumestrol (16.85 ± 0.81 μg/g), a compound that has been reported as an estrogen receptor twice as powerful as genistein and, furthermore, a modulator in hormone production of the thymus.38 Grouping Common Bean According to Seed Coat Color. Canonical variable analysis (CVA) was applied to the values of flavonoid content of the genotypes assessed. This multivariate analysis is similar to principal component analysis, and it is especially used in discriminating analyses with replication of observations because it is based on the variances of these replications. The first three canonical variables explained 97.5% of the total variance among the 16 genotypes and showed the formation of three groups in accordance with the flavonoid and coumestrol content (Figure 3). A first group gathered the genotypes Arc 1 and Raz 55, both with a black seed coat and of Mesoamerican origin and with the greatest isoflavonoid contents found in the study. A second group was composed of IAC Jabola and IAC Esperança, both with bolinha type grains and the greatest concentrations of the flavonol kaempferol, which occurred preferentially in light-colored grains. All the other genotypes were in a third group. Canonical variable 1 (VC1), with 78.8% of the total variance, differentiated the genotypes of the study in accordance with daidzein and genistein content. Thus, Arc 1 and Raz 55 were differentiated from IAC Jabola and IAC Esperança, by presenting the highest and lowest contents of daidzein and genistein, respectively (Table 1). Canonical variable 2 (VC2) represents 13.2% of the variation and showed the separation of the genotypes in two groups with contrasting values of concentration of total flavonoids. In VC2, the first group gathered IAC Carioca comum, IAC Jabola, IAC Esperança, IAC Formoso, IAC Una, TU, Arc 1 and Raz 55. A second group was formed by IAC Diplomata, IPR Uirapuru, IAC Alvorada, IAC Pérola, IAC Harmonia, IAC Boreal, Flor de Mayo, and Brancão Argentino. The variability found for flavonoid content in the grains of 16 genotypes assessed under field conditions may be explained by how these materials arose. Thus, factors such as seed coat type

Figure 3. Multivariate exploratory analysis for 16 genotypes of common bean. Three groups were formed after canonical analysis: the yellow group contained IAC Diplomata (1), IAC Una (2), IPR Uirapuru (3), IAC Alvorada (4), IAC Pérola (5), IAC Carioca comum (6), TU (7), IAC Harmonia (8), IAC Boreal (9), Brancão Argentino (10), Flor de Mayo (13) and IAC Formoso (16) genotypes; The red group contained IAC Jabola (11) and IAC Esperança (12) genotypes; and the green group contained Arc1 (14) and Raz 55 (15) genotypes.

and selection for tolerance to biotic and abiotic stresses may explain the different concentrations of metabolites found. Related studies reinforce this hypothesis, where differences in the variability of isoflavonoids and flavonoids in accordance with the type of seed coat, genotype, and genotype− environmental interaction were cited by various authors.23,39,40 Important levels of flavonoids were observed in the germplasm of common bean grown under the same environmental conditions and high variability between groups with different seed coat colors. Among the beans with carioca-type seed coat, which represents nearly all of the common beans grown, IAC Formoso, a cultivar recently introduced for planting on a commercial scale, showed high values of flavonoids and coumestrol. This result suggests that this cultivar may be used in the genealogy of future genetic materials with a view toward adoption of nutraceutical characteristics in common bean.

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AUTHOR INFORMATION

Corresponding Author

*Telephone (+5519) 35213105; fax (+5519) 35213023; e-mail [email protected]. Notes

The authors declare no competing financial interest.

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ACKNOWLEDGMENTS This work was financially supported by FAPESP. C.A.C., A.F.C., M.J.K., and S.A.M.C. thank the Conselho Nacional de ́ Desenvolvimento Cientifico e Tecnológico (CNPq) for granting fellowships. D



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and domestic wastewater sewage treatment plants. Anal. Chim. Acta 2005, 531, 229−237. (19) Lee, Y.-H.; Yuk, H. J.; Park, K.-H.; Bae, Y.-S. Coumestrol induces senescence through protein kinase CKII inhibition-mediated reactive oxygen species production in human breast cancer and colon cancer cells. Food Chem. 2013, 141, 381−388. (20) Saijo, K.; Collier, J. G.; Li, A. C.; Katzenellenbogen, J. A.; Glass, C. K. An ADIOL-ERbeta-CtBP trans repression pathway negatively regulates microglia-mediated inflammation. Cell 2011, 145, 584−595. (21) Romani, A.; Vignolini, P.; Galardi, C.; Aroldi, C.; Vazzana, C.; Heimler, D. Polyphenolic content in different plant parts of soy cultivars grown under natural conditions. J. Agric. Food Chem. 2003, 51, 5301−5306. (22) Cruz, C. D. Programa Genes - Análise multivariada e simulação. 1st ed.; Editora UFV, Universidade Federal de Viçosa: Viçosa, Brazil, 2006; Vol. 1. (23) Carrão-Panizzi, M. C.; Del Pino Beleia, A.; Kitamura, K.; Oliveira, M. C. N. Effects of genetics and environment on isoflavone content of soybean from different regions of Brazil. Pesqui. Agropecu. Bras. 1999, 34, 1787−1795. (24) Tsukamoto, C.; Shimada, S.; Igita, K.; Kudou, S.; Kokubun, M.; Okubo, K.; Kitamura, K. Factors affecting isoflavone content in soybean seeds: Changes in isoflavones, saponins, and composition of fatty acids at different temperatures during seed development. J. Agric. Food Chem. 1995, 43, 1184−1192. (25) Díaz-Batalla, L.; Widholm, J. M.; Fahey, G. C.; CostanoTostado, E.; Paredez-López, O. Chemical components with health implications in wild and cultivated Mexican common bean seeds (Phaseolus vulgaris L.). J. Agric. Food Chem. 2006, 54, 2045−2052. (26) Romani, A.; Vignolini, P.; Galardi, C.; Mulinacci, N.; Benedettelli, S.; Heimler, D. Germplasm characterization of Zolfino landraces (Phaseolus vulgaris L.) by flavonoid content. J. Agric. Food Chem. 2004, 52, 3838−3842. (27) Carbonell, S. A. M.; Ito, M. F.; Pompeu, A. S.; Francisco, F. G.; Ravagnani, S.; Almeida, A. L. L. Raças fisiológicas de Colletotrichum lindemuthianum e reaçaõ de linhagens e cultivares de feijoeiro no Estado de São Paulo. Fitopatol. Bras. 1999, 24, 60−65. (28) Ribeiro-Costa, C. S.; Pereira, P. R. V. S.; Zulovski, L. Desenvolvimento de Zabrotes subfasciatus (Boh.) (Coleoptera: Chrysomelidae, Bruchinae) em genótipos de Phaseolus vulgaris L. (Fabaceae) cultivados no estado do Paraná e contendo arcelina. Neotrop. Entomol. 2007, 36, 560−564. (29) Konar, N.; Poyrazoglu, E. S.; Demir, K.; Artik, N. Determination of conjugated and free isoflavones in some legumes by LC-MS/MS. J. Food Compos. Anal. 2012, 25, 173−178. (30) Graham, T. L.; Kim, J. E.; Graham, M. Y. Role of constitutive isoflavone conjugates in the accumulation of glyceollin in soybean infected with Phytophthora megasperma. Mol. Plant-Microbe Interact. 1990, 3, 157−166. (31) Adesanya, S. A.; O’Neill, M. J.; Roberts, M. F. Structure-related fungitoxicity of isoflavonoids. Physiol. Mol. Plant Pathol. 1986, 29, 95− 103. (32) Beninger, C. W.; Hosfield, G. L.; Bassett, M. J. Flavonoid composition of three genotypes of dry bean (Phaseolus vulgaris) differing in seed coat color. J. Am. Hortic. Sci. 1999, 124, 514−518. (33) Zhang, Y.; Chen, A. Y.; Li, M.; Chen, C.; Yao, Q. Ginkgo biloba extract kaempferol inhibits cell proliferation and induces apoptosis in pancreatic cancer cells. J. Surg. Res. 2008, 148, 17−23. (34) Heim, K. E.; Tagliaferro, A. R.; Bobilya, D. J. Flavonoid antioxidants: Chemistry, metabolism and structure-activity relationships. J. Nutri. Biochem. 2002, 13, 572−584. (35) Islam, F. M. A.; Rengifo, J.; Redden, R. J.; Basford, K. E.; Beebe, S. E. Association between seed coat polyphenolics (tannins) and disease resistance in common bean. Plant Food Hum. Nutr. 2003, 58, 285−297. (36) Konar, N. Non-isoflavone phytoestrogenic compound contents of various legumes. Eur. Food Res. Technol. 2013, 236, 523−530. (37) Kuhnle, G. G. C.; Dell’Aquila, C.; Aspinall, S. M.; Runswick, S. A.; Joosen, A. M. C. P.; Mulligan, A. A.; Bingham, S. A. Phytoestrogen

ABBREVIATIONS ANOVA, analysis of variance; DA, daidzein; GE, genistein; HPLC, high-performance liquid chromatography; KA, kaempferol; LC-ESI-QTOF-MS, liquid chromatography−electrospray ionization quadrupole time-of-flight mass spectrometry; M, myricetin; PTFE, poly(tetrafluoroethylene); Q, quercetin

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REFERENCES

(1) Angioi, S. A.; Rau, D.; Attene, G.; Nanni, L.; Bellucci, E.; Logozzo, G.; Negri, V.; Spagnoletti-Zeuli, P. L.; Papa, R. Beans in Europe: Origin and structure of the European landraces of Phaseolus vulgaris L. Theor. Appl. Genet. 2010, 121, 829−843. (2) Carbonell, S. A. M.; Chiorato, A. F.; Carvalho, C. R. L.; Júnior, E. U. R.; Ito, M. A.; Borges, W. L. B.; Ticelli, M.; Santos, N. C. B.; Gallo, P. B. IAC Formoso: New carioca common bean cultivar. Crop Breed. Appl. Biotechnol. 2010, 10, 374−376. (3) Espinosa-Alonso, L. G.; Lygin, A.; Widholm, J. M.; Valverde, M. E.; Paredes-Lopez, O. Polyphenols in wild and weedy Mexican common beans (Phaseolus vulgaris L.). J. Agric. Food Chem. 2006, 54, 4436−4444. (4) Boniglia, C.; Carratú, B.; Gargiulo, R.; Giammarioli, S.; Mosca, M.; Sanzini, E. Content of phytoestrogens in soy-based dietary supplements. Food Chem. 2009, 115, 1389−1392. (5) Kuhnle, G. G. C.; Dell’Aquila, C.; Runswick, S. A.; Bingham, S. A. Variability of phytoestrogen content in foods from different sources. Food Chem. 2009, 113, 1184−1187. (6) Ranilla, L. G.; Genovese, M. I.; Lajolo, F. M. Polyphenols and antioxidant capacity of seed coat and cotyledon from Brazilian and Peruvian bean cultivars (Phaseolus vulgaris L.). J. Agric. Food Chem. 2007, 55, 90−98. (7) Broughton, W. J.; Hernández, G.; Blair, M. W.; Beebe, S. Beans (Phaseolus spp.) - Model food legumes. Plant Soil 2003, 252, 55−128. (8) Dong, M.; He, X.; Liu, R. H. Phytochemicals of black bean seed coats: Isolation, structure elucidation, and their antiproliferative and antioxidative activities. J. Agric. Food Chem. 2007, 55, 6044−6051. (9) Pascual-Teresa, S.; Hallundb, J.; Talbotc, D.; Schrootd, J.; Williamse, C. M.; Bugelb, S.; Cassidy, A. Absorption of isoflavones in humans: effects of food matrix and processing. J. Nutr. Biochem. 2006, 17, 257−264. (10) Liggins, L.; Bluck, L. J. C.; Runswick, S.; Atkinson, C.; Coward, W. A.; Bingham, S. A. Daidzein and genistein contents of vegetables. Br. J. Nutr. 2000, 84, 717−725. (11) Dinelli, G.; Bonetti, A.; Minelli, M.; Marotti, I.; Catizone, P.; Mazzanti, A. Content of flavonols in Italian bean (Phaseolus vulgaris L.) ecotypes. Food Chem. 2006, 99, 105−114. (12) Kuhnle, G. G. C.; Dell’Aquila, C.; Low, Y. L.; Kussmaul, M.; Bingham, S. A. Extraction and quantification of phytoestrogens in foods using automated solid-phase extraction and LC/MS/MS. Anal. Chem. 2007, 79, 9234−9239. (13) Boué, S. M.; Burow, M. E.; Wiese, T. E.; Shih, B. Y.; Elliott, S.; Carter-Wientjes, C. H.; McLachlan, J. A.; Bhatnagar, D. Estrogenic and antiestrogenic activities of phytoalexins from red kidney bean (Phaseolus vulgaris L.). J. Agric. Food Chem. 2004, 59, 112−120. (14) Wang, H.; Murphy, P. A. Isoflavone content in commecial soybean foods. J. Agric. Food Chem. 1994, 42, 1666−1673. (15) Eldridge, A. C.; Kwolek, W. F. Soybean isoflavones: Effect of environment and variety on composition. J. Agric. Food Chem. 1983, 31, 394−396. (16) Morris, P. F.; Savard, M. E.; Ward, E. W. B. Identification and accumulation of isoflavonoids and isoflavone glucosides in soybean leaves and hypocotyls in resistance responses to Phytophthora megasperma f.sp. glycinea. Physiol. Mol. Plant Pathol. 1991, 39, 229− 244. (17) Hong, Y. H.; Wang, S. C.; Hsu, C.; Lin, B. F.; Kuo, Y. H.; Huang, C. J. Phytoestrogenic compounds in alfalfa sprout (Medicago sativa) beyond coumestrol. J. Agric. Food Chem. 2011, 59, 131−137. (18) Bacaloni, A.; Cavaliere, C.; Faberi, A.; Foglia, P.; Samperi, R.; Lagana, A. Determination of isoflavones and coumestrol in river water E



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content of fruits and vegetables commonly consumed in the UK based on LC-MS and 13C-labelled standards. Food Chem. 2009, 116, 542− 554. (38) Ndebele, K.; Graham, B.; Tchouwou, P. B. Estrogenic activity of coumestrol, DDT, and TCDD in human cervical cancer cells. Int. J. Environ. Res. Public. Health 2010, 7, 2045−2056. (39) Lee, S. J.; Seguin, P.; Kim, J. J.; Moond, H. I.; Ro, H. M.; Kim, E. H.; Seo, S. H.; Kang, E. Y.; Ahn, J. K.; Chung, I. M. Isoflavones in Korean soybeans differing in seed coat and cotyledon color. J. Food Compos. Anal. 2010, 23, 160−165. (40) Kim, J. K.; Kim, E. H.; Lee, O. K.; Park, S. Y.; Lee, B.; Kim, S. H.; Park, I.; Chung, I. M. Variation and correlation analysis of phenolic compounds in mung bean (Vigna radiata L.) varieties. Food Chem. 2013, 141, 2988−2997.

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Occurrence of Isoflavonoids in Brazilian Common Bean Germplasm

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