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TECNOCIENCIA CHIHUAHUA, Vol. XIX (1): e1691 (enero-abril. 2025)
https://vocero.uach.mx/index.php/tecnociencia
ISSN-e: 2683-3360
Artículo Científico
Variation of the seed and foliar phenolic contents
of five wild forms of common bean (Phaseolus
vulgaris L.)
Variación del contenido fenólico foliar y de semillas de cinco
formas silvestres de frijol común (Phaseolus vulgaris L.)
*Correspondencia: lwallanderc@ipn.mx (Liliana Wallander-Compeán)
DOI: https://doi.org/10.54167/tch.v19i1.1691
Recibido: 08 de octubre de 2024; Aceptado: 20 de enero de 2025
Publicado por la Universidad Autónoma de Chihuahua, a través de la Dirección de Investigación y Posgrado.
Editora de Sección: Dra. Yolanda Salinas-Moreno
Abstract
The objective of the present study was to evaluate the seed and foliar phenolic composition from
populations of wild Phaseolus vulgaris of Durango, Mexico. Seeds and leaves extracts were analyzed
by HPLC-DAD, and UV-visible spectrophotometry. Analysis of variance were used to determine the
capacity of the phenolic contents to discriminate between samples, the data were submitted to
principal component analysis (PCA) and cluster analysis. The population with the highest content
of phenolic compounds in both seed and leaves was the wild population of Nuevo Ideal and those
that accumulated the lowest content of these compounds were Nombre de Dios and Canatlán. A
total of 37 phenolic compounds in both seed and leaves from wild common bean were identified by
HPLC–DAD. Wild species are important for the conservation of biodiversity, and for the genetic
improvement of new varieties. Likewise, they could be used as forage, food or medicine, due to the
high content of phytochemicals in seeds and leaf tissue, therefore, the information generated is
relevant to knowledge about the characterization and diversity of wild Phaseolus species.
Keywords: phytochemicals, wild forms, spectroscopy, Phaseolus vulgaris, HPLC-DAD, phenolic
composition.
Shaila Nayeli Pérez-Salinas1, Rene Torres-Ricario1, Nancy Nohemí Rodarte-Rodríguez 1,
Heberto Iván Salas-Ayala1, Liliana Wallander-Compeán1*
1 Instituto Politécnico Nacional CIIDIR Unidad Durango, Av. Sigma 119, Fraccionamiento 20 de
noviembre II, Durango, Durango. C.P.34220.
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Resumen
El objetivo del presente estudio fue evaluar la composición fenólica de hojas y de semillas de
poblaciones silvestres de Phaseolus vulgaris de Durango, México. Los extractos de semillas y hojas se
prepararon y analizaron mediante HPLC-DAD y espectrofotometría UV-visible. Se utilizó análisis
de varianza para determinar la capacidad de los contenidos fenólicos para discriminar entre
muestras. Los datos se sometieron a análisis de componentes principales (PCA) y análisis de
agrupamiento. La población con mayor contenido de compuestos fenólicos tanto en semilla como en
hoja es la población silvestre de Nuevo Ideal y las que acumularon menor contenido de estos
compuestos fueron Nombre de Dios y Canatlán. Mediante HPLC-DAD se identificaron un total de
37 compuestos fenólicos tanto en semilla como en hojas de frijol común silvestre. Las especies
silvestres son importantes para la conservación de la biodiversidad, y para el mejoramiento genético.
Asimismo, podrían usarse como forraje, alimento o medicina, debido al alto contenido de
fitoquímicos en tejido foliar y semillas. Por lo tanto, la información generada es relevante para el
conocimiento sobre la caracterización y diversidad de especies silvestres de Phaseolus.
Palabras clave: fitoquímicos, formas silvestres, espectroscopia, Phaseolus vulgaris, HPLC-DAD,
composición fenólica.
1. Introduction
Legumes are important sources of nutrients and bioactive compounds. Depending on the
genotype, bean seeds also contain a wide variety of polyphenolic compounds, which have
prospective health benefits. These may include flavonoids such as anthocyanins, flavonoids,
proanthocyanidins, and tannins, as well as a wide range of phenolic acids (Ganesan and Xu, 2017;
Yang et al., 2018). Phenolic compounds contribute to the taste, smell and color of foods, and their
bioactive role as antioxidants is associated with the prevention of cardiovascular and
neurodegenerative diseases, diabetes, and cancer (Bhuyan and Basu, 2017).
The analysis of the foliar phenolic composition of common bean has received less attention than that
of seeds, probably because humans do not consume the leaves. However, as leaves of common bean
are used as forage for livestock, it is important to determine their phenolic composition because
phenolics have nutraceutical properties (Mueller et al., 2019). This may improve not just animal
health, but the meat quality could be enhanced, and even organic waste improves soil structure.
All the previous studies worked just with domesticated beans and were focused on plant phenolic
contents. These reports registered a variation according to changes in the environmental growth
conditions (Del Valle et al., 2015), plant age (Bystricka et al., 2014), and genotypes (Yusnawan et al.,
2018). The state of Durango has physiographic and climatic diversity, due to this diversity wild
species of common bean develop with variable phenotypes. In the last ten years the state has
occupied the second place with the highest production of this legume at the national level. Wild
species represent important information for the improvement and development of new varieties, so
it is necessary to protect and conserve this plant genetic resource.
Hence, it is important to determine the seed and foliar phenolic contents among varieties and wild
forms of common bean growing in different environmental conditions. There are few reports on the
variation in foliar phenolic content of domesticated common bean (Reyes et al., 2014). The leaf tissue
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and seeds come from different municipalities with different altitudes, El Mezquital located at a low
elevation, Súchil and Nombre de Dios at mid elevation, and Canatlán and Nuevo Ideal at high
elevations. In Mexico there are few previous reports on the phenolic contents of wild forms of P.
vulgaris. The objective of this study was to determine and compare the phenolic content of seeds and
leaf tissue of five wild forms of common bean from Durango, Mexico.
2. Materials and methods
2.1 Materials
2.1.1 Plant Materials
Seeds (Fig. 1) and leaves of five wild forms of common bean were collected from their natural
populations in Durango, Mexico (Table 1) during september - december 2019. The leaves and seeds
were dried in a botanical drier at 40 °C to constant weight, ground in a domestic blender, and stored
in paper bags in dark at room temperature.
Table 1. Collection data for five wild common beans (Phaseolus vulgaris) from Durango, Mexico.
Tabla 1. Datos de recolección de cinco frijoles silvestres (Phaseolus vulgaris) de Durango, México.
Localities
Latitude N
Longitude W
Altitude
(m)
Weather
Tmax/Tmin*
(°C)
El Mezquital
23° 26´ 48.1´´
104° 21´ 49.5´´
1400
Temperate
subhumid
42.4/-0.2
Súchil
23° 39´24.7´´
104° 02´ 20.9´´
1963
Temperate
35.7/-6.4
Nombre de Dios
24° 04’ 71’’
104°14´ 23´´
1877
Temperate
36.7/-3.5
Canatlán
24° 51´03.4´´
104°51´44.8´´
2039
Semicold
35.1/-7.3
Nuevo Ideal
24° 45´11.9´´
105° 00´ 05.6´´
2037
Temperate
36.2/-5.0
*Data obtained from CONAGUA (2019).
Figure 1. Localities provenance and pictures of the seeds of five populations of wild common beans
(Phaseolus vulgaris) from Durango, Mexico.
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Figura 1. Localidades de procedencia y fotografías del grano de cinco poblaciones de frijol común silvestre
(Phaseolus vulgaris) de Durango, México.
2.2 Methods
2.2.1 Preparation of extracts
Phenolic extracts were prepared according to Kim and Lee (2002). One gram of dry ground
samples was macerated in 10 mL of 80 % methanol (v/v) for 24 hours, in the dark at room
temperature. The extracts were centrifuged at 1,013 g, in a BOECO centrifuge U-32R, 230-240V, 50/60
Hz, for ten minutes at room temperature and the supernatants were separated. The pellets were
reextracted and macerated in 10 mL of 20 % methanol (v/v) for one hour and centrifuged under the
same conditions. Both solvents from the same sample were combined and formed the total extract,
which was concentrated to dryness (by evaporation) and resuspended in 80 % methanol. For the
HPLC-DAD analysis of both foliar and seed samples, total extracts were concentrated to dryness (by
evaporation), and dissolved in 3 mL of methanol; these were the phenolic enriched extracts.
2.2.2 Spectrometric analysis of Phenolic compounds
The equipment used was the Multiskan Go spectrophotometer; high-quality
monochromatorbased UV/VIS spectrophotometer. It is used in spectral scanning, endpoint and
kinetic analysis. It measures absorbance in the 200–1000 nm wavelength range and it appropriate to
96- or 384-well plates.
Total Phenolic content (TPC)
The total phenolic concentrations were determined according to Falleh et al. (2011), from a
standard curve constructed with six concentrations of gallic acid (Abs 760nm = 4.4875 [gallic acid] +
0.0368, correlation coefficient r = 0.9946). Total phenolics concentrations were expressed as
milligrams of gallic acid equivalents per gram of dry weight (mg GAE / 100 g DW).
Flavonoid contents (FC)
The flavonoid contents were determined according to Lauranson and LebGreton (1993),
from a standard curve constructed with six concentrations of quercetin (Abs 420 nm = 6.7545
[quercetin] + 0.081, correlation coefficient r = 0.997). Flavonoid contents were expressed as milligrams
of quercetin equivalents per 100 grams of dry weight (mg QE / 100 g DW).
Total condensed tannins (TCT)
The contents of condensed tannins were determined according to Julkunen (1985), from a
standard curve constructed with seven concentrations of epicatechin (Abs 510 nm = 0.0113
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[epicatechin] – 0.0055 r = 0.9982). The contents were expressed as milligrams of epicatechin
equivalents per gram of dry weight (mg EC / 100 g DW).
Anthocyanins (AC)
As reference, Monomeric anthocyanins were determined with the differential pH method
described by Giusti and Wrolstad (2001), Abs 510 nm y Abs 700 nm using the molecular weight
(449.2) and the molar absorption (26.900) of cyanidin-3-O-glucoside as reference. The values were
expressed as milligrams of cyanidin-3-O-glucoside per dry extract (mg C3OG / 100 g DE).
2.3 HPLC-DAD analysis
Aliquots (100 uL) of foliar and seeds phenolic enriched extracts (50 mg/mL) were analyzed
in a Perkin Elmer® Flexar™ PDA Plus™ detector, according to the gradient method reported by da
Graça and Markham (2007). Chromatographic conditions: Mobile phase: A: acidified water
(phosphoric acid) y B: acetonitrile, 20 uL of sample was injected, with a flow of 0.8 uL/min. Column:
Brownlee™ SPP C18, 100 x 2.1 mm, 2.7 µm at 45 °C. Chromatograms were registered at 265 and 340
nm. Spectral data of all picks were recorded between 200 and 400 nm using diode array detection
(DAD).
2.4 Experimental design and data analysis
The data of phenolic contents were subjected to an analysis of variance (p ≤ 0.05) and means
were compared with the Tukey test. To determine the capacity of the phenolic contents to
discriminate between samples, the data were submitted to principal component analysis (PCA) and
cluster analysis, XLSTAT v 2021.2.2 (Addinsoft 2024).
3. Results and discussion
3.1 Spectrometric analysis of phenolic compounds
3.1.1 Phenolic compounds in seeds
The seeds accumulated different pigments in the coat (Fig. 1). In common bean seeds, pigments
are predominantly phenolics (Capistrán et al., 2019; Rodriguez et al., 2021). The coloration of the
common bean seed is related to the P (pigment) gene and, in the case of the white coloration in the
coat seed, it is due to a recessive phenotype of all the races that have been domesticated of this species
along its history. Moreover, the same pigmentation of the seed has been used as a quantitative trait
genetic marker (McClean et al., 2018).
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Total Phenolic content
Significant variations were found in the content of secondary metabolites in seeds (Table 2). The
locality of Nuevo Ideal found higher TPC (269.9 mg GAE/ 100 g DW), and El Mezquital TPC 173.67
mg GAE / 100 g DW, in the locality Nombre de Dios the lowest amount was found of (133.39 mg
GAE / 100 g DW). The main beneficial health effect of phenolics is their antioxidant activity (Khang
et al., 2016; Singh et al., 2017; Carbas et al., 2020), have reported values of total phenolics in common
beans of 117 mg EAG/100 g to 440 mg EAG/100 g.
Flavonoid contents
The bean populations that presented the highest content of flavonoids was from the locality of
Nuevo Ideal (77.66 mg QE / 100 g DW) and the lowest amount was found in the town of El Mezquital
(50.33 mg QE / 100 g DW). Previous studies have reported that black bean varieties are characterized
by having the highest flavonoid values compared to light varieties (Armendáriz et al., 2019). The
nutraceutical properties attributed to the bean have been mainly related to this type of bioactive
compounds. The seeds from the locality of Nuevo Ideal is a variegated seed with dark colors,
between black and grey. It has been reported that anthocyanins are responsible for the red, black
and red pigmentation, blue in bean grains, these pigments are generally located in the seed coat of
the grain (Dzomba et al., 2013; Harlen and Jati, 2018). According to Heredia (2017), the consumption
of common beans has adequate concentrations of flavonoids, in addition, when compared with other
legumes (Chickpea 124.59 µg QE / g DW, lentils 17.13 µg QE / g DW) it has a greater amount of this
element. Likewise, when grain consumption is complete, there is a considerable biological effect of
flavonoids in legumes, showing anticancer and anti-inflammatory properties (Maleki et al., 2019).
Within the group of flavonoids, anthocyanins are the most important type, which are found in
pigmented beans and are responsible for the antioxidant capacity.
Total condensed tannins
The concentration of tannins in the wild common bean populations evaluated showed values in
a range of 51.35 mg EC / 100 g DW to 75.17 mg ECE / 100 g DW. Tannins are compounds that can
have a beneficial or adverse nutritional effect and are the predominant phenolic compounds in
legumes. These can interact with macronutrients, especially with proteins (Zhang et al., 2014).
Anthocyanins
The wild forms of common bean studied showed anthocyanin concentrations in a range from
0.38 mg C3OG / 100 g DW to 3.12 mg C3OG / 100 g DE. The highest content of anthocyanins was
found in the seeds from El Mezquital (3.120 mg C3OG / 100 g) and Nuevo Ideal (1.73 mg C3OG / 100
g), seeds with dark colors in the case of those from El Mezquital and those from New Ideal with
marbled colour. Dark coloured beans are considered a good source of anthocyanins. (Mojica et al.,
2017; Harlen 2018).
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Table 2. Contents of total phenolics, flavonoids, condensed tannins, and anthocyanins of seeds of five wild
forms of common bean (Phaseolus vulgaris) from Durango, Mexico.
Tabla 2. Contenidos de fenoles totales, flavonoides, taninos condensados y antocianinas de semillas de cinco
formas silvestres de frijol común (Phaseolus vulgaris) de Durango, México.
Localities
Total phenolics1
Flavonoids2
Condensed tannins3
Anthocyanins4
El Mezquital
173.67 ± 10.39 b
50.33 ± 6.81 b
74.30 ± 1.27 a
3.12 ± 0.11 a
Súchil
160.74 ± 7.50 c
69.81 ± 5.75 ab
71.58 ± 3.33 ab
1.14 ± 0.41 c
Nombre de Dios
133.39 ± 5.64 c
76.56 ± 6.32 a
68.69 ± 1.40 b
0.85 ± 0.11 c
Canatlán
154.20 ± 1.63 c
55.80 ± 9.27 ab
51.35 ± 1.93 c
0.38 ± 0.10 d
Nuevo Ideal
269.90 ± 2.95 a
77.66 ± 8.58 a
75.17 ± 0.66 a
1.73 ± 0.18 b
Pr > F(Model)
0.0.14
0.048
<0.0001
<0.0001
Significant
Yes
Yes
Yes
Yes
1: Expressed in mg GAE / 100 g DW, 2: mg QE / 100 g DW, 3: mg EC / 100 g DW, 4: mg C3OG / 100 g DW.
Different uppercase letters indicate significant differences between five populations (p ≤ 0.05) (Fisher’s test),
media ± SD, n = 3.
In this study, the seeds with the highest pigmentation (dark colors) were those from Nuevo Ideal
and El Mezquital, likely contain a diverse range of beneficial phenolic compounds, contributing to
their higher antioxidant activity, followed by those from Súchil, and the lighter-colored seeds from
Canatlán and Nombre de Dios, may limit their health benefits in comparison to more pigmented
alternatives. According to the literature, the seeds with the highest pigmentation, in this case those
from Nuevo Ideal and El Mezquital, presented the highest levels and the one with the lowest
phenolic content was the seed from Nombre de Dios, the lightest-colored seed included in this study.
Variation in phenolic composition and antioxidant activities between common bean cultivars and
their processed products has also been widely studied, for example, Akon et al., 2011; Suárez et al.,
2015; Aquino et al., 2016. The phenolic profile can vary not just between cultivated varieties, but also
between wild species, which can manifest similarities in composition. However, differences could
occur due to factors like pigmentation (color) and their environmental adaptation, which influences
their biochemical pathways. For instance, colored bean varieties often have higher levels of specific
phenolic compounds compared to lighter colored ones due to the presence of additional pigments
like anthocyanins. (Kleintop et al., 2016; Murube et al., 2021; Campa et al., 2023). The significant
differences among varieties highlight the importance of cultivar selection in breeding programs
aimed at enhancing the nutritional quality of common beans. (Assefa et al., 2019).
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3.2 Phenolic compounds in leaves
Total Phenolic content
Regarding the content of phenolic compounds in leaves, significant variations were found (Table
3). The locality of Canatlán found higher TPC (828.98 mg GAE / 100 g DW), and in the locality of
Nombre de Dios the lowest content was found (145.27 mg GAE / 100 g DW).
Flavonoid contents
The highest concentration in flavonoids was found in leaves population from El Mezquital (212.68
mg QE / 100 g DW) and the leaves populations of Nombre de Dios (48.45 mg QE / 100 g DW) and
Súchil (49.57 mg EC / 100 g DW) were the ones that showed the lowest concentration. Some
environmental factors such as ultraviolet radiation, low temperatures, droughts, water stress, among
others, have been shown to induce the accumulation of anthocyanins in plants (Harlen et al., 2018;
Maleki et al., 2019).
Total condensed tannins
The concentration of tannins in the leaves of the wild common bean evaluated showed values in
a range of 5.20 mg EC / 100g DW to 10.91 mg ECE /100 g DW. The locality of Canatlán was the one
with the highest concentration of condensed tannins (10.911 mg ECE / 100 g DW) and the town of
Nuevo Ideal with the lowest amount (5.20 mg ECE / 100 g DW).
Anthocyanins
The concentration of anthocyanins in the leaves of the wild common bean evaluated showed
values in a range of 3.89 mg C3OG / 100 g DE to11.54 mg C3OG / 100 g DE. The locality of Nuevo
Ideal along with that of Súchil presented higher levels than the rest of the analyzed leaves (11.54 and
10.09 mg C3OG / 100 g DE) respectively, the lowest value was found in the locality Nombre de Dios
3.89 mg C3OG / 100 g DE.
In this study the leaves were taken in the vegetative phase. At this stage, the leaves are usually more
tender and softer. Young leaves, in particular, are rich in nutrients and bioactive compounds, such
as phenolic compounds, which play important roles in defense against pests and diseases, as well as
in the regulation of physiological processes. (Chávez & Sánchez 2017; Gaafar et al., 2020; Šamec et al.,
2021). The leaves in which the greatest amount of phenolic compounds were obtained were those
from Nuevo Ideal, El Mezquital and Canatlán and in which the least amount of these compounds
was found is in the sample from Nombre de Dios, this coincides with what was found in the seeds
analyzed in said study. The studies reported by Torche et al. (2018) have shown that young common
bean leaves can be a valuable source of these compounds, which makes their study very significant
in agrobiology and nutrition. The content of phenolic compounds in wild common bean leaves has
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been little explored when compared to the studies that exist on the content of these same compounds
in its seeds.
Table 3. Contents of total phenolics, flavonoids, condensed tannins, and anthocyanins of leaves of five wild
forms of common bean (Phaseolus vulgaris) from Durango, Mexico.
Tabla 3. Contenido de fenoles totales, flavonoides, taninos condensados y antocianinas de hojas de cinco formas
silvestres de frijol común (Phaseolus vulgaris) de Durango, México.
Localities
Total phenolics1
Flavonoids2
Condensed
tannins3
Anthocyanins4
El Mezquital
550.08±18.18 a
212.68 ± 14.27 a
7.53 ± 0.34 b
7.02 ± 0.50 b
Súchil
276.85 ± 3.97 d
49.57 ± 3.49 d
5.51 ± 0.20 d
10.09 ± 1.76 a
Nombre de Dios
145.27 ± 13.78
e
48.45 ± 8.78 d
6.68 ± 0.15 c
3.89 ± 1.71 c
Canatlán
828.98 ± 22.52
a
94.17 ± 2.89 c
10.91 ± 0.14 a
5.76 ± 0.35 bc
Nuevo Ideal
398.45 ± 8.23 c
124.87 ± 3.06 b
5.20 ± 0.14 d
11.54 ± 2.32 a
Pr > F(Model)
<0.0001
<0.0001
<0.0001
0.186
Significant
Yes
Yes
Yes
No
1 Expressed in mg GAE / 100 g DW, 2 In mg QE / 100 g DW, 3 In mg EC / 100 g DW, 4 In mg C3OG / 100 g DW.
Different uppercase letters indicate significant differences between five populations (p ≤ 0.05) (Fisher’s test),
media ± SD (n = 3), DE (Dry extract), DW (Dry weight).
3.3 PCA and cluster analysis
Principal component analysis (Fig. 2A) showed that principal component 1 (PC1) explained 95.58
% of the total variance. The first three components explain 99.99 %. The variable with the greatest
contribution expressed by the PC1 was condensed tannins, with 95.58 %, PC2, flavonoids and total
phenols, with 3.06 % of the variance, and the PC3 was anthocyanins with 1.55 %. The dendrogram
of Fig. 2B indicates the grouping of the populations based on their similarity of contents of phenolic
compounds, which revealed two groups, one for seeds and the other for leaves, within the group of
leaves a greater similarity between the populations of Canatlán and Nuevo Ideal and within the
group of seeds a greater similarity between the populations of Canatlán and El Mezquital and
Nombre de Dios and Nuevo Ideal.
It has been reported that the pigments in seeds are attributed to phenolic compounds. A similar
study conducted by Armendáriz, et al. (2019) informed a phenol concentration ranges from 19.75 mg
to 221.48 mg gallic acid equivalents / g extract, in bean varieties grown with traditional methods in
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Oaxaca, Mexico. However, the values obtained in our study range from 133.39 to 269.90 mg gallic
acid equivalents/ g seed extracts.
The correlation between seed color and phenolic content may be crucial for understanding how these
characteristics affect seed quality and potentially their survival and adaptation in different
environmental conditions (Arteaga, 2021). Additionally, it is relevant to consider the role that these
compounds may have not only in the physical properties of seeds and leaves but also in antioxidant
properties and, therefore, in plant health (Colina, 2016; Bedoya & Maldonado, 2022; Sahu et al., 2022).
Figure 2. A) Results of a principal components analysis and B) dendrogram resulting from the comparison of
the different phenolic contents of seeds and leaves of five wild populations of common bean (Phaseolus
vulgaris) from Durango, Mexico.
Figura 2. A) Resultados del análisis de componentes principales y B) dendrograma resultante de la comparación
de los diferentes contenidos fenólicos de semillas y hojas de cinco poblaciones silvestres de frijol común
(Phaseolus vulgaris) de Durango, México.
3.4 HPLC-DAD
The results of the HPLC-DAD analysis of the leaves and seeds of wild forms of common beans
revealed this species is a rich source of phenolic compounds (Fig. 3). A total of 37 compounds were
identified from wild common beans with this technique. Flavonols and phenolic acid were the major
phenolic compounds in seeds (Table 4). The results are similar with those published by Reyes et al.
(2014) and Capistrán et al. (2019), who also found 27 phenolics in the leaves of P. vulgaris, the major
compounds were flavonols and phenolic acid.
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Figure 3. HPLC-DAD UV chromatograms of the leaves (A, registered at 265 nm) and seeds (B, registered at
340 nm) of wild forms of common bean from different regions of provenance of Durango, Mexico.
Figura 3. Cromatogramas UV HPLC-DAD de las hojas (A, registradas a 265 nm) y semillas (B, registradas a
340 nm) de formas silvestres de frijol común de diferentes regiones de procedencia de Durango, México.
The appearance of several absorption peaks, this fine structure reflects not only the different
conformations such systems may assume, but also electronic transitions between the different
vibrational energy levels possible for each electronic state. In the Table 4 are displayed the retention
time and spectral data for each compound present in leaves and seeds. There are compounds that
were observed both in leaves and seeds, but of the total of compounds eight of them were only found
in seeds at the retention times of 36.6 min (Aromatic acid), 33.29 min (Flavonols), 35.15 min
(Flavonols), 46.37 min (Dihydroflavonoids), 48.38 min (Flavonols), 49.35 min (Dihydroflavonoids),
55.36 min (Flavonols) and 57.35 min (Flavonols) and three were only found in leaves, at the retention
times 34.58 min (Dihydroflavonols), 36.46 min (Flavonols) and 40.33 min (Flavonols).
Chromatograms and UV spectra of some of the major compounds are shown in Fig. 4.
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Table 4. Phenolic compounds found in leaves and seeds of wild forms of common bean from Durango, Mexico.
Tabla 4. Compuestos fenólicos encontrados en hojas y semillas de formas silvestres de frijol común de Durango,
México.
Number of
compund
Retention time
(min)
ƛmax
Type of phenolic
compound
Type of sample
1
21.663 ± 0.33
325
Aromatic acid
seeds
2
22.32 ± 0.00
240sh-296sh-325
Aromatic acid
seeds
3
22.82 ± 0.00
245sh-296sh-323
Aromatic acid
seeds
4
23.44 ± 0.39
239sh-296sh.323
Aromatic acid
seeds
5
24.54 ± 0.50
325
Aromatic acid
seeds
6
26.39 ± 0.43
274
Aromatic acid
Leaves and seeds
7
27.58 ± 0.33
293sh-308
Aromatic acid
Leaves and seeds
8
28.67 ± 0.33
293sh-308
Aromatic acid
Leaves and seeds
9
31.55 ± 0.33
289-324sh
Dihydroflavonoids
Leaves and seeds
10
31.60 ± 0.00
293sh-308
Aromatic acid
Seeds
11
32.48 ± 0.31
239sh-293sh-323
Aromatic acid
Leaves and seeds
12
33.29 ± 0.00
249-263sh-298sh344
Flavonols
Seeds
13
33.45 ± 0.23
239sh-293sh-323
Aromatic acid
Leaves and seeds
14
34.58 ± 0.22
233sh-274-320sh
Dihydroflavonoids
Leaves
15
34.60 ± 0.00
288-334sh
Dihydroflavonoids
Leaves and seeds
16
35.14 ± 0.09
254-263sh-294sh348
Flavonols
Seeds
17
35.69 ± 0.12
293sh-308
Aromatic acid
Leaves and seeds
18
36.4 ± 0.00
274-329
Flavones
Leaves and seeds
19
36.46 ± 0.18
251-266sh-346
Flavonols
Leaves
20
37.00 ± 0.00
254sh-268-290sh-350
Flavones
Leaves and seeds
21
37.52 ± 0.27
255-266sh-294sh355
Flavonols
Leaves and seeds
22
38.42 ± 0.26
262-292sh-315sh-342
Flavonols
Leaves and seeds
23
38.60 ± 0.00
253-266sh-294sh352
Flavonols
Leaves and seeds
24
39.30 ± 0.00
252-262sh-
290sh318sh-352
Flavonols
Leaves and seeds
25
39.50 ± 0.30
238sh- 270-295sh330
Flavonols
Leaves and seeds
26
40.10 ± 0.00
262-292sh-315sh-342
Flavonols
Leaves and seeds
27
40.33 ± 0.27
254-263sh-294sh348
Flavonols
Leaves
13
Pérez-Salinas et.al
28
41.70 ± 0.31
276
Aromatic acid
Leaves and seeds
29
41.80 ± 0.00
255-295sh-372
Flavonols
Leaves and seeds
30
42.21 ± 0.10
233sh-273-320sh
Aromatic acid
Leaves and seeds
31
42.70 ± 0.17
265-290sh-320sh-346
Flavonols
Leaves and seeds
32
43.62 ± 0.16
255-263sh-297sh-357
Flavonols
Leaves and seeds
33
46.37 ± 0.05
289-326sh
Dihydroflavonoids
Seeds
34
48.38 ± 0.25
240-292sh-330
Flavonols
Seeds
35
49.35 ± 0.12
289-326sh
Dihydroflavonoids
Seeds
36
55.36 ± 0.28
248-262sh-318sh-346
Flavonols
Seeds
37
57.35 ± 0.64
230sh-248-
312sh327sh-360
Flavonols
Seeds
Figure 4. HPLC-DAD UV spectra (registered between 200 and 400 nm) of some phenolic compounds found in
the methanol extract of leaves and seeds of wild forms of common bean from Durango, Mexico.
Figura 4. Espectros UV HPLC-DAD (registrados entre 200 y 400 nm) de algunos compuestos fenólicos
encontrados en el extracto metanólico de hojas y semillas de formas silvestres de frijol común de
Durango, México.
Phenolic compounds are an important group of secondary metabolites found in various plants,
including the wild forms of common bean (Phaseolus vulgaris) (Espinoza et al., 2016; Capistrán, 2019).
In the methanolic extract of leaves and seeds of these varieties, several phenolic compounds are
usually identified, which may include simple phenolics, flavonoids, tannins, and phenolic acids. This
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TECNOCIENCIA CHIHUAHUA, Vol. XIX (1): e1691 (enero-abril. 2025)
study primarily found phenolic acids, known for their antioxidant properties (García-Díaz, 2016;
Claros, 2021; Pérez et al., 2020), and flavonols, such as quercetin and kaempferol, which have anti-
inflammatory and antioxidant effects (Capistrán, 2019).
4. Conclusions
The results provide valuable insight into the variability in phytochemical content among different
forms of wild beans, which could have significant implications for their use in breeding programs.
The differences in anthocyanin, flavonoid, total phenolic, and tannin content between seeds and
leaves suggest that each part of the plant may have different functions and benefits in terms of health
and adaptation. The finding that the seeds from Nuevo Ideal exhibit the highest contents of total
phenolic compounds (TPC), flavonoids (FC), condensed tannins (TCT), and anthocyanins (AC) is
particularly relevant, as it indicates that this variety may have greater nutraceutical potential. On the
other hand, the fact that Canatlán shows higher contents of TPC and TCT in the leaf tissue suggests
that this population could be explored for phytochemical applications in health products or as
antioxidant agents.
The variability in phenolic compound levels in populations growing at different altitudes also opens
the door for future studies on how environmental conditions affect the biosynthesis of these
metabolites. This could lead to a better understanding of plant adaptations to their environments
and how the cultivation of wild beans can be optimized to maximize their phytochemical content. It
would be interesting to consider the possibility of correlating this data with agroecological factors,
such as water availability, sunlight, and soil nutrients, to have a more comprehensive overview.
Additionally, investigating how these compounds affect resistance to pests and diseases could be
key to ensuring the sustainability of wild bean populations in the future.
Acknowledgements
To CONAHCYT and the National Polytechnic Institute and for their support in carrying out
this work.
Conflict of interest
This article was prepared for academic and scientific purposes only, noting that there is no
conflict of interest in the publication of these results.
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