(Microsoft Word - 11353 NSB Ch\341vez-Mendoza 2023.03.17.docx) Received: 20 Sep 2022. Received in revised form: 08 Mar 2023. Accepted: 15 Mar 2023. Published online: 17 Mar 2023. From Volume 13, Issue 1, 2021, Notulae Scientia Biologicae journal uses article numbers in place of the traditional method of continuous pagination through the volume. The journal will continue to appear quarterly, as before, with four annual numbers. SHSTSHSTSHSTSHST Horticulture and ForestryHorticulture and ForestryHorticulture and ForestryHorticulture and Forestry Society of TransylvaniaSociety of TransylvaniaSociety of TransylvaniaSociety of Transylvania Chávez-Mendoza C and Sánchez E (2023) Notulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia Biologicae Volume 15, Issue 1, Article number 11353 DOI:10.15835/nsb15111353 Research ArticleResearch ArticleResearch ArticleResearch Article.... NSBNSBNSBNSB Notulae Scientia Notulae Scientia Notulae Scientia Notulae Scientia BiologicaeBiologicaeBiologicaeBiologicae Antioxidant capacity and nutraceutical compounds content of six Antioxidant capacity and nutraceutical compounds content of six Antioxidant capacity and nutraceutical compounds content of six Antioxidant capacity and nutraceutical compounds content of six common bean (common bean (common bean (common bean (Phaseolus vulgaris Phaseolus vulgaris Phaseolus vulgaris Phaseolus vulgaris L.L.L.L.) varieties ) varieties ) varieties ) varieties harvested in Morelos, Mexicoharvested in Morelos, Mexicoharvested in Morelos, Mexicoharvested in Morelos, Mexico Celia CHÁVEZ-MENDOZA, Esteban SÁNCHEZ* Centro de Investigación en Alimentación y Desarrollo A. C. Av. Cuarta Sur #3820. Fracc. Vencedores del Desierto, C.P. 33089, Delicias, Chihuahua, México; celia.chavez@ciad.mx; esteban@ciad.mx (*corresponding author) AbstractAbstractAbstractAbstract Common bean is considered one of the most important legumes in the world. It is the main source of protein, calories, B vitamins, minerals, polyphenols and other elements, which collectively give it a high nutraceutical value. In Mexico a great agrobiodiversity exists in the production of this grain, which implies the need to generate information regarding its nutritional quality as a tool to apply future genetic improvement programs. The purpose of this study was to characterize the antioxidant capacity (AC) and nutraceutical content of six bean varieties produced in Morelos State, Mexico. Grain morphometric characteristics, color (L, a*, b* chroma and °hue), nutritional quality, AC (DPPH), nutraceutical compounds content, micro and macro nutrients were determined. A significant effect (p < 0.05) of variety on almost all the variables evaluated except for phytic acid, P, K, Ca, C, S and H was observed. Lower lightness was obtained in varieties with darker colors such as ‘Negro’/102 (24.96), ‘Negro’/104 (26.85) and ‘Sangre de Toro’ (32.41) and higher lightness in lighter colored varieties such as ‘Peruano’ bean (69.21), ‘Pinto’ (65.94) and ‘Flor de Mayo’ (50.14). Nutritional and nutraceutical quality of the latter genotype stood out, as it had the highest crude fiber content (5.71 %), total phenols (4.24 mg GAE g-1), flavonoids (1.99 mg CE g-1), AC (96.76% Inhibition), and a high protein content (23.29%). Results also exhibited significant correlation (p < 0.05) between total phenols and flavonoids with AC. It is concluded that the nutritional characterization carried out on bean varieties from important producing areas in Mexico provide a valuable database for genotype selection with high functional and nutritional character, either to be grown for direct consumption, future biofortification or breeding programs. Keywords:Keywords:Keywords:Keywords: antioxidant capacity; bioactive compounds; micronutrients; Mexican bean; Morelos IntroductionIntroductionIntroductionIntroduction Nutraceutical products are defined as chemical or biological substances that can be found as natural components of food or added to it, and that are particularly beneficial, both in disease prevention and in improvements in physiological functions of the organism. The consumer's and the general population's interest in obtaining optimal diets to maintain good health and prolong lifespan has led to an increase in natural food markets in which this type of product has priority (Pérez, 2006). Common bean is considered one of the most important legumes in the world. It is the main source of protein, calories, B vitamins, minerals, polyphenols https://www.notulaebiologicae.ro/index.php/nsb/index Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 2 and other elements, which collectively give it a high nutraceutical value (Herrera-Hernández et al., 2018). This legume is considered a functional food and has gained great interest for study due to its high content of bioactive compounds such as enzyme inhibitors, lectins, phytates, oligosaccharides and phenolic compounds that may have metabolic roles in humans and animals that consume them frequently. Its antioxidant capacity, antimutagenic and antiproliferative effect have been associated with the presence of phenolic compounds, and this could explain the numerous scientific studies that have suggested that the consumption of this food is related to several beneficial health effects, such as reduction of coronary heart disease, protective effects against cancer, and decrease of diabetes and obesity risk (Gálvez et al., 2007). The antioxidant activity of beans is due to the presence of phenolic acids and flavonoids, mainly tannins, both raw and cooked, gallic, vanillic, p- coumaric, ferulic, sinapic and chlorogenic acid, which are of great importance as precursors in the synthesis of phenolic compounds in plants (Huber et al., 2016). Legumes are also an excellent source of micronutrients. They are a source of Se, thiamine, niacin, folate, riboflavin and pyridoxine. They also contain vitamin E and A as well as Fe and Zn, although the Fe content can vary greatly depending on the variety, for example, white beans contain almost twice as much Fe as black beans. However, most of the Fe contained is tightly bound to phytates, which reduce absorption and may contribute to Fe deficiencies in countries where beans and other legumes are a staple food (Mudryj et al., 2014). Nevertheless, beans have the potential to treat Fe deficiency anemia and other diseases associated with micronutrient deficiencies that affect a large number of people around the world. In this regard, biofortification of this crop is a technique that has been launched with the rationale that high mineral content grains will increase the supply and availability of non-heme Fe in various human populations. Such nutritional improvement focuses on both increasing nutrient content and reducing the anti-nutritional factors contained in the plant, such as oxalates, phytates and tannins, which together affect the bioavailability of these nutrients to consumers (Diaz et al., 2010). Common beans are a staple food in many Latin American and African countries. Mexico is considered the center of origin of this legume, which has been consumed since pre-Hispanic era (Espinosa-Alonso et al., 2006). This country has the widest variety of beans and is accepted as the center of origin of the common bean, since 47 out of 52 species classified in the Phaseolus genus were identified in Mexico (Silva-Cristobal et al., 2010). In Mexico, there is a great diversity of this crop, in which color is one of the attributes that determine consumption preferences in different Mexican regions, such as yellow in the northwest, beige with brown and cream spots in the northeast, black in the south and various specific colors in the central region (Espinosa- Alonso et al., 2006). It is known that the coat color is attributed to the presence and quantity of polyphenols such as flavonol glycosides, condensed tannins and anthocyanins which function to protect the seed against predators and pathogens (Beninger et al., 1999; Takeoka et al., 1997). Since there are several factors that influence the nutritional quality of the bean grain, and due to the wide agrobiodiversity that exists in Mexico, the objective of this study was to characterize the antioxidant capacity and nutraceutical content of six bean varieties from Morelos, Mexico, in order to select varieties that have the potential to be bio-fortified with micronutrients, with a focus on improving bean nutritional quality, thus contributing to the nutrition of the population, especially the most unprotected whose food base is legumes. Materials and MethodsMaterials and MethodsMaterials and MethodsMaterials and Methods Sample preparation Six common bean varieties representative of Morelos State, Mexico, were used for the study were used, which are presented in Table 1. Seeds were harvested in 2018 and collected and analysed in the same year. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 3 Table 1.Table 1.Table 1.Table 1. Bean varieties grown in Morelos State, Mexico used in the study Variety Origin Laboratory work classification number Color Photography ‘Sangre de Toro’ Tlayacapan Morelos 98 Red ‘Peruano’ Cuautla Morelos 100 Yellow ‘Flor de Mayo’ Cuautla Morelos 101 Spotted ‘Negro’ Cuautla Morelos 102 Black ‘Pinto’ Cuautla Morelos 103 Pinto ‘Negro’ Cuautla Morelos 104 Black For the analysis, 100 grains of each variety were used. Samples were ground to a fine powder which was stored in polyethylene bags and kept in a desiccator until analysis. The determinations were made in triplicate. Morphometric characteristics determination of beans Seed weight. One hundred seeds of each bean variety were placed in a Petri dish and weighted using an analytical scale (And Company Limited, Milpitas, CA, U.S.A.). The result was reported in g of 100 seeds. Length, width and thickness. These were determined using a digital vernier (Steren®, Azcapotzalco, Mexico City, Mexico). One hundred seeds of each variety studied were used for the test. Results were expressed in mm. Color determination For color evaluation, 100 seeds of each variety were taken and placed in a glass petri dish until the container was full, and using a portable colorimeter Konica Minolta DP-400 (Minolta Co. Ltd. Osaka, Japan) the CIELAB system color coordinates (L*, a* and b*) were obtained from the surface of these samples. Where parameter L* represents brightness, which varies from 0 (black) to 100 (white); a* can have either positive (red) or negative (green) values; and b* represents yellow when the value is positive and blue when it is negative. Color coordinates L*, a* and b* were used to obtain the CIEL*C*hº color space, where C represents chroma or color saturation and hº is the hue angle or hue representing color according to the angle on the 360 Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 4 ° color wheel, with red-purple at 0 °, yellow at 90 °, blue-green at 180 ° and blue at 270°, counter clockwise (McGuire, 1992). Chroma and hue were calculated using the following formulas (McGuire, 1992): �∗ = (�∗2 + �∗2)1/2 (1) °h= 360° + [(arctan (b*/a*)) /6.2832]*360 (2) When a*>0 and b*<0 Or °h= [(arctan (b*/a*)) /6.2832]*360 (3) When a>0 y b>0 Or °h= 180 + [(arctan (b*/a*)) /6.2832]*360 (4) When a<0 y b<0 Nutritional quality analysis of bean Ash content determination Ash analysis was performed using the method proposed by Mexican Norm NMX-F-F066-S-1978 (1978). One g of sample was weighed in a crucible and kept at constant weight (100 ºC for 2 h) then placed in a desiccator and taken to a muffle (Felisa®) where it was kept at 600 ºC to carbonize the sample until calcination was reached. Results were expressed as percentage of ash. Fat content determination Fat content in the bean samples was determined using the Golfish method proposed by the Association of Official Analytical Chemist (AOAC) (2000). Goldfish flasks were dried and kept to constant weight in an oven. The Goldfish Labconco® grease extractor was then assembled and the sample was placed inside filter paper and covered with absorbent cotton, to be introduced into the equipment. The solvent (petroleum ether) was added and kept under reflux for 2.5 h. After the extraction time was over, the solvent was recovered by distillation, retaining only the fat. Finally, the flask with fat was weighed and expressed as a percentage. Moisture content determination Moisture was obtained by the open-capsule drying method proposed by the AOAC (2000). For the analysis, 1 g of sample was taken from each bean variety, which was weighed in an aluminium capsule that was previously dried at 75 ºC until constant weight. Each capsule with the sample was weighed and placed in an oven (Felisa® St. Livonia, Michigan, U.S.A) for 12 h at 75 ºC. Then, the capsule was removed from the oven and placed in a desiccator. Afterwards, its weight was taken. Moisture content was expressed as a percentage. Fiber content determination Crude fiber was determined using the method proposed by Mexican Norm NMX-F-90-S-1978 (1978). The analysis was performed on the previously defatted sample. First, the sample weight was taken and recorded, then each sample was transferred to a beaker for fiber determination and 200 mL of 1.25% sulfuric acid and 1 mL of isoamyl alcohol as defoamer were added to each beaker. Mixture was kept at boiling point for 30 min. Afterwards, it was rinsed to remove sulfuric acid and isoamyl alcohol residues and at the same time to neutralize the mixture. Subsequently, 200 mL of 1.25% sulfuric acid was added, kept boiling for 30 min and then rinsed in glass fiber until neutralized. Thereafter, fiberglass with the sample in the capsule was placed in an oven (Felisa) and left to dry for 12 h until sample was completely dry. Once the drying process was finished, the capsule with the fiberglass and sample was weighed, and then the fiber percentage of each bean variety was obtained by weight difference. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 5 Carbohydrate content determination Carbohydrates were obtained by difference with ash, fat and moisture content obtained and reported as a percentage. Protein content determination To quantify protein concentration, the Flash 2000 Organic Elemental Analyzer (Thermo Scientific® Corporation, Cambridge, UK) was used, whose procedure is based on the Dumas method proposed by Reussi- Calvo et al. (2008). First, a tin capsule was taken and placed on a microbalance (Mettler Toledo®, Columbus, Ohio, USA), then 9 µg of vanadium pentoxide and 3 µg of the bean sample were weighed. Finally, sample was placed in the Flash 2000 autosampler for analysis. Protein concentration was expressed as a percentage. Energy determination Energy contained in each sample was measured as the sum of calories contained in carbohydrates, fat and protein as described in the Mexican Official Norm NOM-051-SCFI/SSA1-2010 (2010). Energy expressed in Kcal 100-1 g-1. Mineral analysis Micronutrient determination One g of finely ground sample was weighed on an analytical balance (And Company Limited, Milpitas, CA, U.S.A.). Afterwards, acid digestion of the dehydrated plant tissue was performed by adding 25 mL of triacid mixture (1000 mL nitric acid, 100 mL hydrochloric acid and 25 mL sulfuric acid), and placed in a digester (Labconco® Corporation, Kansas City, MO, U.S.A.). After digestion, sample was filtered and volumetrically diluted with tridistilled water in a 50 mL flask. Finally, samples were poured into polypropylene tubes for further analysis. Fe, Zn, Mn, Cu and Ni concentration was determined by atomic absorption spectrophotometry (Atomic Absorption Spectrophotometer iCE 3000 Thermo Scientific® Corporation, Cambridge, UK) and concentration was expressed in ppm. Macronutrient determination Magnesium (Mg), potassium (K), calcium (Ca) were quantified by atomic absorption spectrophotometry (atomic absorption spectrophotometer iCE 3000 Thermo Scientific) in the same way as micronutrients were obtained and expressed as a percentage. Phosphorus (P) was determined by colorimetry using the ammonium metavanadate-molybdate method. 500 µL of the digested sample was taken for minerals, 1 mL of phosphorus reagent (ammonium metavanadate-molybdate) and 3.5 mL of tridistilled water was added; it was stirred in a Vortex (VWR, Thorofare, New Jersey, U.S.A.) and allowed to stand for 1 h. After that time, the samples were read in a UV/Vis spectrophotometer (JENWAY Spectrophotometer, Jenway Limited®, Essex, England) at a wavelength of 430 nm; a calibration curve was performed with a phosphorus standard (Ion Chromatography Standard (IC) AcculonTM Reference Standard, New Haven, Connecticut, U.S.A.) and results were expressed as dry weight percentage (%). C, H, S and N organic compounds determination C, H, S and N were determined by the Dumas method proposed by Reussi-Calvo et al. (2008). First, a tin capsule was taken and placed on a microbalance (Mettler Toledo®, Columbus, Ohio, USA). Nine µg of vanadium pentoxide and 3 µg of the bean sample were weighed, placed in a capsule and then closed. Finally, sample was placed in the Flash 2000 organic elemental analyzer (Thermo Scientific® Corporation, Cambridge, UK) and the compound concentration was reported as a percentage. Three replicates were performed for the analysis. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 6 Antioxidant capacity determination (2, 2-diphenyl-1-picrylhydrazyl (DPPH)) The analysis was carried out using the method proposed by Hsu et al. (2003). Extract was obtained by maceration of 1 g of finely ground seed in 5 mL of 80% methanol, which was centrifuged (Allegra® Refrigerated Centrifuge, Beckman Coulter, Inc.; Fullerton, California, U.S.A.) at 6000 rpm for 10 min at 4 ºC. Once extract was centrifuged, 0.5 mL of the supernatant was taken and mixed with 2.5 mL of freshly prepared 0.1 mM DPPH solution. Thereafter, sample was incubated (Boekel Scientific incubator) for one h in dark conditions and at room temperature. After this time, samples were read in a UV/Vis spectrophotometer (Genesys 10S, Thermo Scientific® Corporation, Cambridge, UK) at a wavelength of 517 nm. Resulting antioxidant capacity was reported as percentage inhibition. Nutraceutical compound determination Total phenols extraction and quantification Phenolic compounds were extracted using the colorimetric method proposed by Singlenton and Rosi (1965). For analysis, 0.5 g of ground bean seed was mixed with 2.5 mL of methanol, 2.5 mL of chloroform, and 1.25 mL of 2% NaCl solution (J. T. Baker, State of Mexico, Mexico) and macerated to obtain an extract. Subsequently, mixture was homogenized, then centrifuged (Allegra® Refrigerated Centrifuge, Beckman Coulter, Inc.; Fullerton, California, U.S.A.) at 5000 rpm for 10 min and three phases were obtained, a methanol phase, which contains the phenolic acids, interphase containing the proteins precipitated by NaCl and the chloroform phase constituted by dissolved lipids. For the reaction, 750 µL of 2% Na2CO3 was placed in a test tube and mixed with 250 µL of Folin-Ciocalteau reagent (Sigma-Aldrich, St. Louis, MO, USA), 1375 µL of deionized water and 250 µL of enzyme extract. Finally, mixture was incubated at room temperature for 60 min. Quantification was obtained using a standard curve of gallic acid (10-100 µg ml-1) at an absorbance of 725 nm. Results are shown in mg of gallic acid equivalents per g of sample (mg GAE g-1) (dry weight). Flavonoid content determination Flavonoid analysis was performed according to the method proposed by Zhishen et al. (1999). Extract was obtained by macerating 0.5 g of ground seed with 5 mL of 85% methanol. Subsequently, it was centrifuged (Allegra® Refrigerated Centrifuge, Beckman Coulter, Inc.; Fullerton, California, U.S.A.) at 4000 rpm for 10 min. Then an aliquot of 250 µL was placed in a test tube, and 75 µL of NaNO2 (J. T. Baker, State of Mexico, Mexico) was added, the mixture was homogenized in a vortex (VWR, Thorofare, New Jersey, U.S.A.) and allowed to stand for 5 min. Afterwards, 150 µL of AlCl3 (Sigma-Aldrich, St. Louis MO, USA) and 500 µL of NaOH (J.T. Baker, State of Mexico, Mexico) were added and diluted to a final volume of 2.025 mL with tridistilled water. Absorbance was then measured at 510 nm in a UV/Vis spectrophotometer (Genesys 10S, Thermo Scientific® Corporation, Cambridge, UK). Results obtained were expressed as mg catechin equivalents (CE) per g of sample (mg CE g-1) based on dry weight. Anthocyanin determination Anthocyanin content was determined by pH differential according to the method proposed by Wrolstad et al. (2005). 0.5 g of finely ground bean was mixed with 5 mL of methanol (J.T. Baker, Estado de Mexico, Mexico). Mixture was centrifuged (Allegra® Refrigerated Centrifuge, Beckman Coulter, Inc.; Fullerton, California, U.S.A) at 4000 rpm for 10 min. After the centrifugation time, 2 phases of the sample were obtained; 0.5 mL of the first phase were taken and placed in a test tube, then 2 mL of potassium chloride (KCl) (J.T. Baker, Estado de México, México) were added, homogenized in a vortex (VWR, Thorofare, New Jersey, U.S.A.) and its absorbance was obtained in a UV/Vis spectrophotometer (Genesys 10S, Thermo Scientific® Corporation, Cambridge, UK) at 460 nm. Subsequently, 0.5 mL of the second phase was taken and deposited in a test tube, 2 mL of sodium acetate was added, homogenized in the vortex and the absorbance Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 7 reading was taken in the UV/Vis spectrophotometer. Results were expressed as mg Cyanidin-3-glucoside (C3G) g-1 (dry weight)-1. Phytic acid determination For phytic acid determination, the method proposed by McKie and McCleary (2016) was used, first total phosphorus was obtained by the ammonium metavanadate-molybdate method in an absorption range of 430 nm and a potassium phosphate standard curve. Once the results were obtained, the following formula was applied: Phytic acid = (Total phosphate) /0.282 (5) Where 0.282 is a conversion factor from total phosphorus to phytic acid. Results were expressed in g of phytic acid per 100 g (g 100-1 g-1) based on dry weight of sample. Statistical analysis Data was subjected to an analysis of variance in a completely randomized one-factor design to evaluate the effect of bean variety on the different variables studied, as well as an analysis of comparison of means using the Tukey test and an analysis of correlation between variables performed with SAS statistical package (SAS Institute, INC; Cary, NC, USA) Means were accepted as significantly different at a 95% confidence interval (p ≤ 0.05). Results were reported as mean± standard deviation. Results Results Results Results and Discussionand Discussionand Discussionand Discussion Morphometric characteristics of bean seeds Table 2 shows weight, length, width and thickness of bean seeds of the evaluated varieties. Statistical analysis showed a significant difference (p ≤ 0.05) between varieties for the four variables mentioned above. Seed width ranged from 5.5 to 9.1 mm, with the lowest value for ‘Flor de Mayo’ and the highest for ‘Sangre de Toro’, the latter being the only variety with a statistical difference compared to other varieties. Bean thickness ranged from 4.9 to 6.8 mm, with ‘Pinto’ being the lowest and ‘Peruano’ the highest, although the latter variety was statistically equal to ‘Flor de Mayo’, ‘Negro’/102 and ‘Negro’/104. In addition, kernel length ranged from 10.8 to 17.3 mm, with ‘Negro’/102 being the shortest variety and ‘Sangre de Toro’ the longest. Finally, the weight of 100 seeds ranged from 23.06 to 53.42 g, with ‘Negro’/102 being the shortest and ‘Sangre de Toro’ the heaviest. In summary, ‘Sangre de Toro’ had the highest values for width, length and weight among all analysed varieties, while ‘Negro’/102 was the shortest and lightest. Results obtained from dimensions of the bean varieties analysed in the present work are lower than those reported by Herrera-Hernández et al. (2018) in bean varieties grown in Zacatecas, Mexico. Likewise, the weight of 100 seeds was higher than reported by Mederos and Reynaldo (2007) in ‘Cuban bean’ varieties with black and red coat. According to the classification of Aguirre and Gómez-Aldapa (2010), ‘Sangre de Toro’ and ‘Peruano’ varieties are classified as large (>40 g) and ‘Flor de Mayo’, ‘Negro’/102, ‘Pinto’ and ‘Negro’/104 as medium (26 to 40 g). Other studies have shown great variability in the weight of 100 seeds, for example Pliego- Marín et al. (2013) found a large amplitude of this variable in seeds collected in Central Valleys of Oaxaca, Mexico, with intervals ranging from 11.2 to 74.8 g of 100 seeds, being out of the range of what was found in this study, with very small genotypes such as ‘Negro Delgado’ with a weight of 11.2 g of 100 seeds coming from Zaachila and ‘Frijolon’ with 74.8 g weight with the same origin. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 8 Table 2.Table 2.Table 2.Table 2. Morphometric characteristics of several common bean varieties from Morelos State, Mexico Variety/ Classification number* Width (mm) Thickness (mm) Length (mm) Weight (g of 100 seeds) ‘Sangre de Toro’/98 9.1 ± 0.01a 6.4 ± 0.02ab 17.3 ± 0.09a 53.42 ± 3.59a ‘Peruano’/100 8.2 ± 0.05b 6.8 ± 0.06a 13.6 ± 0.04b 44.41 ± 1.01b ‘Flor de Mayo’/101 5.5 ± 0.02b 5.5 ± 0.02bc 12.6 ± 0.05bc 30.97 ± 0.18c ‘Negro’/102 7.3 ± 0.02 b 5.5 ± 0.02bc 10.8 ± 0.03d 23.06 ± 0.33d ‘Pinto’/103 7.5 ± 0.07b 4.9 ± 0.03c 13.1 ± 0.09bc 29.07 ± 0.36c ‘Negro’/104 7.4 ± 0.02b 5.6 ± 0.05bc 11.46 ± 0.04cd 30.29 ± 0.84c Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey test (p ≤ 0.05). *Laboratory work classification. Bean seed color characteristics Bean color is an important aspect for consumer taste so its evaluation is necessary (Aguirre-Santos et al., 2011). Table 3 presents the colour characteristics L*, a*, b*, chroma and hue of common bean seeds from Morelos, Mexico, the statistical analysis showed significant difference (p ≤ 0.05) between varieties in all five variables analysed. Luminosity ranged from 24.96 to 69.21, with ‘Negro’/102, ‘Negro’/104 and ‘Sangre de Toro’ being the bean varieties with the lowest luminosity corresponding to darker bean colors. While ‘Peruano bean’, ‘Pinto’ and ‘Flor de Mayo’ had the highest L* value corresponding to lighter colors of all varieties analysed. These values correspond to those reported by Chávez-Mendoza et al. (2019) in bean varieties from different regions of Mexico. They are also similar to those reported by Aguirre and Gómez-Aldapa (2010) for ‘Pinto Saltillo’, ‘Bayo Victoria’ and ‘Negro San Luis’ bean varieties with 7.125, 57.1 and 21.85, respectively. Regarding the a* value, it ranged from 0.71 to 26.043. Where ‘Peruano’ and ‘Negro’/104 beans had the lowest values for this color characteristic, with no statistical difference between them, indicating a lower tendency to red and a higher tendency to green. In addition, ‘Sangre de Toro’ had the highest a* value of all varieties analysed, which coincides with its strong red color. On the other hand, b* value was within a range of -1.42 to 33.44. ‘Negro’/102 and ‘Negro’/104 had negative values indicating a greater tendency to blue color, while ‘Peruano’ bean had the highest b* value with a more yellow tendency. Other studies reported negative values of b* for dark-skinned beans such as ‘Negro Puebla’, ‘Negro 151’, ‘Negro 152’, ‘Negro Querétaro’, ‘Negro San Luis’, ‘Negro Sinaloa’, ‘Negro Veracruz’, ‘Medellín’, ‘Nayarit 80’, ‘Jamapa’, ‘Negro Perla’, ‘Merentral’, ‘Altiplano’ and ‘Negro Puebla 152’, which places them in the third quadrant of the tri-stimulus hunter L scale, a* b* corresponding to blue-green coloration (Salinas-Moreno et al., 2005). Regarding chromaticity, it showed a range from 1.77 to 33.45, with ‘Negro’/102 and ‘Negro’/104 having the lowest values or lowest color saturation, whereas the ‘Peruano bean’ had the highest color clarity or chromaticity while ‘Flor de Mayo’ and ‘Negro’/104 were in the intermediate range for this variable. This range was higher than that reported by Aguirre and Gómez-Aldapa (2010) in ‘Negro San Luis’, ‘Pinto Saltillo’ and ‘Bayo Victoria’ varieties. Hue angle was within the range of 11.75 ° and 126.64 °, with ‘Sangre de Toro’ variety having the lowest value, which is coincidentally in the red tone of the color wheel (McGuire, 1992), while ‘Negro’/102 variety had the highest angle with a bluish-green tone, equal to the coloration reported by Aguirre and Gómez-Aldapa (2010) for ‘Negro San Luis’ bean, which corresponds to the third quadrant of the tristimulus scale. Likewise, ‘Peruano’ and ‘Pinto’ varieties showed a shade closer to yellow, since their hue angle was close to 90 °, as reported by McGuire (1992). Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 9 Table 3.Table 3.Table 3.Table 3. Seed color characteristics of several common bean varieties from Morelos State, Mexico Variety/ Classification number* L a* b* Chroma ° Hue ‘Sangre de Toro’/98 32.41 ± 0.09d 26.04 ± 1.03a 5.42 ± 0.20d 26.60 ± 1.04b 11.75 ± 0.25f ‘Peruano’/100 69.21 ± 1.1a 0.71 ± 0.68d 33.44 ± 1.07a 33.45 ± 1.07a 88.78 ± 1.75c ‘Flor de Mayo’/101 50.14 ± 0.71c 14.81 ± 0.51b 11.46 ± 0.59c 18.73 ± 0.58c 37.74 ± 1.62e ‘Negro’/102 24.96 ± 0.40f 1.05 ± 0.11d -1.42 ± 0.09e 1.77 ± 0.05d 126.64 ± 4.49a ‘Pinto’/103 65.94 ± 0.67b 6.21 ± 0.24c 16.46 ± 0.06b 17.59 ± 0.05c 69.32 ± 0.08d ‘Negro’/104 26.85 ± 0.60e 0.91 ± 0.07d -1.80 ± 0.12e 2.03 ± 0.07d 116.99 ± 3.55b Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey (p ≤ 0.05). *Laboratory work classification. Nutritional analysis Table 4 reports the nutritional composition of the bean varieties analysed in the present study. Statistical analysis showed significant differences (p ≤ 0.05) between varieties in protein, ash, fat, moisture, carbohydrate, fiber and energy content. Protein concentration ranged from 18.03 to 26.92%. ‘Sangre de Toro’ and ‘Pinto’ varieties had the lowest content, ‘Flor de Mayo’, ‘Negro’/102 and ‘Negro’/104 had intermediate values and ‘Peruano’ bean showed the highest concentration. The results obtained in most of the varieties analysed in this study are similar to those reported by Armendáriz-Fernández et al. (2019) in bean varieties harvested in Oaxaca, Mexico. As well as to those found by Herrera-Hernández et al. (2018) in varieties harvested in Zacatecas, Mexico. They also coincide with the results obtained by Peña-Betancourt and Conde-Martínez (2012) on wild bean varieties (‘Durango Atypical’ and ‘Typical’, ‘Oaxaca Chico’ and ‘Tlaxcala Atypical’ and ‘Typical’). Also, commercial beans such as ‘Flor de Mayo’, ‘Peruano’, ‘Garbancillo’ and ‘Flor de Junio’. Mederos (2006), on the other hand, indicates that protein in beans ranges between 16 and 30% and that varieties most consumed in Latin America have an average concentration of 20%, which is in agreement with results obtained in the present study. According to the same author, bean protein has a high lysine and phenylalanine plus tyrosine content, so it fulfils all the minimum requirements recommended by the Food and Agriculture Organization (FAO) or the World Health Organization (WHO). Ash content ranged from 4.0 to 4.96%. ‘Flor de Mayo’ and ‘Negro’/104 varieties had the lowest concentration with no statistical difference between them, followed by ‘Pinto’, ‘Negro’/102, and ‘Peruano’ with intermediate values, and finally ‘Sangre de Toro’ with the highest concentration among all six varieties analysed. The obtained range was higher than reported by Herrera-Hernández et al. (2018) in varieties harvested in Zacatecas, Mexico and slightly lower than results obtained by Armendariz-Fernández et al. (2019) in bean varieties harvested in the state of Oaxaca, Mexico. Aguirre and Gómez-Aldapa (2010) reported slightly lower results in ‘Negro San Luis’, ‘Pinto Saltillo’ and ‘Bayo Victoria’ varieties, noting that ash content may vary depending on cultivar genetics and soil characteristics. Fat concentration was within the range of 0.96 to 1.64%. ‘Negro’/104 had the lowest value for this variable while ‘Peruano’ had the highest content of all evaluated variables, with no statistical difference between ‘Sangre de Toro’ and ‘Negro’/102. These results are similar to those reported by Aguirre and Gómez-Aldapa (2010) on ‘Negro San Luis’, ‘Pinto Saltillo’ and ‘Bayo Victoria’ varieties corresponding to a range of 0.92 to 1.71%. In addition, they are higher than those reported by Fernandez and Sanchez (2017). Lipid fraction of bean is the smallest, and is constituted by a mixture of acylglycerides whose predominant fatty acids are monounsaturated and polyunsaturated (Ulloa et al., 2011). Moisture content of common bean was within the range of 10.29 and 14.58%, being ‘Negro’/104 the variety with the lowest moisture content and ‘Negro’/102 with the highest value. Results obtained in this last variety were much higher than those reported in other black beans such as ‘Negro San Luis’, which had a Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 10 moisture content of 11.95 % (Aguirre and Gómez-Aldapa, 2010), but were similar to those found in some Cuban varieties (Mederos and Reynaldo, 2007); however, the remaining varieties analysed in the present work were much lower than the values reported by the latter authors. Other studies have shown results different from those found in this work such as the one conducted by Armendariz-Fernández et al. (2019) who reported that ‘Sangre de Toro’ and ‘Peruano’ varieties had a lower moisture content, while the ‘Flor de Mayo’ variety had a higher amount (12.8%) than that observed in this study. Meanwhile, Herrera-Hernández et al. (2018) reported much lower moisture values in bean varieties produced in Zacatecas, Mexico, ranging from 6.14 to 7.42%. Peña-Betancourt and Conde-Martínez (2012) found much lower results in wild and improved bean varieties, suggesting that the lower moisture content of wild varieties was due to increased temperature and lack of irrigation at the production site, in addition to prolonged storage. Aguirre and Gómez-Aldapa (2010) reported that moisture content is related to seed age and postharvest handling, as well as to processing methods and conditions. Carbohydrates constitute the main fraction in legume beans (Mederos, 2006), 100 g of raw beans provide 52 to 76 g depending on variety (Ulloa et al., 2011). Carbohydrate concentration obtained in the present study was between 51.21 and 61.53%, where ‘Pinto’ variety had the highest content, while ‘Peruano’ had the lowest. No statistical difference was obtained between the concentration of ‘Negro’/104 (58.28%) and ‘Sangre de Toro’ (58.01%), nor between ‘Flor de Mayo’ (53.58%) and ‘Negro’/102 (53.67%), which had intermediate values for this variable. Other studies have shown values different from these results; thus, Armendariz-Fernández et al., (2019), reported a higher carbohydrate content in ‘Flor de Mayo’ (55 %) and ‘Peruano’ (56.6%) varieties, while in ‘Sangre de Toro’ beans the found value was lower (55.2%). For their part, Herrera-Hernández et al. (2018) reported a higher range (57.16% to 65.79%) of carbohydrate content in common bean varieties grown in Zacatecas, Mexico, including ‘Flor de Mayo’, ‘Negro’, ‘Pinto Saltillo’ bean, among others. While Fernandez and Sanchez (2017) found much lower values than those obtained in the present study in different varieties of beans produced and consumed in Mexico purchased in a local market in Delicias, Chihuahua, Mexico, such as ‘Bayo’, ‘Pinto’, ‘Negro’, ‘Alubia’, ‘Flor de Mayo’ and ‘Peruano’, where the latter was the exception as it showed a higher concentration than that obtained in this study. Bean carbohydrates consist mainly of starch and other polysaccharides (dietary fiber) with small but significant amounts of oligosaccharides; starch represents more than 50% of seed weight and is the dominant carbohydrate in the human diet, hence the importance of this legume (Mederos, 2006). Bean is also a good source of fiber which ranges in value from 14-19 g 100-1 g-1 of the raw food, from which up to half may be of the soluble form. The main chemical components of fiber in beans are pectins, pentosans, hemicellulose, cellulose and lignin (Ulloa et al., 2011). Half a cup of beans provides between 5.2 and 7.8 g of total fiber (Messina, 2014). Fiber content in the present study ranged from 3.21 to 5.71%, ‘Flor de Mayo’ variety had the highest value while ‘Negro’/104 bean had the lowest. These results were far higher than those reported by Herrera-Hernández et al. (2018) in genotypes grown in Zacatecas, Mexico and to those reported by Armendariz-Fernández et al. (2019) on varieties produced in Oaxaca, Mexico. Finally, the energy content of the analysed beans was between 318.39 and 334.6 Kcal, with ‘Negro’/104 being the variety with the highest energy content and ‘Negro’/102 the lowest. Similar results were reported by Armendariz-Fernández et al. (2019) on several bean varieties produced in Oaxaca, Mexico, including ‘Sangre de Toro’, ‘Bayo’, ‘Peruano’ among others. Likewise, results were lower than those reported by Herrera- Hernández et al. (2018) in bean varieties produced in Zacatecas, Mexico. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 11 Table 4.Table 4.Table 4.Table 4. Nutritional composition of several common bean varieties from Morelos State, Mexico Variety/ Classification number* Protein (%) Ash (%) Fat (%) Moisture (%) Carbohydrates (%) Crude fiber (%) Energy (Kcal 100-1g-1) ‘Sangre de Toro’/98 18.03 ± 0.13d 4.96 ± 0.03a 1.53 ± 0.04a 13.45 ± 0.04b 58.01 ± 0.15b 3.40 ± 0.02c 320.41 ± 0.23d ‘Peruano’/10 0 26.92 ± 1.87a 4.22 ± 0.02c 1.64 ± 0.05a 11.93 ± 0.02c 51.21 ± 0.10d 3.81 ± 0.02b 328.36 ± 0.13c ‘Flor de Mayo’/101 23.29 ± 1.40b 4.0 ± 0.02d 1.11 ± 0.07cd 11.53 ± 0.02d 53.58 ± 0.12c 5.71 ± 0.02a 320.59 ± 0.39d ‘Negro’/102 21.72 ± 1.15bc 4.26 ± 0.01c 1.47 ± 0.07ab 14.58 ± 0.03a 53.67 ± 0.12c 3.4 ± 0.02c 318.39 ± 0.55e ‘Pinto’/103 19.66 ± 1.33cd 4.45 ± 0.05 b 1.26 ± 0.13bc 10.63 ± 0.02e 61.53 ± 0.15a 3.31 ± 0.04d 332.74 ± 0.80b ‘Negro’/104 22.71 ± 0.66bc 4.05 ± 0.05d 0.96 ± 0.05d 10.29 ± 0.01f 58.28 ± 0.13b 3.21 ± 0.03e 334.6 ± 0.14a Data correspond to mean ± SD. Means with the same letter between rows indicate that there is no statistical difference between varieties. Tukey test (P ≤ 0.05). *Laboratory work classification. Mineral content analysis Micronutrient content Table 5 shows the micronutrients present in the common bean varieties produced in Morelos State, Mexico. Statistical analysis showed a significant difference (p < 0.05) between varieties in the content of all the microelements analyzed. Iron (Fe) concentration ranged from 89.225 to 136.416 ppm, with ‘Flor de Mayo’ and ‘Sangre de Toro’ varieties having the lowest Fe content and no significant difference between them, while ‘Pinto beans’ had the highest concentration. These results are higher than those reported by Akond et al. (2011a) on 14 genotypes from the International Center for Tropical Agriculture and the United States of America. As well as those obtained by Armendáriz-Fernández et al. (2019) on bean varieties produced and consumed in Oaxaca, Mexico. On the other hand, they are lower than those obtained in most of the bean varieties analyzed by Chávez- Mendoza et al. (2019), who found that this micronutrient is present in greater proportion in the coat than in the bean cotyledon. Fe deficiency has been mainly related to anemia. This micronutrient is also part of a large number of enzymes involved in energy production and in the proper functioning of the immune response in humans. Beans contribute approximately 40% of Fe to the diet of people who base their diet on this legume and corn (Mederos, 2006), hence the importance of studying this micronutrient in the different genotypes produced in Mexico, which serves as a basis for biofortification and genetic improvement studies. Zinc (Zn) content ranged from 24.82 to 35.78 ppm, ‘Flor de Mayo’ variety had the lowest value, while ‘Peruano’ had the highest. These results were similar to those reported by Armendáriz-Fernández et al. (2019) in bean varieties produced and consumed in the state of Oaxaca Mexico. Some studies report that Zn content in beans is one of the highest among vegetables, almost equal to that found in dairy products, although lower than that found in meat. Evaluations carried out on bean collections reveal ranges in the content of this micronutrient from 21 to 54 ppm with an average of 35 ppm (Mederos, 2006), which coincides with the results found in this work. Importantly, this microelement is associated with decreased oxidative stress in cells and improved immune cell function. In addition, its deficiencies have been shown to cause DNA damage in peripheral blood cells in rats (Mudryj et al., 2014). Nickel (Ni) concentration in analyzed varieties ranged from 2,113 to 6,783 ppm with ‘Negro’/102 bean having the lowest value and ‘Sangre de Toro’ having the highest. These data are similar to those reported by Chávez-Mendoza et al. (2019) in different bean varieties, where a higher presence of this micronutrient was observed in the cotyledon than in the coat, with ‘Bayo’ having the highest value at 8.62 ppm, while ‘Negro’ and ‘Flor de Mayo’ beans had the lowest, coinciding with what was reported in the present study. Manganese (Mn) content ranged from 9.036 to 18.82 ppm, with ‘Negro’/102 having the lowest concentration of this micronutrient, while ‘Pinto’ had the highest value. These results were similar to those reported by Chávez-Mendoza et al. (2019) in several bean varieties, including ‘Flor de Mayo’, ‘Pinto Saltillo’, Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 12 ‘Negro San Luis’, ‘Negro 8025’, ‘Negro Jamapa’, ‘Higuera Azufrado’ among others. However, they are lower than those obtained by Herrera-Hernández et al. (2018) in some varieties grown and consumed in Zacatecas, Mexico, including the ‘Flor de Mayo’ variety in which a higher content of this microelement was obtained (25.45 pm) than that obtained in the present work. Lastly, Cu concentrations of the varieties studied ranged from 3.48 to 6.69 ppm. ‘Sangre de Toro’ had the lowest content of this micronutrient while ‘Pinto’ had the highest. These results are much lower than those recorded by Herrera-Hernández et al. (2018) in bean varieties grown in Zacatecas state, Mexico. Summarizing, ‘Pinto’ had the highest concentration of Fe and Mn, ‘Peruano’ had the highest concentration of Cu and Zn, while ‘Sangre de Toro’ had the highest Ni content and the lowest amount of Fe and Cu. The average concentration of these micronutrients in the analysed seed in descending order was as follows: Fe > Mn > Zn> Cu > Ni. This behaviour is similar to that reported by Chávez-Mendoza et al. (2019) in several bean varieties, with the exception of manganese, which, unlike the present study, was in lower concentration than zinc. Observed differences in mineral content among the varieties analysed in this study can be attributed to the genotype and the environment in which they are produced. Previous studies have shown that the concentration of these compounds in beans has varied as a function of genetic material, crop management and storage conditions (Espinoza-García et al., 2016). Table 5.Table 5.Table 5.Table 5. Micronutrient concentration in common bean varieties produced in Morelos State, Mexico Variety/ Classification number* Fe (ppm) Zn (ppm) Ni (ppm) Mn (ppm) Cu (ppm) ‘Sangre de Toro’/ 98 89.22 ± 1.65c 32.97 ± 3.70ab 6.78 ± 0.21a 13.27 ± 0.64bc 3.48 ± 0.42b ‘Peruano’/ 100 101.40 ± 3.24bc 35.78 ± 5.10a 3.28 ± 0.46c 11.05 ± 0.41cd 6.69 ± 0.50a ‘Flor de Mayo’/ 101 91.06 ± 6.49c 24.82 ± 0.66b 2.45 ± 0.33cd 10.54 ± 0.65cd 3.95 ± 0.68b ‘Negro’/ 102 99.76 ± 5.83bc 31.85 ± 1.133ab 2.11 ± 0.30d 9.036 ± 1.43d 6.09 ± 0.56a ‘Pinto’/ 103 136.41 ± 10.01a 31.81 ± 3.48ab 5.23 ± 0.46b 18.82 ± 1.83a 4.21 ± 0.68b ‘Negro’/ 104 113.43 ± 6.09b 34.38 ± 0.90a 2.49 ± 0.28cd 14.491 ± 0.73b 4.39 ± 0.38b Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey (p ≤ 0.05). *Laboratory work classification. Macronutrient determination Ca, Mg and K are the main cations in common bean. There is greater availability of Ca than Mg or K (Suárez-Martínez et al., 2016). Table 6 shows the P, K, Mg and Ca contents obtained in the bean samples of the different varieties analysed. Statistical analysis showed a significant difference (p < 0.05) between varieties in Mg concentration but not in P, K and Ca content. Phosphorus, plays a relevant role in various metabolic processes vital to all living organisms such as being part of macromolecular structures, in energy generation and metabolic regulation, hence the importance of its availability in free form as inorganic P, which can be accessed by organisms' cells, as well as other nutrients that are chelated (Rodriguez-Blanco et al., 2018). P concentration in bean genotypes evaluated was within a range of 0.149 to 0.194%, although no statistical difference was observed between varieties for this variable, ‘Negro’/104 bean had the lowest concentration of this macronutrient while ‘Negro’/102 had the highest. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 13 Results obtained in the present study were higher than those described by Armendáriz-Fernández et al. (2019) in common beans produced in Oaxaca, Mexico, where ‘Peruano’ and ‘Sangre de Toro’ had the highest concentrations with 0.17% for both, and varieties such as ‘Negro Michigan’ and ‘Bayo Bola’ had concentrations as low as 0.01%. Fernandez and Sanchez (2017) meanwhile reported higher P values than those obtained here in ‘Pinto’, ‘Peruano’ and kidney bean. As for K concentration, it ranged from 1.464 to 1.815%. These results are higher than those recorded by Armendáriz-Fernández et al. (2019) in common bean produced in the State of Oaxaca, Mexico. They also exceed the concentrations of this macronutrient obtained by Fernandez and Sanchez (2017) in beans commonly consumed in Mexico such as ‘Bayo’, ‘Pinto’, ‘Peruano’, ‘Negro’, kidney bean, ‘Flor de Mayo’ and green bean. Ca, Mg and Cu are deficient in developed and developing countries which is attributable to the low availability of mineral elements in the soil and/or the low ability of plants to store them in their tissues, associated at the same time with the scarce availability of food of animal origin (Araméndiz-Tatis et al., 2016). For the present study, the Mg content in the analysed beans ranged from 0.169 to 0.201%, with the ‘Negro’/104 bean having the lowest concentration, while the ‘Negro’/102 bean had the highest amount. Results were much higher than those reported by Herrera-Hernández et al. (2018) in common bean varieties produced in Zacatecas, Mexico, in which ‘Patola’ and ‘Japanese’ beans had the highest concentrations of this macronutrient with a concentration of 0.16% in both genotypes. Fernandez and Sanchez (2017) also found lower concentrations than those obtained here, in bean varieties produced and consumed in Mexico. Ca belongs to the group of minerals that should always be part of our diet. It is the most abundant mineral element in our body, as it is an important part of the skeleton and teeth. It accounts for about 2% of body weight and is an essential cellular component for maintaining and/or performing the various specialized functions of virtually all cells in the body. These functions, non-skeletal, we can divide them into structural and properly regulatory (Martinez, 2016). In this study, Ca content ranged from 0.154 to 0.273%. These results are similar to those covered by Herrera-Hernández et al. (2018) on common bean varieties produced in Zacatecas, Mexico. Other studies have disclosed lower concentrations than those obtained in the present investigation in ‘Pinto’, ‘Flor de Mayo’ and ‘Peruano’ beans (Fernandez and Sanchez, 2017). In summary, ‘Negro’/102 had the highest Mg concentration while ‘Negro’/104 had the lowest. Moreover, all the varieties analysed had the same P, K and Ca contents according to the statistical analysis. Table 6.Table 6.Table 6.Table 6. Macronutrient concentration (%) in common bean varieties produced in Morelos State, Mexico Variety/ Classification number* P K Mg Ca ‘Sangre de Toro’/ 98 0.180 ± 0.009a 1.815 ± 0.04a 0.192 ± 0.002ab 0.228 ± 0.073a ‘Peruano’/ 100 0.182 ± 0.026a 1.655 ± 0.048a 0.188 ± 0.006bc 0.1545 ± 0.022a ‘Flor de Mayo’/ 101 0.170±0.015a 1.659 ± 0.033a 0.173 ± 0.003d 0.218 ± 0.0651a ‘Negro’/ 102 0.194 ± 0.084a 1.605 ± 0.315a 0.201 ± 0.002a 0.273 ± 0.025a ‘Pinto’/ 103 0.175 ± 0.059a 1.464 ± 0.080a 0.179 ± 0.001dc 0.245 ± 0.034a ‘Negro’/ 104 0.149 ± 0.012a 1.687 ± 0.062a 0.169 ± 0.001d 0.239 ± 0.058a Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey test (p ≤ 0.05). *Laboratory work classification. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 14 Organic compound determination C, S, N, H Table 7 presents the C, S, N and H contents obtained in the common bean samples of the different varieties analysed. Statistical analysis showed significant difference (p < 0.05) between varieties in N concentration, but not in C, S, and H content (p > 0.05). Carbon concentration in the analysed samples ranged from 40.441 to 42.349%, S from 0.040 to 0.082% and H from 6.153 to 6.391% with no significant statistical difference between varieties for the three elements. These results are lower than those shown by Chávez-Mendoza et al. (2019) in several bean varieties, in which no significant difference was observed between varieties in the cotyledon, as occurred in the present investigation; however, these authors did observe significant differences in the content of these elements in the seed coat. On the other hand, Paredes et al. (2009) found C and S concentrations higher than those found in the present study in Chilean, Nuevo Granada, Durango and Mesoamerican breeds. Nitrogen content ranged from 2.926 to 4.0%, with ‘Sangre de Toro’ variety having the lowest concentration and ‘Peruano’ the highest. These results are lower than those obtained by Chávez-Mendoza et al. (2019) in several bean varieties, both in coat and cotyledon, including ‘Flor de Mayo’, ‘Pinto Saltillo’, ‘Negro Jamapa’, ‘Negro San Luis’, ‘Negro 8025’, among others. Summarizing, it was observed that ‘Sangre de Toro’ variety presented the lowest values in N concentration, while ‘Peruano’ bean and ‘Flor de Mayo’ had the highest amount of this compound. Furthermore, all varieties analysed had statistically the same C, S and H concentration. Table 7.Table 7.Table 7.Table 7. C, S and N concentration (%) in common bean varieties produced in Morelos State, Mexico Variety/ Classification number* C S N H ‘Sangre de Toro’/98 40.441 ± 0.53a 0.087 ± 0.018a 2.926 ± 0.12c 6.153 ± 0.203a ‘Peruano’/100 41.150 ± 0.11a 0.097 ± 0.010a 4.000 ± 0.21a 6.332 ± 0.064a ‘Flor de Mayo’/101 41.418 ± 0.69a 0.094 ± 0.023a 3.660 ± 0.28ab 6.315 ± 0.166a ‘Negro’/102 41.625 ± 1.00a 0.086 ± 0.019a 3.542 ± 0.232abc 6.391 ± 0.176a ‘Pinto’/103 42.349 ± 2.74a 0.040 ± 0.036a 3.068 ± 0.43bc 6.513 ± 0.49a ‘Negro’/104 41.598 ± 0.52a 0.082 ± 0.015a 3.542 ± 0.13abc 6.347 ± 0.098a Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey test (p ≤ 0.05). *Laboratory work classification. Antioxidant capacity and nutraceutical compounds Antioxidant capacity The antioxidant capacity obtained in the analysed bean varieties is presented in Figure 1. Statistical analysis showed significant difference (p ≤ 0.05) between varieties in this variable, which ranged from 73.59 to 96.76% inhibition. ‘Peruano’ bean had the lowest antioxidant capacity, with no statistical difference with ‘Pinto’ and ‘Negro’/104, while the highest value was for the ‘Flor de Mayo’ variety, which was statistically equal to ‘Sangre de Toro’ and ‘Negro’/102. Study by Chávez-Mendoza et al. (2019) also revealed ‘Flor de Mayo’, ‘Negro 8025’ and ‘Negro San Luis’ as the varieties with the highest antioxidant capacity, finding differences in this variable between cotyledon and coat, with the latter having the highest percentage of inhibition. These authors noted that these varieties had the highest phenolic content, which could explain their higher antioxidant activity, as was the case in the present study, where ‘Flor de Mayo’, ‘Sangre de Toro’ and ‘Negro’/102 had the highest concentration of total phenols, flavonoids and anthocyanins. Silva-Cristobal et al. (2010) also disclosed that the antioxidant capacity of legumes depends on the total polyphenol content. Aguilera et al. (2011) point out that the antioxidant activity is directly related to the polyphenol structure, such as the number of hydroxyl groups, degree of glycosylation etc. González de Mejía et al. (1999) found that phenolic compounds extracted from the coat of common bean inhibit the mutagenicity induced by Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 15 benzopyrene and 1-nitropyrene (premutagenic and mutagenic agents, respectively), hence the importance of studying these compounds present in common bean. On the other hand, Armendariz-Fernández et al. (2018) reported lower results in bean varieties grown in Oaxaca, Mexico also finding that ‘Sangre de Toro’ and ‘Flor de Mayo’ variety along with two other varieties, had the highest values of antioxidant ability with 82.1% and 79.1% respectively. Some authors have stated that ‘Flor de Mayo’ is the preferred variety for consumption in Central Mexico (Cardador-Martínez et al., 2002), which confirms the importance of this variety for Mexican consumers. Figure 1Figure 1Figure 1Figure 1. Antioxidant capacity of common bean varieties grown in Morelos State, Mexico. Means with the same letter are not significantly different, Tukey test (p ≤ 0.05). *Laboratory work classification. Nutraceutical compounds Table 8 shows the nutraceutical compounds evaluated in common bean varieties produced in Morelos State, Mexico. Statistical analysis showed significant difference (p ≤ 0.05) between varieties in the content of total phenols, flavonoids and anthocyanins, but not in the phytic acid concentration (p > 0.05). Total phenol content ranged from 1.76 to 4.24 (mg GAE g-1). Statistical analysis showed significant differences between varieties in the content of this compound. ‘Negro’/104 was the variety with the lowest phenol concentration along with ‘Negro’/102, whereas ‘Flor de Mayo’ had the highest content, followed by ‘Sangre de Toro’, while ‘Pinto’ and ‘Peruano’ beans had intermediate concentrations. Other studies have found that these compounds are found in greater amounts in the bean coat than in the cotyledon; and have reported results similar to those obtained in the present work in some common Mexican bean varieties, such as ‘Flor de Mayo’, ‘Negro 8025’, ‘Negro San Luis’, ‘Pinto Saltillo’ among others (Chávez-Mendoza et al., 2019). Likewise, the results obtained were superior to those found by Espinosa-Alonso et al. (2006) in wild and weedy Mexican bean germplasm materials from Chiapas, Chihuahua, Durango, Guerrero and Jalisco, as well as in ‘Pinto’ and ‘Negro Jamapa’ beans. The total phenol level obtained in the bean of the present study exceeds that observed in wild berry species of the genus Vaccinium, which are among the most important sources of these bioactive compounds in fruits with a content of 0.81 to 1.70 mg GAE g-1 (Taruscio et al. 2004), which denotes the importance of consumption of the common bean studied in this work. According to Rodriguez et al. (2021) the differences in phenolic composition between varieties may be related to the color of the seed coat, however, other studies have suggested that this variability is due more to genotype than to color, as well as to environmental conditions under which it is produced. a b a a ab ab 0 20 40 60 80 100 120 ´Sangre de Toro´/98 ´Peruano´/100 ´Flor de Mayo´/101 ´Negro´/102 ´Pinto´/103 ´Negro´/104 A n ti ox id an t C ap ac it y of c om m on b ea n v ar ie ti es (% i n h ib it io n ) Bean variety/clasification number Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 16 Phytic acid is bound to minerals and does not allow mineral availability due to its chelating property, it has been reported to inhibit the absorption of Fe, Zn, Ca, Mg and Mn (Gupta et al., 2015). However, it is also considered an antioxidant and anti-carcinogen with potential human health benefits. This accounts for 60 to 85% of the total P in the seed (Akond et al., 2011b). In the present study, phytic acid concentration ranged from 0.149 to 0.194 g 100-1 g-1, with no significant difference between varieties. Similar results were disclosed by Iniestra-González et al. (2005) on a group of 16 Mexican common bean varieties with different bean types, ‘Negro’, ‘Pinto’, ‘Crema’ or ‘Bayo’, ‘Azufrado’, ‘Flor de Mayo’ and ‘Blanco’. While they were much lower than those obtained by Díaz-Batalla et al. (2006) in wild and cultivated varieties of Mexican common raw and cooked beans and by De Paula et al. (2018) in different genotypes of Colombian cowpea, in which the control ‘Criollo Córdova’ showed the maximum value with 12.27 mg g-1. On the other hand, the results obtained were superior to those shown by Akond et al. (2011a) in several common bean genotypes from USA, Brazil and the International Center for Tropical Agriculture. Although phytic acid is considered an anti-nutrient, it does not represent a problem and can positively affect human health if a diverse diet is consumed in which micronutrient intake and bioavailability are high (Diaz-Batalla et al., 2006). Flavonols quercetin and kaemferol are the most important flavonoids in foods and their consumption has been linked to an inverse association between lung cancer and risk of cardiovascular disease (Díaz-Batalla et al., 2006). Their presence affects the flavour and color of common bean (Yang et al., 2018). In the present research, flavonoids were present in a concentration ranging from 0.275 to 1.991 mg CE g-1. ‘Peruano’ bean had the lowest concentration, which showed no significant difference with ‘Negro’/104, ‘Negro’/102 and ‘Pinto’ bean. Whereas ‘Flor de Mayo’ had the highest content of these nutraceutical compounds, whose concentration was statistically equal to that obtained in the ‘Sangre de Toro’ variety. Result obtained in ‘Flor de Mayo’ was slightly higher than that reported by Herrera-Hernández et al. (2018) in this same variety but grown in Zacatecas, Mexico, in general the analysed varieties in this study had a higher content of flavonoids than those evaluated by those authors such as ‘Bayo’ bean, ‘Flor de Junio’, ‘Reata’, ‘Canario’, ‘Pinto Saltillo’, ‘Negro’ among others. Other studies have shown that there is a difference in the content of these nutraceutical compounds between the seed coat and the whole grain (Aquino-Bolaños et al., 2016). These authors disclosed results lower than those found in the present work in seed samples from 26 common bean populations collected in several rural communities in the states of Oaxaca, Guerrero, Puebla, Tlaxcala and Estado de México, presenting a range in whole seed from 0.10 to 0.78 mg CE g-1. Rodriguez et al. (2021) observed that color influences seed flavonoid content in genotypes of Spanish origin, with those of white coat having the lowest amount and those of red color the highest, which coincides with the results obtained in this study with the ‘Sangre de Toro’ variety. Other lines that showed high concentrations of these nutraceutical compounds were pink, brown and black, which does not coincide with the results found in the present work. Anthocyanins constitute one of the most important groups of natural pigments and are responsible for many of the colors of fruits and vegetables, as well as flowers; in beans they are present in higher amounts in black or blue-violet seeds (Guevara-Lara et al., 2006). Anthocyanins in the samples analysed ranged from 0.763 to 2.400 mg C3G g-1. No significant difference was found between ‘Pinto’ and ‘Negro’/104 beans, which had the lowest concentration of these compounds, while ‘Sangre de Toro’, ‘Peruano’ and ‘Flor de Mayo’ showed the highest values with no statistical difference between them. Results were higher than those shown by Guevara-Lara et al. (2006) in wild bean and weed samples from Chiapas, Chihuahua, Durango, Guerrero, Jalisco, Michoacán, Morelos, Nayarit, Oaxaca, Sinaloa and Zacatecas. Likewise, values obtained also exceeded the results presented by Reynoso-Camacho et al. (2007) in ‘Pinto Zapata’, ‘Flor de Mayo’, ‘Anita’ and ‘White Tlaxcala’ beans, but were lower than those obtained in the ‘Flor de Junio Marcela’ variety. They were also higher than the values reported by Herrera-Hernández et al. (2018) in several common bean varieties grown in the state of Zacatecas Mexico. Differences in the content of these bioactive compounds are due to factors such as genotype and place of origin. In contrast to the present study, Rodriguez et al. (2021) found the highest concentration of monomeric anthocyanins in black coat bean samples with an average of 4.40 mg C3G g-1. This Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 17 same result was obtained by Salinas-Moreno et al. (2005) in 15 Mexican black bean varieties. Whereas Aquino- Bolaños et al. (2016) stated that the highest concentration of this bioactive compound was present in the cream-pinkish varieties, with similarity to that obtained in ‘Flor de Mayo’ variety of the current study. Meaning that the color may be independent of the anthocyanin content in the common bean. In summary, of all the varieties analysed, ‘Negro’/104 beans had the lowest concentration of total phenols, flavonoids and anthocyanins, while ‘Flor de Mayo’ and ‘Sangre de Toro’ had the highest values. Table 8Table 8Table 8Table 8. Nutraceutical compounds of common bean varieties produced in Morelos State, Mexico Variety/ Classification number* Total phenols (mg EAG g-1) Phytic acid (g100-1 g-1) Flavonoids (mg CE g-1) Anthocyanins (mg C3G g-1) ‘Sangre de Toro’/98 2.89 ± 0.40ab 0.18 ± 0.009a 1.579 ± 0.10a 2.400 ± 0.11a ‘Peruano’/100 2.24 ± 0.83b 0.182 ± 0.02a 0.275 ± 0.07b 2.147 ± 0.22a ‘Flor de Mayo’/101 4.24 ± 0.57a 0.170 ± 0.01a 1.991 ± 0.09a 2.134 ± 0.18a ‘Negro’/102 1.97 ± 0.57b 0.194 ± 0.08a 0.518 ± 0.12b 1.599 ± 0.07b ‘Pinto’/103 2.19 ± 0.28b 0.175 ± 0.05a 0.638 ± 0.27b 0.763 ± 0.15c ‘Negro’/104 1.76 ± 0.26b 0.149 ± 0.01a 0.416 ± 0.16b 1.133 ± 0.20c Data correspond to mean ± SD. Different letters per column indicate significant statistical difference between varieties. Tukey test (p ≤ 0.05). *Laboratory work classification Correlation analysis Table 9 shows the Pearson correlation coefficients obtained between color variables, nutraceutical compounds and antioxidant ability. Table 9.Table 9.Table 9.Table 9. Pearson correlation coefficients between color variables, nutraceutical compounds and antioxidant capacity of different varieties of common bean grown in Morelos State, Mexico. L a* b* Chroma °Hue AF AC TF Flavono- ids Antho- cyanins L 1 a* -0.13 1 b* 0.92** -0.13 1 Chroma 0.69* 0.42 0.82** 1 °Hue 0.70* 0.29 0.58* 0.60* 1 AF 0.05 0.03 0.08 0.10 0.15 1 AC -0.38 0.44 -0.56* -0.36 -0.01 0.11 1 TF 0.14 0.64* 0.06 0.31 0.34 -0.21 0.50* 1 Flavonoids -0.10 0.83** -0.16 0.23 0.28 0.04 0.61* 0.86** 1 Anthocyanins -0.04 0.54* 0.22 0.57* 0.08 0.10 -0.08 0.429 0.49* 1 *Significant linear correlation (p < 0.05); **highly significant linear correlation (p < 0.0001). °Hue = Hue angle. FA = phytic acid. AC = antioxidant capacity. TF = total phenols. Brightness showed a highly significant positive correlation with b* value (r=0.92), as well as significant with chroma (r=0.69) and hue angle (r=0.70). Likewise, a* value was highly positively correlated with flavonoids (r=0.83) and significantly with total phenols (r=0.64) as well as anthocyanins (r=0.54). As for b* value, it showed a highly significant positive linear correlation with chroma (r=0.82) and significant with hue angle (r=0.58), it also had a significant negative correlation (r=-0.56) with antioxidant capacity. On the other hand, chroma correlated positively and significantly with hue angle (r=0.60) and anthocyanins (r=0.60). While phytic acid showed no correlation with any of the color variables and nutraceutical compounds evaluated; total Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 18 phenols were highly positively correlated with flavonoids (r=0.86) and significantly correlated with antioxidant capacity (r=0.50) whereas flavonoids were significantly and positively correlated with anthocyanins (r=0.49). The results of the present investigation revealed an important contribution of nutraceutical compounds such as total phenols, flavonoids and anthocyanins in the a* value of common bean grown in the producing region of Morelos State, Mexico. Some authors have suggested that the color of the coat of this crop is closely related to the content of several phenolic compounds, prominent among them being flavonoids (Yang et al., 2018) Likewise, in the current work a good correlation was observed between total phenols and flavonoids with antioxidant capacity, this result has been obtained by other authors (Rodriguez et al., 2021; Mastura et al., 2017). However, contrary to what was obtained by other authors (Aquino-Bolaños et al., 2016), in this study no linear correlation (p > 0.05) was observed between anthocyanins and antioxidant capacity. ConclusionsConclusionsConclusionsConclusions The current research showed that genotype has a significant effect on antioxidant capacity, and nutraceutical compound content in bean seed. Obtained colour values coincided with the coloration perceived in the different varieties evaluated. Thus, lower lightness was observed in varieties with darker colours such as ‘Negro’/102, ‘Negro’/104 and ‘Sangre de Toro’ as well as higher lightness in lighter coloured varieties such as ‘Peruano’ bean, ‘Pinto’ and ‘Flor de Mayo’. Furthermore, the highest a* value corresponded to the varieties with a greater tendency to red coloration, such as ‘Sangre de Toro’, and the highest b* value corresponded to those with a greater tendency to yellow coloration, such as ‘Peruano’. On the other hand, the relationship between color and nutraceutical compound content was not very clear in the present study, since the only component that had a significant positive correlation with these was the a* value, which may indicate that the content of these compounds in the seed depends more on the genotype than on the seed coat color. In terms of nutritional and nutraceutical quality, of all the varieties studied, ‘Flor de Mayo’ beans had the highest crude fiber content, total phenols, flavonoids, anthocyanins and antioxidant ability, and, after ‘Peruano’ beans, the highest protein content. The correlation found between flavonoids and total phenols with antioxidant capacity shows these nutraceutical compounds as important indicators of antioxidant capacity of the seeds evaluated. Finally, the study showed that the bean varieties produced in Morelos State, Mexico, may be of interest from a functional and nutritional point of view. Likewise, these varieties are favourable lines to be biofortified or used in genetic improvement programs in the future, for benefiting the population whose basic source of protein is this legume or those with scarce resources. Authors’ ContributionsAuthors’ ContributionsAuthors’ ContributionsAuthors’ Contributions Both authors read and approved the final manuscript. Ethical approvalEthical approvalEthical approvalEthical approval (for researches involving animals or humans) Not applicable. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 19 AcknowledgementsAcknowledgementsAcknowledgementsAcknowledgements We thank to Consejo Nacional de Ciencia y Tecnología (CONACyT)-México (National Council of Science and Technology, Mexico) for the support granted in the "Convocatoria Proyectos de Desarrollo Científico para Atender Problemas Nacionales 2015" (Call for Scientific Development Projects to Address National Problems 2015), to Project 1529 "Biofortificación de cultivos agrícolas básicos, clave para combatir la desnutrición" (Biofortification of basic agricultural crops, key to fight malnutrition). Conflict ofConflict ofConflict ofConflict of InterestsInterestsInterestsInterests The authors declare that there are no conflicts of interest related to this article. ReferencesReferencesReferencesReferences Aguilera Y, Estrella I, Benitez V, Esteban RM, Martín-Cabrejas MA (2011). Bioactive phenolic compounds and functional properties of dehydrated bean flours. Food Research International 44:774-780. https://doi.org/10.1016/j.foodres.2011.01.004 Aguirre SEA, Gómez-Aldapa CA (2010). Evaluación de las características fisicoquímicas en la especie de frijol Phaseolus vulgaris de las variedades Pinto Saltillo, Bayo Victoria y Negro San Luis [Evaluation of the physicochemical characteristics in the bean species Phaseolus vulgaris of the varieties Pinto Saltillo, Bayo Victoria and Negro San Luis]. In: Proceedings of the XII Congreso Nacional de Ciencia Y Tecnología de Alimentos, Guanajuato, México pp 27-28. Aguirre-Santos EA, Rodríguez-Miranda J, Rosales-Serna R, Castro-Rosas J, Ochoa-Martínez LA, Valle-Cervantes S, Gómez-Aldapa CA (2011). Determinación de tiempos de cocción de frijol común var. Pinto Saltillo utilizando dos métodos (Determination of cooking times of common bean variety Pinto Saltillo using two methods). In: Proceedings of the XXXII Encuentro Nacional y 1er Congreso Internacional AMIDIQ, Riviera Maya, Quintana Roo pp 2812-2818. Akond AGM, Khandaker L, Berthold J, Gates L, Peters K, Delong H, Hossain K (2011a). Anthocyanin, total polyphenols and antioxidant activity of common bean. American Journal of Food Technology 6(5):85-394. https://doi.org/10.1016/j.foodchem.2009.01.037 Akond AGM, Crawford H, Berthold J, Talukder ZI, Hossain K (2011b). Minerals (Zn, Fe, Ca and Mg) and antinutrient (phytic acid) constituents in common bean. American Journal of Food Technology 6(3):235. https://doi.org/10.3923/ajft.2011.235.243 Aquino-Bolaños EN, García-Díaz YD, Chavez-Servia JL, Carrillo-Rodríguez JC, Vera-Guzmán AM, Heredia-García E (2016). Anthocyanin, polyphenol, and flavonoid contents and antioxidant activity in Mexican common bean (Phaseolus vulgaris L.) landraces. Emirates Journal of Food and Agriculture 28:581-588. https://doi.org/10.9755/ejfa.2016-02-147 Araméndiz-Tatis H, Cardona-Ayala CE, Combatt-Caballero EM (2016). Contenido nutricional de líneas de fríjol caupí (Vigna unguiculata L. Walp.) seleccionadas de una población criolla [Nutritional content of cowpea bean lines (Vigna unguiculata L. Walp.) selected from a native population]. Información Tecnológica 27(2):53-60. http://dx.doi.org/10.4067/S0718-07642016000200007 Armendáriz-Fernández KV, Herrera-Hernández IM, Muñoz-Márquez E, Sánchez E (2019). Characterization of bioactive compounds, mineral content, and antioxidant activity in bean varieties grown with traditional methods in Oaxaca, Mexico. Antioxidants 8(1):26. https://doi.org/10.3390/antiox8010026 Association of Official Analytical Chemist (AOAC) (2000). Official Methods of Analysis (17th). Ed. AOAC International. Guithersbur, MD, EE. UU. Beninger CW, Hosfield G, Bassett M (1999). Flavonoid composition of three genotypes of dry bean (Phaseolus vulgaris) differing in seedcoat color. Journal of the American Society for Horticultural Science 124(5):514-518. Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 20 Cardador-Martínez A, Loarca-Piña G, Oomah BD (2002). Antioxidant activity in common beans (Phaseolus vulgaris L.). Journal of Agricultural and Food Chemistry 50:6975-6980. https://doi.org/10.1021/jf020296n Chávez-Mendoza C, Hernández-Figueroa KI, Sánchez E (2019). Antioxidant capacity and phytonutrient content in the seed coat and cotyledon of common beans (Phaseolus vulgaris L.) from various regions in Mexico. Antioxidants 8(1):5. https://doi.org/10.3390/antiox8010005 De-Paula CD, Jarma-Arroyo S, Aramendiz-Tatis H (2018). Caracterización nutricional y determinación de ácido fítico como factor antinutricional del frijol caupí. [Nutritional characterization and determination of phytic acid as an antinutritional factor in cowpea beans.] Agronomía Mesoamericana 29(1):30-41. http://dx.doi.org/10.15517/ma.v29i1.27941 Díaz AM, Caldas GV, Blair MW (2010). Concentrations of condensed tannins and anthocyanins in common bean seed coats. Food Research International 43(2):595-601. https://doi.org/10.1016/j.foodres.2009.07.014 Díaz-Batalla L, Widholm JM, Fahey GC, Castaño-Tostado E, Paredes-López O (2006) Chemical components with health implications in wild and cultivated Mexican common bean seeds (Phaseolus vulgaris L.). Journal of Agricultural and Food Chemistry 54:2045-2052. https://doi.org/10.1021/jf051706l Espinosa-Alonso LG, Lygin A, Widholm JM, Valverde ME, Paredes-Lopez O (2006). Polyphenols in wild and weedy Mexican common beans (Phaseolus vulgaris L.). Journal of Agricultural and Food Chemistry 12:4436-4444. https://doi.org/10.1021/jf060185e Espinoza-García N, Martínez-Martínez R, Chávez-Servia JL, Vera-Guzmán AM, Carrillo-Rodríguez JC, Heredia-García E, Velasco-Velasco VA (2016). Contenido de minerales en semilla de poblaciones nativas de frijol común (Phaseolus vulgaris L.) [Mineral content in seeds of native populations of common bean (Phaseolus vulgaris L.)]. Revista Fitotecnica Mexic 3:215-223. Fernández VAF, Sánchez ChE (2017). Estudio de las propiedades fisicoquímicas y calidad nutricional en distintas variedades de frijol consumidas en México [Study of physicochemical properties and nutritional quality in different varieties of beans consumed in Mexico]. Nova Scientia 9(18):133-148. Gálvez RL, Genovese MI, Lajolo FM (2007). Polyphenols and antioxidant capacity of seed coat and cotyledon from Brazilian and Peruvian bean cultivars (Phaseolus vulgaris L.). Journal of Agricultural and Food Chemistry 1:90- 98. https://doi.org/10.1021/jf062785j González de Mejía EG, Castaño-Tostado E, Loarca-Piña G (1999). Antimutagenic effects of natural phenolic compounds in beans. Mutation Research - Genetic Toxicology and Environmental Mutagenesis 441:1-9. https://doi.org/10.1016/S1383-5718(99)00040-6 Guevara-Lara F, Espinosa-Alonso LG, Valverde ME, Lygin A, Widholm J, Paredes-López O (2006). Phenolics, flavonoids and other nutraceuticals in Mexican wild common beans (Phaseolus vulgaris). In: Michael H. Tunick, Elvira González de Mejia (Eds). Hispanic Foods Chemistry and Flavor. ACS Symposium Series 946:77-88. https://doi.org/10.1021/bk-2007-0946.ch007 Gupta RK, Gangoliya SS, Singh NK (2015). Reduction of phytic acid and enhancement of bioavailable micronutrients in food grains. Journal of Food Science and Technology 52(2):676-684. https://doi.org/10.1007/s13197-013-0978-y Herrera-Hernández IM, Armendáriz-Fernández KV, Muñoz-Márquez E, Sida-Arreola JP, Sánchez E (2018). Characterization of bioactive compounds, mineral content and antioxidant capacity in bean varieties grown in semi-arid conditions in Zacatecas, Mexico. Foods 7(12):199. https://doi.org/10.3390/foods7120199 Huber K, Brigide P, Bretas EB, Canniatti-Brazaca SG (2016) Phenolic acid, flavonoids and antioxidant activity of common brown beans (Phaseolus vulgaris L.) before and after cooking. Journal of Nutrition & Food Sciences 6:551. https://doi.org/10.4172/2155-9600.1000551 Hsu CL, Chen W, Weng YM, Tseng CY (2003). Chemical composition, physical properties, and antioxidant activities of yam flours as affected by different drying methods. Food Chemistry 83:85-92. https://doi.org/10.1016/S0308- 8146(03)00053-0 Iniestra-González JJ, Ibarra-Pérez FJ, Gallegos-Infante JA, Rocha-Guzmán NE, González-Laredo RF (2005). Factores antinutricios y actividad antioxidante en variedades mejoradas de frijol común (Phaseolus vulgaris) [Antinutritional factors and antioxidant activity in improved varieties of common bean (Phaseolus vulgaris)]. Agrociencia 39: 603-610. Martinez VE (2016). El calcio, esencial para la salud. [Calcium, essential for health]. Nutricin Hospitalaria 33:26-31. http://dx.doi.org/10.20960/nh.341 Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 21 Mastura HY, Hasnah H, Dang TN (2017). Total phenolic content and antioxidant capacity of beans: Organic vs. inorganic. International Food Research Journal 24:510-517. McGuire RG (1992). Reporting of objective color measurements. HortScience 12:1254-1255. https://doi.org/10.21273/HORTSCI.27.12.1254 McKie VA, McCleary BV (2016). A novel and rapid colorimetric method for measuring total phosphorus and phytic acid in foods and animal feeds. Journal of AOAC International 99(3):738-743. https://doi.org/10.5740/jaoacint.16- 0029 Mederos Y, Reynaldo IM (2007). Determinación de indicadores de calidad en 11 genotipos de la especie Phaseolus vulgaris, L. [Determination of quality indicators in 11 genotypes of the species Phaseolus vulgaris L.] Cultivos Tropicales, 28(4):51-56. Retrieved 2022 October 12 from: https://www.redalyc.org/articulo.oa?id=193217894007 Mederos Y (2006). Indicadores de la calidad en el grano de frijol (Phaseolus vulgaris L.) [Quality indicators in bean grain (Phaseolus vulgaris L.)]. Cultivos Tropicales, 27(3):55-62. Retrieved 2022 September 18 from: https://www.redalyc.org/pdf/1932/193215825009.pdf Messina V (2014). Nutritional and health benefits of dried beans. The American Journal of Clinical Nutrition 100:437S- 442S. https://doi.org/10.3945/ajcn.113.071472 Mudryj AN, Yu N, Aukema HM (2014). Nutritional and health benefits of pulses. Applied Physiology, Nutrition, and Metabolism 39(11):1197-1204. https://doi.org/10.1139/apnm-2013-0557 NOM-051-SCFI/SSA1-2010 (2010). Especificaciones Generales de Etiquetado Para Alimentos Y Bebidas No Alcohólicas Preenvasados-Información Comercial Y Sanitaria [General Labeling Specifications for Prepackaged Food And Non-Alcoholic Beverages-Commercial And Sanitary Information]. Retrieved 2022 September 21 from: http://dof.gob.mx/nota_detalle.php?codigo=5137518&fecha=05/04/2010 NMX-F-066-S-1978 (1978). Determinación de cenizas en alimentos [Determination of ash in food]. Normas Mexicanas. Dirección General de Normas. Retrieved 2022 October 23 from: http://www.colpos.mx/bancodenormas/nmexicanas/NMX-F-066-S-1978.PDF NOM-F-90-S-1978 (1978). Determinación de fibra cruda en Alimentos [Determination of crude fiber in Food]. Norma Oficial Mexicana. Retrieved 2022 October 23 from: http://www.dof.gob.mx/nota_detalle.php?codigo=479984&fecha=27/03/1979 Paredes M, Becerra V, Tay J (2009). Inorganic nutritional composition of common bean (Phaseolus vulgaris L.) genotypes race Chile. Chilean Journal of Agricultural Research 69:486-495. http://dx.doi.org/10.4067/S0718- 58392009000400002. Pérez LH (2006). Nutracéuticos: componente emergente para el beneficio de la salud [Nutraceuticals: emerging component for health benefit]. ICIDCA. Sobre los Derivados de la Caña de Azúcar 40(3):20-28 Peña-Betancourt SD, Conde-Martínez V (2012). Aflatoxin and protein content in 13 bean (Phaseolus vulgaris L.) varieties. Revista mexicana de Ciencias Agrícolas 3(1):201-206. Pliego-Marín L, López-Baltazar J, Aragón-Robles E (2013). Características físicas, nutricionales y capacidad germinativa de frijol criollo bajo estrés hídrico [Physical, nutritional and germination characteristics of native beans under water stress]. Revista Mexicana de Ciencias Agrícolas 4(SPE6):1197-1209. Reussi-Calvo NI, Echeverría HE, Sainz-Rozas H (2008). Comparación de métodos de determinación de nitrógeno y azufre en planta: Implicancia en el diagnóstico de azufre en trigo [Comparison of methods for determining nitrogen and sulfur in plants: Implication in the diagnosis of sulfur in wheat]. Ciencia Del Suelo 26(2):161-167. Reynoso-Camacho R, Ríos-Ugalde MC, Torres-Pacheco I, Acosta-Gallegos JA, Palomino-Salinas AC, Ramos-Gómez M, … Guzmán-Maldonado SH (2007). Common bean (Phaseolus vulgaris L.) consumptions and its effects on colon cancer in Sprague Dawley rats. Agricultura Técnica en México 33(1):43-52. Rodríguez MR. Campa-Negrillo A, Suárez-Valles B, Ferreira-Fernández JJ (2021). Phenolic content and antioxidant activity in seeds of common bean (Phaseolus vulgaris L.). Foods 10:864. https://doi.org/10.3390/foods10040864 Rodríguez-Blanco L, Lucas-Florentino B, Miranda-Cruz E, Guerrero-Olazarán M (2018). Contenido de ácido fítico y fósforo inorgánico en Vigna unguiculata y Phaseolus vulgaris germinadas a diferentes temperaturas [Content of phytic acid and inorganic phosphorus in Vigna unguiculata and Phaseolus vulgaris germinated at different temperatures]. Información Tecnológica 29(3):39-46. http://dx.doi.org/10.4067/S0718-07642018000300039 Chávez-Mendoza C and Sánchez E (2023). Not Sci Biol 15(1):11353 22 Salinas-Moreno Y, Rojas-Herrera L, Sosa-Montes E, Pérez-Herrera P (2005). Composición de antocianinas en variedades de firijol negro (Phaseolus vulgaris L.) cultivadas en México [anthocyanin composition in black bean (Phaseolus vulgaris L.) varieties grown in México]. Agrociencia 39:385-394. Silva-Cristobal L, Osorio-Díaz P, Tovar J, Bello-Pérez LA (2010). Chemical composition, carbohydrate digestibility, and antioxidant capacity of cooked black bean, chickpea, and lentil Mexican varieties. CyTa-Journal of Food 1:7-14. https://doi.org/10.1080/19476330903119218 Singleton VL, Rossi JA (1965). Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagents. American Society for Enology and Viticulture 16:144-158. Retrieved 2022 September 23 from: https://www.ajevonline.org/content/16/3/144 Suárez-Martínez SE, Ferriz-Martínez RA, Campos-Vega R, Elton-Puente JE, de la Torre Carbot K, García-Gasca T (2016) Bean seeds: leading nutraceutical source for human health, CyTA-Journal of Food 14:131-137 https://doi.org/10.1080/19476337.2015.1063548 Takeoka GR, Dao LT, Full GH, Wong RY, Harden LA, Edwards RH, Berrios S (1997). Characterization of black bean (Phaseolus vulgaris L.) anthocyanins. Journal of Agricultural and Food Chemistry 45:3395-3400. https://doi.org/10.1021/jf970264d Taruscio TG, Barney DL, Exon J (2004). Content and profile of flavonoid and phenolic acid compounds in conjunction with the antioxidant capacity or a variety of Northwest Vaccinium narreies. Journal of Agricultural and Food Chemistry 52:3169-3176. https://doi.org/10.1021/jf0307595 Ulloa JA, Ulloa RP, Ramírez-Ramírez JC, Ulloa-Rangel BE (2011). El frijol (Phaseolus vulgaris): su importancia nutricional y como fuente de fitoquímicos [The bean (Phaseolus vulgaris): its nutritional importance and as a source of phytochemicals]. Revista Fuente 3(8):5-9. Wrolstad RE, Durst RW, Lee J (2005). Tracking color and pigment changes in anthocyanin products. Trends in Food Science & Technology 16(9):423-428. https://doi.org/10.1016/j.tifs.2005.03.019 Yang Q Q, Gan RY, Ge YY, Zhang D, Corke H (2018). Polyphenols in common beans (Phaseolus vulgaris L.): Chemistry, analysis, and factors affecting composition. Comprehensive Reviews in Food Science and Food Safety 17(6):1518- 1539. https://doi.org/10.1111/1541-4337.12391 Zhishen J, Mengcheng T, Jianming W (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry 64:555-559. https://doi.org/10.1016/S0308-8146(98)00102-2 The journal offers free, immediate, and unrestricted access to peer-reviewed research and scholarly work. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author. 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