Next Article in Journal
Genetic and Genomic Resources for Soybean Breeding Research
Next Article in Special Issue
Chemical Profile, Antibacterial and Antioxidant Potential of Zingiber officinale Roscoe and Elettaria cardamomum (L.) Maton Essential Oils and Extracts
Previous Article in Journal
Impact of Climate Change on Phenology of Two Heat-Resistant Wheat Varieties and Future Adaptations
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Plants
Volume 11
Issue 9
10.3390/plants11091179
plants-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Antioxidant Capacity and Antiplatelet Activity of Aqueous Extracts of Common Bean (Phaseolus vulgaris L.) Obtained with Microwave and Ultrasound Assisted Extraction

by
Lyanne Rodríguez
1,†,
Andrea Plaza
1,†,
Diego Méndez
1,
Basilio Carrasco
1,
Francisca Tellería
2,
Iván Palomo
2,* and
Eduardo Fuentes
2,*
1
Centro de Estudios en Alimentos Procesados (CEAP), CONICYT Programa Regional, Gore Maule R0912001, Casilla 1007, Talca 3480094, Chile
2
Thrombosis Research Center, Medical Technology School, Department of Clinical Biochemistry and Immunohaematology, Faculty of Health Sciences, Universidad de Talca, Talca 3480094, Chile
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Plants 2022, 11(9), 1179; https://doi.org/10.3390/plants11091179
Submission received: 17 March 2022 / Revised: 11 April 2022 / Accepted: 20 April 2022 / Published: 27 April 2022
(This article belongs to the Special Issue Phytochemical and Antioxidant Properties of Plants—Health Care, Food Production and Green Agriculture)
Browse Figures
Versions Notes

Abstract

:
Phaseolus vulgaris L. has beneficial effects on several chronic non-communicable diseases (e.g., cardiovascular diseases) related to oxidative stress. This redox state may influence platelet activation and aggregation; which is crucial in thrombus formation. In this work, the antiplatelet and antioxidant potential of aqueous extracts obtained by green processes, microwave-assisted extraction and ultrasound-assisted extraction, from 25 landraces of common beans were investigated. Phenol content and antioxidant potential were determined using the Folin-Ciocalteu method, total monomeric anthocyanin and ORAC assay, respectively. The antiplatelet potential of the extracts was explored by turbidimetry. Microwave extraction showed higher phenol content and antioxidant activity in most extracts. Soja landrace extract obtained by microwave-assisted extraction showed higher phenol content and antioxidant activity (893.45 ± 87.30 mg GAE/g and 35,642.85 ± 2588.88 ORAC μmolTE/g, respectively). Although most of the extracts obtained by microwave-assisted extraction showed antiplatelet activity, the extract of Hallado Aleman landrace obtained by ultrasound-assisted extraction (IC50 = 0.152 ± 0.018 mg/mL) had the highest antiplatelet potential. The extraction method, MAE and UAE, influences the biological potential of the beans, specifically the antiplatelet activity and antioxidant activity. The functional value of this legume for direct consumption by the population was evidenced, as well as its inclusion in food formulations.

Graphical Abstract

1. Introduction

Cardiovascular diseases (CVD) account for most chronic non-communicable diseases deaths, (17.9 million people annually), followed by cancers (9.0 million), respiratory diseases (3.9 million), and diabetes (1.6 million) [1]. Oxidative stress is a factor of great importance in the progression of these chronic diseases [2,3]. In the case of CVD, oxidative stress and platelet activation are often closely related. Thus the increase in Reactive oxygen species (ROS) in the circulation exposes the platelets to an activating medium, promoting the change of platelet phenotype to a pro-adhesive one and aggregation, which in turn leads to thromboembolic propensity [4,5].
Given the high prevalence and pathophysiology of CVD; exists an increase in demand for food sources that contribute to its prevention [6,7]. In recent years, the study of bioactive compounds derived from fruits and vegetables capable of generating antiplatelet activity, as well as their mechanism of action, has been increasing. In this way, it has been identified that mango (Mangifera indica), maqui (Aristotelia chilensis), guava (Psidium guajava), turmeric (Curcuma longa), and tomato, among others, have antiplatelet potential, mainly associated with a high content of phenolic compounds [8,9,10].
The common bean (Phaseolus vulgaris L.) is a major grain legume consumed worldwide and, depending on the type of cultivar, are mainly composed of protein (9 to 50 g/100 g), carbohydrates (20 to 75 g/100 g), lipids (1.5 to 25 g/100 g), dietary fiber (14 to 19 g/100 g) [11] as well as been rich in bioactive compounds [12].
Several studies show the broad chemical profile of common beans, where phenolic acids stand out (p-coumaric acid, synaptic acid, ferulic acid, feruloyl aldaric acid, p- hydroxybenzoic acid, protocatechuic acid, vanillic acid and syringic acid), isoflavonoids (genistein, daidzein, glycitein, formononetin), flavanones (naringenin and hesperetin), anthocyanins (delphinidin, petunidin, malvidin and cyanidin, pelargonidin), sugars, fatty acids, and tocopherols [13,14], among many other compounds that have been reported by our group previously [13]. The content of bioactive compounds varies according to the landrace [14,15,16] and extraction technique [17,18]. Table S1 shows the content of bioactive compounds for different landraces and the extraction technique used.
Some of the problems presented by conventional extraction technologies are long extraction times, the use of expensive and high-purity solvents, evaporation of large amounts of solvent due to the high temperatures used, low extraction selectivity, and thermal decomposition of thermolabile compounds [19,20]. To overcome these problems, unconventional techniques emerged, also called green or clean [18,20,21]. Green extraction methods have been developed using modern technologies, in which less or no organic solvents are used to minimize health and environmental impacts and maximize the yield of desired polyphenols through selective extraction. A definition of green extraction of natural products is “Green extraction is based on the discovery and design of extraction process which will reduce energy consumption, allows use of alternative solvents and renewable natural products, and ensure a safe and high-quality extract/product” [22,23].
The most widely used green processes are ultrasound-assisted extraction (UAE), supercritical fluids (SCFs), microwave-assisted extraction (MAE), and pressurized liquid extraction (PLE) [24,25].
Generally speaking, microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE) are promising techniques when compared to the conventional ones, because they are simple and accessible methods that give high yields in a shorter time with minimal solvent and they can be used with different plant materials, including common beans [18].
MAE is carried out using non-ionizing microwave radiation (100–900 W) to the hydrated sample. The sample is subjected to the irradiation of microwave waves causing the movement of the molecules by migration of ions and dipoles that contribute to the increase in temperature, inducing cell wall destruction, and then facilitating the diffusion of compounds from the matrix to the solvent [26]. Sutivisedsak et al. showed that MAE is an effective method for the extraction of phytochemical phenolics in 8 types of North American beans. The content of total phenols using MAE and water as solvent at 100 °C was two to three times higher than the values obtained by conventional extraction [16].
UAE offers substantial advantages over conventional methods compared to traditional extraction methods since the propagation of ultrasound waves (over 20 kHz) creates a breakdown of cellular material by the cavitation phenomenon, which generates a better penetration of solvents in cellular materials and reduces the duration of the process [27,28]. Indeed, ultrasound-assisted extraction technology can potentially improve the extraction of components such as polyphenols, anthocyanins, aromatics, polysaccharides, oils, and functional compounds when is used as a pretreatment in a processing unit [29]. Yang et al., 2019, revealed that UAE exhibits a higher extraction rate of antioxidants from common bean, showing an extraction efficiency seven times higher than conventional solvent extraction. Some of the compounds detected in this study were Protocatechuic acid, catechin, chlorogenic acid, epicatechin, ferulic acid, coumarin, rutin, myricetin, cinnamic acid, and genistein [30].
In this work we try to value a food widely consumed by the population in Chile and the rest of the world, due to its high nutritional value, rich in fiber, protein, starch, carbohydrates, phenols and low in fat. We consider that the antiplatelet potential of common bean has been little studied, there is only the background of Rodriguez-Arzua et al., 2018, studied landraces of common bean in different stages of growth (green beans, with green pods and green grain; fresh beans, shelled). They evaluated aqueous and methanolic extracts, obtained by maceration and sonication, respectively. Showed that aqueous and methanolic extracts of fresh beans, as well as green beans, showed a similar antiplatelet effect against the agonists ADP and arachidonic acid [31].
In this work, 25 Chilean landraces, very little evaluated to date, were studied. The extraction conditions, ultrasound-assisted extraction technique and microwave-assisted extraction technique were investigated to enhance the protective functionality of this legume. Additionally, we tried to show if there was any relationship between the antioxidant activity evaluated by the ORAC assay and the antiplatelet activity. This allowed us to make a comparison of the extraction methodologies used.

2. Results

2.1. Extraction Yields of Extracts P. vulgaris L.

The extraction yields in terms of the soluble solid compounds extracted by the microwave and ultrasound-assisted solid-liquid extractions are shown in Table 1.
The soluble solids of the different landraces of Chilean common bean ranged between 0.87 ± 0.06 and 2.27 ± 0.06 Brix. These results indicate the amount of non-water soluble compounds in these extracts. The microwave-assisted extraction showed a higher content of total solids except for the landraces of Blanco Español, Palo, Cabrita, Torcaza, and Cimarron, where the ultrasound extraction showed a higher content of total solids than microwave-assisted extraction. The Cimarron landrace had higher total solids content than the rest of the bean landrace, both for microwave-assisted extraction and for ultrasound-assisted extraction. The Cimarron, Tortola, and Soja extracts obtained by microwave-assisted extraction had twice the total solids content compared to the Bombero extract, this showed the lowest total solids content of 0.87 ± 0.06 Brix.

2.2. Total Phenolic Content of Extracts P. vulgaris L.

Previous studies (data not shown) carried out using extraction of compounds with antiplatelet activity using magnetic agitation and orbital shaker showed that the extracts obtained had a lower efficiency in inhibiting platelet aggregation than those extracts obtained by MAE and UAE. It is for this reason that the results obtained with the extracts prepared by MAE and UAE are shown in this work.
Of the 25 landraces of beans, the extracts made by microwave-assisted extraction showed the highest content of phenols in general, highlighting the landraces of Soja and Peumo (893.45 ± 87.30, 866.17 ± 30.28 mg GAE/g, respectively), with the highest contents of total phenols (Table 1). These extracts showed 4 times higher content of total phenols than the microwave-assisted Cisne extract, which showed a lower content of phenols, 220.08 ± 11.98 mg GAE/g shows that the rest of the extracts. The lowest phenol content was obtained in the landraces assisted by ultrasound, except for the landraces of Mantequilla, Lunatus, Cabrita, Pallar Manchado, Blanco Español, and Cisne, where microwave-assisted extraction showed a lower content of total phenols.

2.3. Antioxidant Activity of Extracts P. vulgaris L.

In the antioxidant activity using the ORAC test, the extracts made by microwave-assisted extraction also stood out Frutilla (Table 1). The Soja extract obtained by MAE had the highest antioxidant capacity, 35,642.85 ± 2588.88 μmolTE/g, followed by the Peumo and Araucano. These landraces also had the highest total phenol content as we saw earlier. The Lunatus extract using ultrasound-assisted extraction showed the lowest antioxidant capacity, 797.13 ± 158.30 μmolTE/g. Additionally, the extract of Coscorron assisted by ultrasound showed negative ORAC values, the value was below the limit of quantification of the calibration curve. This extract also showed a low content of total phenols, which can be shown in Table 1.

2.4. Cytotoxicity of P. vulgaris L. Extracts on Platelets

Figure 1 shows the effect of MAE and UAE extracts on LDH release from platelets (cytotoxicity). Values are expressed as mean percentage and SD of 6 independent tests. It is observed 50 extracts (MAE and UAE) do not produce cytotoxicity in washed platelets when compared with the control (vehicle).

2.5. Platelet Antiaggregant Activity of Extracts P. vulgaris L.

The antiplatelet screening was carried out with platelet-rich plasma from healthy volunteers where the antiplatelet activity of the 25 landraces of common bean produced by microwave and ultrasound-assisted extraction was evaluated (Table S2). The study of antiplatelet activity showed that bean extracts have greater potential when platelet aggregation is stimulated by TRAP-6 10 µM than by ADP 4 µM. In the case of the latter agonist, the extracts that had the highest percentage of platelet inhibition, around 20%, were the Arauco and Cimarron extracts. While, when platelet aggregation was induced with TRAP-6, significant percentages of platelet aggregation inhibition were observed compared to the control. The extracts made by microwave-assisted extraction showed greater antiplatelet potential than the extracts made by ultrasound-assisted extraction, except for the landraces of Bombero, Cimarron, Hallado Aleman, Lunatus, Palo, Pallar Manchado, and Pallar Morado. These extracts obtained by ultrasound-assisted extraction showed high antiplatelet potential when aggregation was stimulated by TRAP-6 (Table 1).
The most active extracts (percentage of platelet inhibition above 50%) obtained by MAE and UAE, from the landraces of common bean, were selected to evaluate the concentration-dependent effect on antiplatelet activity (Figure 2).
24 landraces of beans were studied, of which only 9 extracts were obtained by UAE, the rest by MAE. Although the extracts that showed the greatest inhibition of platelet aggregation were Hallado Aleman (UAE) and Pallar Manchado (UAE), IC50 = 0.152 ± 0.018, and 0.165 ± 0.015 mg/mL, respectively.

2.6. Total Monomeric Anthocyanin Content of Extracts P. vulgaris L.

Total monomeric anthocyanin contents assays were done to the extracts with more platelet inhibition, these results are show in Table 2.
Based on the results of the total monomeric anthocyanin content analysis performed on bean extracts with the highest platelet inhibition showed in Table 2, it is possible to indicate that the extract with the highest total monomeric anthocyanin content is Pallar Morado followed for Ganso both extracts obtained with MAE extraction process. This higher content of total monomeric anthocyanins could be explained by the presence of spots on the skin of the bean.

3. Discussion

In this investigation, extractions by green processes at laboratory scale, MAE and UAE were used. The use of these unconventional techniques provides shorter extraction time, better extraction selectivity, and less thermal decomposition of thermolabile compounds [19,20]. In this study the power used in MAE is low and in UAE the Erlenmeyer flask with the mixture bean/water is submerged in a container with water, which prevents the working temperature from increasing above 50 °C in both methodologies. This would allow avoiding the degradation of thermolabile compounds. Furthermore, they have reduced energy use (mainly due to the speed of the techniques, which implies less energy use), in addition to being technologies that offer high extraction performance, as well as scalable to the industry [18,20,21]. Industrial applications of these processes include the extraction of many natural compounds, such as vitamins, aromas, natural pigments, or essential oils [29,32].
However, there is some evidence that suggest that prolonged exposure to microwaves of high-power levels during MAE may leads to degradation of phenolics compounds (due to their ability to react between them and form smaller or larger compounds as well as form new ones during heating) and the formation of cavitation bubbles during UAE can generates hydroxyl radicals, which can also result in the deterioration of phenolics compounds [18,33,34]. In the methods used, there was a relationship between the selected parameters (mainly time, temperature and solvent), in order to avoid the aforementioned processes.
The results obtained here showed that MAE had better results than UAE. The extracts obtained by the latter generally showed a lower content of total phenols and a lower antioxidant activity. In recent years MAE factors such as potency, solvent, solvent ratio, time, and temperature have been optimized to achieve a high yield of phenolic compounds [18,35].
The MAE process can be relatively selective in the extraction of certain compounds or groups of compounds since dielectric heating depends on the frequency and power of the microwaves [36]. In our study, it was shown that the extraction of phenolic compounds by this technique was carried out efficiently. Most of the common bean landraces studied showed higher ORAC values, highlighting the Soja, Peumo, and Araucano extracts these extracts also had a high content of phenolic compounds. Soja extract had the highest content of total phenols and also the highest antioxidant activity according to the ORAC test.
The results correlated with what was previously reported by Kumar et al., 2019, who reported that the extraction of anthocyanins and phenolic compounds from the seed coat of black Soja bean using MAE produced a maximum of total anthocyanins, 5094.9 mg/L. However, the conventional extraction yield was 1246.89 ± 68.45 mg/L [18,35]. Other authors have revealed that the MAE technique combined with ethanol as a solvent shows a higher yield and extraction quality of green coffee oil [37]. Microwave energy provided localized heating in plant cells, and its driving force could disrupt the plant matrix [38]. It has also been shown that extracts made from cocoa bean shell waste using optimal MAE conditions showed better results than conventional extraction [39]. Other authors have seen that water as a solvent and ultrasound extraction are effective for the extraction of total phenol content (TPC), total flavonoid content (TFC), and antioxidant activity of Mucuna macrocarpa (MM) beans [40].
Various assays have been frequently used to estimate antioxidant capacities in fresh fruits and vegetables and their products and foods for clinical studies [41]. The ORAC assay is more relevant because it uses a biologically relevant radical source [42]. Xu et al., 2007 investigated how solvents affect the phenol content and antioxidant capacity of 8 kinds of legumes. The results showed that the polarity of the solvent has significant effects on the total phenolic content, as well as on the rest of the extracted components. The 70% ethanol extracts exhibited the highest ORAC value for all selected legumes [43]. The seed coats of red sword bean (Canavalia gladiate (Jacq.) DC.) are rich in antioxidant polyphenols. Under optimal conditions, the antioxidant activity of the extract of this seed obtained by UAE was 755.98 ± 10.23 μmol Trolox/g dry weight, much higher than maceration (558.77 ± 14.42 μmol Trolox/g) or the Soxhlet extraction (479.81 ± 12.75 μmol Trolox/g) [44]. The bean landraces we studied showed higher antioxidant activity than the red sword bean seed coat except for the Coscorrón landrace UAE which could not be detected. The rest of the extracts had higher antioxidant activity according to the ORAC assay.
Except 7 landraces of common bean the rest of the extracts obtained by MAE showed greater antiplatelet potential, stimulated fundamentally with TRAP-6. Extracts of Hallado Aleman (UAE), Pallar Manchado (UAE), and Torcaza (MAE) stood out with higher antiplatelet activity, 0.152 ± 0.018, 0.165 ± 0.015, and 0.172 ± 0.049 mg/mL, respectively. This activity did not correspond to the higher content of phenols or higher antioxidant activity. Hallado Aleman extract had about 1.5 times more phenol content than the Cisne MAE extract (extract with lower content of phenolic compounds). On the other hand, its antioxidant capacity according to the ORAC assay was 14 times higher than the Lunatus extract (lower antioxidant activity by ORAC assay).
There have been few reports showing antiplatelet activity for this legume. Rodriguez et al. [45], showed that the content of total phenols, the content of total flavonoids, the content of total monomeric anthocyanins, and antioxidant capacity in 255 lines of common bean grown under the same environmental conditions have a wide range of variability, with differences always above of an order of magnitude. They showed that bean extracts significantly reduced platelet aggregation induced by adenosine 5′-diphosphate and arachidonic acid.
A large number of metabolites with antiplatelet potential have been identified in common beans (Table 3). An HPLC-MS study of the Black Jamapa bean seed coat revealed the presence of flavonols, anthocyanins, flavanol monomers, and heterogeneous flavanol oligomers. This work reported for the first time in this legume the presence of myricetin glycoside and proanthocyanidin oligomers containing (epi)gallocatechin [46]. Yang et al., 2019, also showed the presence of myricetin, these authors revealed that UAE exhibits a high extraction rate of antioxidants from common bean, showing an extraction efficiency seven times higher than conventional solvent extraction. Some of the compounds detected by this technique were myricetin, cinnamic acid and genistein [30]. Myricetin has been reported to reduce the ability of platelets to spread over collagen and form thrombo in vitro [47]. Genistein suppresses in vitro human platelet aggregation, serotonin secretion, and collagen- and TxA2-induced protein tyrosine phosphorylation. It slightly attenuates thrombin-induced protein tyrosine phosphorylation (100 μg/mL) [47]. Studies suggest that phenolic acid can inhibit platelet adhesion and aggregation [48,49,50,51,52], LDL oxidation [50], protects the heart against oxidative stress [53,54], inhibits platelet inflammatory mediators [50], Inhibit P-selectin expression [49,53] and increases cAMP levels (6,8,9), among others. Along the same lines, flavonoids have also shown the ability to inhibit platelet aggregation induced by various agonists [55,56,57,58], protect against cerebral ischemia and reperfusion injury in rats, enhances cerebrovascular angiogenesis in human umbilical vein endothelial cells [59], inhibit PI3 production [60,61], increases cAMP levels [56] and inhibits levels of inflammatory markers [62]. Anthocyanins have also shown effects similar to those mentioned before [63,64] and, in addition, inhibit P-selectin expression [63,64,65], inhibit the secretion of alpha and dense granules, inhibit PI3K/Akt activation, eNOS phosphorylation and cGMP production [65].
The results of Pearson’s correlation (Figure S1), showed that common bean extracts relationship with phenol content and antioxidant activity (r = 0.68). On the other hand, neither the antioxidant activity nor the phenol content showed a relationship with the antiplatelet activity evaluated in the extracts (r = 0.0037 and r = −0.096, respectively). The antiplatelet activity of numerous fruits and vegetables has been reported to be associated with multiple mechanisms of action [68]. ROS generation has been described as a critical step required for platelet activation, as these species represent important secondary messengers in signal transduction cascades, which may be required for the propagation and activation of platelet aggregation [69]. Sometimes the inhibition of platelet aggregation has been related to the presence of antioxidants. On the other hand, platelets are regulators of multiple processes, with the recruitment of inflammatory cells to sites of injury, release of inflammatory mediators, and regulation of endothelial function [70]. There are many inhibition pathways that have been described for natural products far from a direct effect on ROS, we can mention inhibition of platelet phospholipase C (rutaecarpine, resveratrol and rutin), increased levels of cAMP (allicin, disulfiram, quercetin and guanosine), TXA2 and thrombin receptor antagonists (equol and apigenin), and inositol monophosphate inhibition (marchantinquinone) [71].
It has been shown that the difference between the antiplatelet potential can be attributed to the individual action of each secondary metabolite and can be enhanced from their interactions through a supposed synergy or cooperativity between the metabolites [8]. However, it has been shown that there are many factors that influence the composition of an extract and therefore its biological potential, among these variables we can mention genotype, solvent, extraction method, environment, processing, storage and harvest stage [8,72]. A study carried out on the berries showed that the antiplatelet activity varies according to the genotype, the solvent used and the organ of the plant, as well as the state of maturity of the fruit. Rodriguez et al., 2018. revealed that the immature fruit had a high antiplatelet potential, which was related to its high phenol content and lower anthocyanin content, unlike the mature fruit, which was richer in anthocyanins and has been described as having a high antioxidant potential [72,73]. These results are evidence that antioxidant activity is sometimes not related to a high antiplatelet potential. The same was observed in this study, where it was shown that extracts with higher phenol content and higher ORAC value (antioxidant capacity) did not have greater antiplatelet potential. In other works, we will study the chemical profile of the extracts with the highest antioxidant and antiplatelet activity, in order to show the differences in the chemical composition of each exact, which should be related to the findings of antiplatelet inhibition found for the extracts of common bean.

4. Materials and Methods

4.1. Plant Material

Samples of the 24 different Chilean landraces Phaseolus vulgaris L. were obtained from small producers in the south-central macrozone of Chile (Table 4), all bean samples corresponded to landraces from the year 2021, Bean cultivars are mainly composed of protein, dietary fiber, minerals, and vitamins [74].
Once the samples were obtained, they were ground using a domestic juicer, and subsequently sieved with a 450-micron sieve, mesh 40. This grinding was carried out to reduce the size of the particles, to increase the effective contact area between the particle and water, used as extraction solvent as is described for Panja [23], and for Llompart and coworkers [75]. The above procedure then allows increasing the extraction yields of target compounds.

4.2. Preparation of Extracts

To obtain the extracts, about 1 g of each of the 25 previously ground bean samples was used and put in contact with 40 milliliters of distilled water in an Erlenmeyer flask. Thus, extraction assays were developed using a ratio of beans/water of 1/40, for one hour. This relationship was selected to not saturate the extraction solvent, and thus increase the extraction yields. Microwave-assisted solid-liquid extractions were carried out under the next operational variables:
  • Ultrasound-assisted extraction (UAE): An ultrasound bar (QSonica Q125, Newton, CT, USA) was used, at 50% amplitude (10 kHz) with a sonication time of 60 min, to prevent the temperature from rising too high, the Erlenmeyer flask with the mixture of beans and distilled water was placed inside of an ice bath.
  • Microwave-assisted extraction (MAE): A 1200 W domestic microwave (WMW606ADWC Whirlpool, Shanghai, China) was used. To carry out the microwave-assisted extraction, the bean mixtures with distilled water in an Erlenmeyer flask, were placed inside them in a 1/40 ratio. The minimum power of 1 was fixed, which is equivalent to 120 W. The extraction time was 60 min.
The samples obtained in the microwave and ultrasound-assisted extraction processes were centrifuged 50 mL conical tubes in a centrifuge (K2015R Centurion Scientific, Manchester, UK) at 3500 rpm for 15 min at 20 °C. Subsequently, the supernatant was separated with Corning Falcon cell filters of 100, 70, and 40 µm respectively, to obtain the supernatant, which was frozen at −86 °C for 48 h, then freeze-dried (Operon, FDU 7024, Gimpo, Korea) for 20 h with a cold trap (−70 °C) and stored until analysis.

4.3. Total Soluble Solids and Total Phenolics

For the determination of soluble solids, a digital refractometer (Hanna Instruments HI 96801, Woonsocket, RI, USA) was used. On the calibrated refractometer and at room temperature (20 °C), 0.5 mL of filtered supernatant was deposited, and the result was read and expressed as °Brix, which is equivalent to the weight/weight percentage. The reading procedure was carried out in triplicate.
On the other hand, total phenol content was determined by the Folin-Ciocalteu method. This method quantifies the reducing power of phenolic compounds on the Folin-Ciocalteu reagent, through the formation of a blue complex that is read at 750 nm [76]. The methodology used is based on OIV-MA-AS2-10, Compendium of International Methods of Analysis—OIV Folin-Ciocalteu Index, 2019 edition. Briefly, a calibration curve was made with different concentrations from a primary solution of 5 g/L of gallic acid. For the reading of the samples, the following procedure was followed. In a 10 mL flask, the reagents were added in the following order: 250 µL of sample, standard or water in the case of the blank; 5 mL of MilliQ water; 500 µL of Folin’s reagent; 2 mL of sodium carbonate p/v solution. It was made up to volume with MiliQ water, stirred and subsequently stored for 30 min in the dark at room temperature. Finally, absorbance was measured in a spectrophotometer (Spectro UV-11, MCR, Shanghai, China) at 750 nm.

4.4. Total Monomeric Anthocyanin Content

Determination of total monomeric anthocyanin content was determined according to the official method of the AOAC [77]. This method is based on the measurement of the color change of monomeric anthocyanins pigments, which change color reversibly with a change in pH; the colored form of oxonium exists at pH 1.0, and the colorless hemiketal form predominates at pH 4.5. The difference in the absorbance of the pigments at 520 nm is proportional to the pigment concentration. The results are expressed in a cyanidin-3-glucoside base. It should be noted that anthocyanins degraded in polymeric forms are not included in the measurements because they absorb at pH 4.5 as well as pH 1.0, determine the appropriate dilution factor by diluting the sample with pH 1 buffer, until absorbance at 520 is within the linear range of the spectrophotometer.
To carry out this measurement, a pH 1 buffer of potassium chloride (0.025 M) and a pH 4.5 buffer of sodium acetate (0.4 M) were used. Two dilutions of the samples to be analyzed were prepared, one with the pH 1 buffer and the other using the pH 4.5 buffer, subsequently the absorbance at 520 and 700 nm was determined against a blank cell, which was filled with distilled water.
Total monomeric anthocyanins were calculated using the following equation:
T o t a l   m o n o m e r i c   A n t h o c y a n i n   ( c y a n i d i n − 3 − g l u c o s i d e   e q u i v a l e n t s ,   m g / L = A ∗   M W ∗ D F   x 10 3 Ɛ ∗ I
where A = (A520nm − A700nm) pH 1.0 − (A520nm − A700nm) pH 4.5; MW (Molecular Weight) = 449.2 g mol−1 for cyanidin-3-glucoside; DF = Dilution Factor; l = pathlength in cm; Ɛ = 26 900 molar extinction coefficient, in L × mol−1 cm−1, for cyanidin-3-glucoside; and 103 = factor for conversion from g to mg.

4.5. Antioxidant Activity (ORAC)

There are various methodologies to measure the scavenging capacity of free radicals, such as ORAC, ABTS, DPPH, DMPD, among others. ORAC (Oxygen Radical Absorbance Capacity) is the analytical method most implemented for antioxidant analysis due to its adaptability to the great need of radical species and conditions of various laboratories. In addition to being done under conditions like those physiological and with the superoxide anion which is a radical with high biological relevance [78].
For the analysis of antioxidant capacity by ORAC, the metholody described for Ou and coworkers [79] was used a fluorescence microplate reader with an excitation filter at 485 ± 20 nm and an emission filter at 538 ± 25 nm (Synergy HTX Multi-Mode Reader, Biotek, Santa Clara, CA, USA) was used. This instrument is capable of evaluating UV-Vis absorbance, fluorescence, and luminescence. Briefly, sodium fluorescein (0.015 mg mL−1), AAPH radical solution (120 mg mL−1), and Trolox standard solution (100 µM) were prepared with phosphate buffer (75 mM, pH 7). The operating conditions for the final reaction consisted of 50 μL of diluted extract, Trolox standard or phosphate buffer (blank), 50 μL of fluorescein, and 25 μL of AAPH incubated at 37 °C in the microplate reader. For the measurement, black plates with 96 wells were used allowing the analysis of three samples per plate. Gallic acid was used as a control. The fluorescence was recorded every 5 min over 60 min using the KC4 TM software (BioTek Instruments, Winooski, VT, USA), differences of areas under the fluorescence decay curve (AUC) between the blank and the sample over time were compared and the results were expressed as μM Trolox Equivalents

4.6. Preparation of Human Platelets

Whole blood was obtained by venous phlebotomy from healthy volunteer donors (7 days without NSAID use) who accepted informed consent, as previously described [80]. The study protocol was approved by the Scientific Ethics Committee from Universidad de Talca (Protocol N° 29/2021). Whole blood was centrifuged at 240× g to obtain platelet-rich plasma (PRP) as previously described [8]. Subsequently, the platelet count was adjusted to 300 × 109 platelet/L with platelet-poor plasma (PPP) obtained by centrifugation at 800× g.

4.7. Obtaining Washed Platelets

Whole blood was anticoagulated with acid-citrate-dextrose (ACD) (v/v ratio 4:1) and centrifuged for 12 min at 240× g to obtain platelet-rich plasma (PRP). The PRP was centrifuged for 8 min at 900× g in a microcentrifuge. The pellet (platelets) was resuspended in Buffer Tyrodes without calcium: ACD (v/v ratio 5:1) and centrifuged again for 8 min at 900× g. Finally, the platelets were resuspended in Buffer Tyrodes without calcium and adjusted to the desired count by reading the hematology counter (Mindray BC-3000 Plus, Shenzhen, China) [81].

4.8. Release of Lactate Dehydrogenase (LDH)

To assess the cytotoxicity, the LDH Cytotoxicity Assay Kit (Cayman Chemical, Ann Arbor, MI, USA) was used. Washed platelets (200 × 109 platelets/L) were incubated with the highest concentration of extracts tested (1 mg/mL) or vehicle for 10 min at 37 °C. Subsequently, the platelet solution was centrifuged at 900× g for 8 min in a microcentrifuge to obtain the cell supernatant, which was reacted with the kit’s working reagent. As a positive control of 100% cytotoxicity (maximum LDH release), a 10% Triton X-100 solution was used on washed platelets [82].

4.9. Platelet Aggregation

The antiplatelet activity of the beans obtained by green processes was performed by turbidimetry using a lumi-agregometer as previously described [9]. The PRP was incubated with the different extracts at a concentration of 1 mg/mL for 4 min. Platelet aggregation was induced with the agonists ADP 4 µM and TRAP-6 10 µM, these agonists act on different platelet receptors [80]. Additionally, PRP was incubated with PBS as a positive control of platelet aggregation. The percentage of platelet aggregation was obtained with the AGGRO/LINK software (Chrono-Log, Havertown, PA, EE. UU.). The results were expressed as inhibition of platelet aggregation. Once the antiplatelet activity was evaluated at 1 mg/mL, the concentration-dependent antiplatelet potential was studied. The extracts that inhibited platelet aggregation by more than 50% were selected and the concentration necessary to reduce platelet aggregation by 50% (IC50) was determined from the concentration curves of the extracts of common bean (0.025, 0.05, 0.10, 0.25, 0.50, 0.75, 1.00 mg/mL).

4.10. Statistical Analysis

Data were analyzed using Prism 8.0 software (GraphPad Inc., San Diego, CA, USA) and expressed as mean ± standard deviation (SD). Considering the small size of the experiments to determine the normality of the data, the Shapiro-Wilk test was used. For the analysis of the platelet aggregation results, a one-way analysis of variance (ANOVA) was used to detect if there were statistical differences between the groups and the Dunnet’s postdoc test to make a comparison with the control (vehicle) [80]. While Tuckey’s was used to compare the differences between various groups, 25 landraces [83]. The differences between groups were analyzed using a one-way analysis of variance (ANOVA) and Tukey’s posthoc test. The Pearson correlation coefficient was used to determine the differences between the conditions and variables studied. p values <0.05 were considered statistically significant (Figure S1) [8].

5. Conclusions

This study evidence extraction efficiency between the green processes, MAE and UAE, to obtain extracts of 25 landraces of common bean with antioxidant and antiplatelet properties. Our studies reveal that the antiplatelet potential of bean extracts is not related to the presence of antioxidants (ROS inhibition pathway). Other studies are needed to help us elucidate the eventual mechanism by which the most active bean extracts inhibit platelet aggregation considering the chemical profile of the extracts and the different pathways mentioned above.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/plants11091179/s1, Table S1. Content of bioactive compounds for different varieties and extraction technique. Table S2. Antiplatelet activity of 25 varieties of P. vulgaris L. Figure S1. Correlation between phenol content, antioxidant activity, and antiplatelet activity of extracts P. vulgaris L. References [84,85,86,87] are cited in the supplementary materials.

Author Contributions

L.R. and A.P.: conceptualization, conducted the research, formal analysis, writing original draft. D.M. and F.T.: methodology and formal analysis. B.C.: conceptualization. I.P. and E.F.: conceptualization, resources, conducted the research, writing original draft, formal analysis. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Centro de Estudios en Alimentos Procesados-CEAP, ANID, Programa Regional R20F0001, Talca, Chile.

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki, and approved by the Scientific Ethics Committee of University of Talca (protocol code 29-2021 and date of approval 29 September 2021).

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. World Health Organization. Noncommunicable Diseases; World Health Organization: Geneva, Switzerland, 2018. [Google Scholar]
  2. Dubois-Deruy, E.; Peugnet, V.; Turkieh, A.; Pinet, F. Oxidative stress in cardiovascular diseases. Antioxidants 2020, 9, 864. [Google Scholar] [CrossRef] [PubMed]
  3. Pizzino, G.; Irrera, N.; Cucinotta, M.; Pallio, G.; Mannino, F.; Arcoraci, V.; Squadrito, F.; Altavilla, D.; Bitto, A. Oxidative stress: Harms and benefits for human health. Oxidative Med. Cell. Longev. 2017, 2017, 8416763. [Google Scholar] [CrossRef] [PubMed]
  4. Masselli, E.; Pozzi, G.; Vaccarezza, M.; Mirandola, P.; Galli, D.; Vitale, M.; Carubbi, C.; Gobbi, G. ROS in platelet biology: Functional aspects and methodological insights. Int. J. Mol. Sci. 2020, 21, 4866. [Google Scholar] [CrossRef] [PubMed]
  5. Calvieri, C.; Tanzilli, G.; Bartimoccia, S.; Cangemi, R.; Arrivi, A.; Dominici, M.; Cammisotto, V.; Viceconte, N.; Mangieri, E.; Frati, G. Interplay between oxidative stress and platelet activation in coronary thrombus of STEMI patients. Antioxidant 2018, 7, 83. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  6. Yu, E.; Malik, V.S.; Hu, F.B. Cardiovascular disease prevention by diet modification: JACC health promotion series. J. Am. Coll. Cardiol. 2018, 72, 914–926. [Google Scholar] [CrossRef]
  7. Lutz, M.; Fuentes, E.; Ávila, F.; Alarcón, M.; Palomo, I. Roles of phenolic compounds in the reduction of risk factors of cardiovascular diseases. Molecules 2019, 24, 366. [Google Scholar] [CrossRef] [Green Version]
  8. Rodríguez-Pérez, L. Antiplatelet effect of Aristotelia chilensis (maqui) extracts through in vitro studies. PLoS ONE 2021, 16, e0250852. [Google Scholar] [CrossRef]
  9. Rojas-Garbanzo, C.; Rodríguez, L.; Pérez, A.M.; Mayorga-Gross, A.L.; Vásquez-Chaves, V.; Fuentes, E.; Palomo, I. Anti-platelet activity and chemical characterization by UPLC-DAD-ESI-QTOF-MS of the main polyphenols in extracts from Psidium leaves and fruits. Food Res. Int. 2021, 141, 110070. [Google Scholar] [CrossRef]
  10. Alañón, M.E.; Palomo, I.; Rodríguez, L.; Fuentes, E.; Arráez-Román, D.; Segura-Carretero, A. Antiplatelet activity of natural bioactive extracts from mango (Mangifera indica L.) and its by-products. Antioxidants 2019, 8, 517. [Google Scholar] [CrossRef] [Green Version]
  11. Ganesan, K.; Xu, B. Polyphenol-rich dry common beans (Phaseolus vulgaris L.) and their health benefits. Int. J. Mol. Sci. 2017, 18, 2331. [Google Scholar] [CrossRef] [Green Version]
  12. Shang, R.; Wu, H.; Guo, R.; Liu, Q.; Pan, L.; Li, J.; Hu, Z.; Chen, C. The Diversity of Four Anti-nutritional Factors in Common Bean. Hortic. Plant J. 2016, 2, 97–104. [Google Scholar] [CrossRef] [Green Version]
  13. Rodríguez, L.; Mendez, D.; Montecino, H.; Carrasco, B.; Arevalo, B.; Palomo, I.; Fuentes, E. Role of Phaseolus vulgaris L. in the Prevention of Cardiovascular Diseases–Cardioprotective Potential of Bioactive Compounds. Plants 2022, 11, 186. [Google Scholar] [CrossRef] [PubMed]
  14. Alcázar-Valle, M.; Lugo-Cervantes, E.; Mojica, L.; Morales-Hernández, N.; Reyes-Ramírez, H.; Enríquez-Vara, J.N.; García-Morales, S. Bioactive compounds, antioxidant activity, and antinutritional content of legumes: A comparison between four phaseolus species. Molecules 2020, 25, 3528. [Google Scholar] [CrossRef] [PubMed]
  15. Mojica, L.; Meyer, A.; Berhow, M.A.; de Mejía, E.G. Bean cultivars (Phaseolus vulgaris L.) have similar high antioxidant capacity, in vitro inhibition of α-amylase and α-glucosidase while diverse phenolic composition and concentration. Food Res. Int. 2015, 69, 38–48. [Google Scholar] [CrossRef]
  16. Sutivisedsak, N.; Cheng, H.; Willett, J.; Lesch, W.; Tangsrud, R.; Biswas, A. Microwave-assisted extraction of phenolics from bean (Phaseolus vulgaris L.). Food Res. Int. 2010, 43, 516–519. [Google Scholar] [CrossRef]
  17. Al Jitan, S.; Alkhoori, S.A.; Yousef, L.F. Phenolic acids from plants: Extraction and application to human health. Stud. Nat. Prod. Chem. 2018, 58, 389–417. [Google Scholar]
  18. Kumar, M.; Dahuja, A.; Tiwari, S.; Punia, S.; Tak, Y.; Amarowicz, R.; Bhoite, A.G.; Singh, S.; Joshi, S.; Panesar, P.S. Recent trends in extraction of plant bioactives using green technologies: A review. Food Chem. 2021, 353, 129431. [Google Scholar] [CrossRef]
  19. Bubalo, M.C.; Vidović, S.; Redovniković, I.R.; Jokić, S. New perspective in extraction of plant biologically active compounds by green solvents. Food Bioprod. Process. 2018, 109, 52–73. [Google Scholar] [CrossRef]
  20. Azmir, J.; Zaidul, I.S.M.; Rahman, M.M.; Sharif, K.; Mohamed, A.; Sahena, F.; Jahurul, M.; Ghafoor, K.; Norulaini, N.; Omar, A. Techniques for extraction of bioactive compounds from plant materials: A review. J. Food Eng. 2013, 117, 426–436. [Google Scholar] [CrossRef]
  21. Picot-Allain, C.; Mahomoodally, M.F.; Ak, G.; Zengin, G. Conventional versus green extraction techniques—A comparative perspective. Curr. Opin. Food Sci. 2021, 40, 144–156. [Google Scholar] [CrossRef]
  22. Chemat, F.; Vian, M.A.; Cravotto, G. Green extraction of natural products: Concept and principles. Int. J. Mol. Sci. 2012, 13, 8615–8627. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Panja, P. Green extraction methods of food polyphenols from vegetable materials. Curr. Opin. Food Sci. 2018, 23, 173–182. [Google Scholar] [CrossRef]
  24. Pilařová, V.; Al Hamimi, S.; Cunico, L.P.; Nováková, L.; Turner, C. Extending the design space in solvent extraction–from supercritical fluids to pressurized liquids using carbon dioxide, ethanol, ethyl lactate, and water in a wide range of proportions. Green Chem. 2019, 21, 5427–5436. [Google Scholar] [CrossRef] [Green Version]
  25. Osorio-Tobón, J.F. Technology. Recent advances and comparisons of conventional and alternative extraction techniques of phenolic compounds. J. Food Sci. Technol. 2020, 57, 4299–4315. [Google Scholar] [CrossRef]
  26. Destandau, E.; Michel, T.; Elfakir, C. Microwave-assisted extraction. Nat. Nat. Prod. Extr. Princ. Appl. 2013, 21, 113. [Google Scholar]
  27. Zurob, E.; Cabezas, R.; Villarroel, E.; Rosas, N.; Merlet, G.; Quijada-Maldonado, E.; Romero, J.; Plaza, A. Design of natural deep eutectic solvents for the ultrasound-assisted extraction of hydroxytyrosol from olive leaves supported by COSMO-RS. Sep. Purif. Technol. 2020, 248, 117054. [Google Scholar] [CrossRef]
  28. Vinatoru, M.; Mason, T.; Calinescu, I. Ultrasonically assisted extraction (UAE) and microwave assisted extraction (MAE) of functional compounds from plant materials. TrAC Trends Anal. Chem. 2017, 97, 159–178. [Google Scholar] [CrossRef]
  29. Vilkhu, K.; Mawson, R.; Simons, L.; Bates, D. Applications and opportunities for ultrasound assisted extraction in the food industry—A review. Innov. Food Sci. Emerg. Technol. 2008, 9, 161–169. [Google Scholar] [CrossRef]
  30. Yang, Q.-Q.; Gan, R.-Y.; Ge, Y.-Y.; Zhang, D.; Corke, H. Ultrasonic treatment increases extraction rate of common bean (Phaseolus vulgaris L.) antioxidants. Antioxidants 2019, 8, 83. [Google Scholar] [CrossRef] [Green Version]
  31. Rodriguez-Azua, R.; Quinteros, E.F.; Olate-Briones, A.; Moore-Carrasco, R. Phaseolus vulgaris Exerts an Inhibitory Effect on Platelet Aggregation through AKT Dependent Way. Prev. Nutr. Food Sci. 2018, 23, 102–107. [Google Scholar] [CrossRef]
  32. Gupta, R.B.; Shim, J.-J. Solubility in Supercritical Carbon Dioxide; CRC Press: Boca Raton, FL, USA, 2006. [Google Scholar]
  33. Chemat, F.; Rombaut, N.; Sicaire, A.-G.; Meullemiestre, A.; Fabiano-Tixier, A.-S.; Abert-Vian, M. Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrason. Sonochem. 2017, 34, 540–560. [Google Scholar] [CrossRef] [PubMed]
  34. Cassol, L.; Rodrigues, E.; Zapata Noreña, C.P. Extracting phenolic compounds from Hibiscus sabdariffa L. calyx using microwave assisted extraction. Ind. Crops Prod. 2019, 133, 168–177. [Google Scholar] [CrossRef]
  35. Kumar, M.; Dahuja, A.; Sachdev, A.; Kaur, C.; Varghese, E.; Saha, S.; Sairam, K. Evaluation of enzyme and microwave-assisted conditions on extraction of anthocyanins and total phenolics from black soybean (Glycine max L.) seed coat. Int. J. Biol. Macromol. 2019, 135, 1070–1081. [Google Scholar] [CrossRef]
  36. Gil-Martín, E.; Forbes-Hernández, T.; Alejandro, R.; Cianciosi, D.; Giampieri, F.; Battino, M. Influence of the extraction method on the recovery of bioactive phenolic compounds from food industry by-products. Food Chem. 2021, 378, 131918. [Google Scholar] [CrossRef] [PubMed]
  37. Chen, Q.; Dong, W.; Wei, C.; Hu, R.; Long, Y. Combining integrated ultrasonic-microwave technique with ethanol to maximise extraction of green coffee oil from Arabica coffee beans. Ind. Crops Prod. 2020, 151, 112405. [Google Scholar] [CrossRef]
  38. Guo, Z.; Zhao, B.; Li, H.; Miao, S.; Zheng, B. Optimization of ultrasound-microwave synergistic extraction of prebiotic oligosaccharides from sweet potatoes (Ipomoea batatas L.). Innov. Food Sci. Emerg. Technol. 2019, 54, 51–63. [Google Scholar] [CrossRef]
  39. Mellinas, A.C.; Jiménez, A.; Garrigós, M.C. Optimization of microwave-assisted extraction of cocoa bean shell waste and evaluation of its antioxidant, physicochemical and functional properties. LWT 2020, 127, 109361. [Google Scholar] [CrossRef]
  40. Aware, C.B.; Patil, R.R.; Vyavahare, G.D.; Gurme, S.T.; Jadhav, J.P. Ultrasound-assisted aqueous extraction of phenolic, flavonoid compounds and antioxidant activity of Mucuna macrocarpa beans: Response surface methodology optimization. J. Am. Coll. Nutr. 2019, 38, 364–372. [Google Scholar] [CrossRef]
  41. Thaipong, K.; Boonprakob, U.; Crosby, K.; Cisneros-Zevallos, L.; Byrne, D. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J. Food Compos. Anal. 2006, 19, 669–675. [Google Scholar] [CrossRef]
  42. Prior, R.L.; Hoang, H.; Gu, L.; Wu, X.; Bacchiocca, M.; Howard, L.; Hampsch-Woodill, M.; Huang, D.; Ou, B.; Jacob, R.; et al. Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORACFL)) of plasma and other biological and food samples. J. Agric. Food Chem. 2003, 51, 3273–3279. [Google Scholar] [CrossRef]
  43. Xu, B.J.; Chang, S. A comparative study on phenolic profiles and antioxidant activities of legumes as affected by extraction solvents. J. Food Sci. 2007, 72, S159–S166. [Google Scholar] [CrossRef] [PubMed]
  44. Zhou, Y.; Xu, X.-Y.; Gan, R.-Y.; Zheng, J.; Li, Y.; Zhang, J.-J.; Xu, D.-P.; Li, H.-B. Optimization of ultrasound-assisted extraction of antioxidant polyphenols from the seed coats of red sword bean (Canavalia gladiate (Jacq.) DC.). Antioxidants 2019, 8, 200. [Google Scholar] [CrossRef] [Green Version]
  45. Rodríguez Madrera, R.; Campa Negrillo, A.; Suárez Valles, B.; Ferreira Fernández, J. Phenolic Content and Antioxidant Activity in Seeds of Common Bean (Phaseolus vulgaris L.). Foods 2021, 10, 864. [Google Scholar] [CrossRef] [PubMed]
  46. Aparicio-Fernandez, X.; Yousef, G.G.; Loarca-Pina, G.; De Mejia, E.; Lila, M.A. Characterization of polyphenolics in the seed coat of Black Jamapa bean (Phaseolus vulgaris L.). J. Agric. Food Chem. 2005, 53, 4615–4622. [Google Scholar] [CrossRef]
  47. Nakashima, S.; Koike, T.; Nozawa, Y. Genistein, a protein tyrosine kinase inhibitor, inhibits thromboxane A2-mediated human platelet responses. Mol. Pharmacol. 1991, 39, 475–480. [Google Scholar]
  48. Luceri, C.; Giannini, L.; Lodovici, M.; Antonucci, E.; Abbate, R.; Masini, E.; Dolara, P. p-Coumaric acid, a common dietary phenol, inhibits platelet activity in vitro and in vivo. Br. J. Nutr. 2007, 97, 458–463. [Google Scholar] [CrossRef] [Green Version]
  49. Ostertag, L.M.; O’Kennedy, N.; Horgan, G.W.; Kroon, P.A.; Duthie, G.G.; de Roos, B. In vitro anti-platelet effects of simple plant-derived phenolic compounds are only found at high, non-physiological concentrations. Mol. Nutr. Food Res. 2011, 55, 1624–1636. [Google Scholar] [CrossRef] [PubMed]
  50. Fuentes, E.; Caballero, J.; Alarcón, M.; Rojas, A.; Palomo, I. Chlorogenic acid inhibits human platelet activation and thrombus formation. PLoS ONE 2014, 9, e90699. [Google Scholar] [CrossRef] [Green Version]
  51. Cho, H.-J.; Kang, H.-J.; Kim, Y.-J.; Lee, D.-H.; Kwon, H.-W.; Kim, Y.-Y.; Park, H.-J. Inhibition of platelet aggregation by chlorogenic acid via cAMP and cGMP-dependent manner. Blood Coagul. Fibrinolysis 2012, 23, 629–635. [Google Scholar] [CrossRef]
  52. Hong, Q.; Ma, Z.-C.; Huang, H.; Wang, Y.-G.; Tan, H.-L.; Xiao, C.-R.; Liang, Q.-D.; Zhang, H.-T.; Gao, Y. Antithrombotic activities of ferulic acid via intracellular cyclic nucleotide signaling. Eur. J. Pharmacol. 2016, 777, 1–8. [Google Scholar] [CrossRef]
  53. Prince, P.S.M.; Roy, A.J. p-Coumaric acid attenuates apoptosis in isoproterenol-induced myocardial infarcted rats by inhibiting oxidative stress. Int. J. Cardiol. 2013, 168, 3259–3266. [Google Scholar] [CrossRef] [PubMed]
  54. Abdel-Wahab, M.; El-Mahdy, M.; Abd-Ellah, M.; Helal, G.; Khalifa, F.; Hamada, F. Influence of p-coumaric acid on doxorubicin-induced oxidative stress in rat’s heart. Pharmacol. Res. 2003, 48, 461–465. [Google Scholar] [CrossRef]
  55. Wan, Y.J.; Zhuang, P.W.; Zhang, Y.J. Antithrombotic activity of Formononetin sodium and its mechanism. Chin. J. New Drugs 2016, 25, 1355–1358. [Google Scholar]
  56. Landolfi, R.; Mower, R.L.; Steiner, M. Modification of platelet function and arachidonic acid metabolism by bioflavonoids: Structure-activity relations. Biochem. Pharmacol. 1984, 33, 1525–1530. [Google Scholar] [CrossRef]
  57. Zang, B.; Jin, M.; Wu, W.; Chen, W.; Piao, Y.; Li, J. Antagonistic effect of myricetin on platelet activing factor. Cancer Metastasis Rev. 2003, 38, 831–833. [Google Scholar]
  58. Gottstein, N.; Ewins, B.A.; Eccleston, C.; Hubbard, G.P.; Kavanagh, I.C.; Minihane, A.-M.; Weinberg, P.D.; Rimbach, G. Effect of genistein and daidzein on platelet aggregation and monocyte and endothelial function. Br. J. Nutr. 2003, 89, 607–615. [Google Scholar] [CrossRef]
  59. Zhu, H.; Zou, L.; Tian, J.; Lin, F.; He, J.; Hou, J. Protective effects of sulphonated formononetin in a rat model of cerebral ischemia and reperfusion injury. Plant Med. 2014, 80, 262–268. [Google Scholar] [CrossRef] [Green Version]
  60. Ozaki, Y.; Yatomi, Y.; Jinnai, Y.; Kume, S. Effects of genistein, a tyrosine kinase inhibitor, on platelet functions: Genistein attenuates thrombin-induced Ca2+ mobilization in human platelets by affecting polyphosphoinositide turnover. Biochem. Pharmacol. 1993, 46, 395–403. [Google Scholar] [CrossRef]
  61. Dhar, A.; Paul, A.; Shukla, S. Platelet-activating factor stimulation of tyrosine kinase and its relationship to phospholipase C in rabbit platelets: Studies with genistein and monoclonal antibody to phosphotyrosine. Mol. Pharmacol. 1990, 37, 519–525. [Google Scholar]
  62. Chtourou, Y.; Kamoun, Z.; Zarrouk, W.; Kebieche, M.; Kallel, C.; Gdoura, R.; Fetoui, H. Naringenin ameliorates renal and platelet purinergic signalling alterations in high-cholesterol fed rats through the suppression of ROS and NF-κB signaling pathways. Food Funct. 2016, 7, 183–193. [Google Scholar] [CrossRef]
  63. Yang, Y.; Shi, Z.; Reheman, A.; Jin, J.W.; Li, C.; Wang, Y.; Andrews, M.C.; Chen, P.; Zhu, G.; Ling, W. Plant food delphinidin-3-glucoside significantly inhibits platelet activation and thrombosis: Novel protective roles against cardiovascular diseases. PLoS ONE 2012, 7, e37323. [Google Scholar] [CrossRef] [Green Version]
  64. Yao, Y.; Chen, Y.; Adili, R.; McKeown, T.; Chen, P.; Zhu, G.; Li, D.; Ling, W.; Ni, H.; Yang, Y. Plant-based food cyanidin-3-glucoside modulates human platelet glycoprotein VI signaling and inhibits platelet activation and thrombus formation. J. Nutr. 2017, 147, 1917–1925. [Google Scholar] [CrossRef] [Green Version]
  65. Song, F.; Zhu, Y.; Shi, Z.; Tian, J.; Deng, X.; Ren, J.; Andrews, M.C.; Ni, H.; Ling, W.; Yang, Y. Plant food anthocyanins inhibit platelet granule secretion in hypercholesterolaemia: Involving the signalling pathway of PI3K–Akt. Thromb. Haemost. 2014, 112, 981–991. [Google Scholar] [CrossRef]
  66. Zang, L.-Y.; Cosma, G.; Gardner, H.; Shi, X.; Castranova, V.; Vallyathan, V. Effect of antioxidant protection by p-coumaric acid on low-density lipoprotein cholesterol oxidation. Am. J. Physiol.-Cell Physiol. 2000, 279, C954–C960. [Google Scholar] [CrossRef] [Green Version]
  67. Aboonabi, A.; Meyer, R.R.; Gaiz, A.; Singh, I. Anthocyanins in berries exhibited anti-atherogenicity and antiplatelet activities in a metabolic syndrome population. Nutr. Res. 2020, 76, 82–93. [Google Scholar] [CrossRef]
  68. Fuentes, E.; Wehinger, S.; Trostchansky, A. Regulation of Key Antiplatelet Pathways by Bioactive Compounds with Minimal Bleeding Risk. Int. J. Mol. Sci. 2021, 22, 12380. [Google Scholar] [CrossRef]
  69. Méndez, D.; Arauna, D.; Fuentes, F.; Araya-Maturana, R.; Palomo, I.; Alarcón, M.; Sebastián, D.; Zorzano, A.; Fuentes, E. Mitoquinone (MitoQ) inhibits platelet activation steps by reducing ROS levels. Int. J. Mol. Sci. 2020, 21, 6192. [Google Scholar] [CrossRef]
  70. Fuentes, E.; Palomo, I. Relationship between platelet PPARs, cAMP Levels, and P-selectin expression: Antiplatelet activity of natural products. Evid.-Based Complement. Altern. Med. 2013, 2013, 861786. [Google Scholar] [CrossRef] [Green Version]
  71. Fuentes, E.; Palomo, I. Antiplatelet effects of natural bioactive compounds by multiple targets: Food and drug interactions. J. Funct. Foods 2014, 6, 73–81. [Google Scholar] [CrossRef]
  72. González, B.; Vogel, H.; Razmilic, I.; Wolfram, E. Polyphenol, anthocyanin and antioxidant content in different parts of maqui fruits (Aristotelia chilensis) during ripening and conservation treatments after harvest. Ind. Crops Prod. 2015, 76, 158–165. [Google Scholar] [CrossRef]
  73. Céspedes, C.L.; El-Hafidi, M.; Pavon, N.; Alarcon, J. Antioxidant and cardioprotective activities of phenolic extracts from fruits of Chilean blackberry Aristotelia chilensis (Elaeocarpaceae), Maqui. Food Chem. 2008, 107, 820–829. [Google Scholar] [CrossRef]
  74. Martino, H.S.D.; Bigonha, S.M.; Cardoso, L.D.M.; Rosa, C.D.O.B.; Costa, N.M.B.; Cárdenas, L.D.L.Á.R.; Ribeiro, S.M.R. Nutritional and bioactive compounds of bean: Benefits to human health. In Hispanic Foods: Chemistry and Bioactive Compounds; ACS Publications: Washington, DC, USA, 2012; pp. 233–258. [Google Scholar]
  75. Llompart, M.; Celeiro, M.; Dagnac, T. Microwave-assisted extraction of pharmaceuticals, personal care products and industrial contaminants in the environment. TrAC Trends Anal. Chem. 2019, 116, 136–150. [Google Scholar] [CrossRef]
  76. Ahmad-Qasem, M.H.; Cánovas, J.; Barrajón-Catalán, E.; Micol, V.; Cárcel, J.A.; García-Pérez, J.V. Kinetic and compositional study of phenolic extraction from olive leaves (var. Serrana) by using power ultrasound. Innov. Food Sci. Emerg. Technol. 2013, 17, 120–129. [Google Scholar] [CrossRef]
  77. Lee, Y.S.; Lee, S.; Lee, H.S.; Kim, B.-K.; Ohuchi, K.; Shin, K.H. Inhibitory effects of isorhamnetin-3-O-β-D-glucoside from Salicornia herbacea on rat lens aldose reductase and sorbitol accumulation in streptozotocin-induced diabetic rat tissues. Biol. Pharm. Bull. 2005, 28, 916–918. [Google Scholar] [CrossRef] [Green Version]
  78. Zapata Díez, C.D.; Zapata Ocampo, P.A. Estandarización del Método ORAC Como Herramienta Básica de Análisis de la Capacidad Antioxidante de Diversas Sustancias. 2019. Available online: repository.ces.edu.co (accessed on 5 April 2022).
  79. Ou, B.; Chang, T.; Huang, D.; Prior, R.L. Determination of total antioxidant capacity by oxygen radical absorbance capacity (ORAC) using fluorescein as the fluorescence probe: First action 2012.23. J. AOAC Int. 2013, 96, 1372–1376. [Google Scholar] [CrossRef]
  80. Rodriguez, L.; Badimon, L.; Mendez, D.; Padro, T.; Vilahur, G.; Pena, E.; Carrasco, B.; Vogel, H.; Palomo, I.; Fuentes, E. Antiplatelet Activity of Isorhamnetin via Mitochondrial Regulation. Antioxidants 2021, 10, 666. [Google Scholar] [CrossRef]
  81. Mendez, D.; Urra, F.A.; Millas-Vargas, J.P.; Alarcon, M.; Rodriguez-Lavado, J.; Palomo, I.; Trostchansky, A.; Araya-Maturana, R.; Fuentes, E. Synthesis of antiplatelet ortho-carbonyl hydroquinones with differential action on platelet aggregation stimulated by collagen or TRAP-6. Eur. J. Med. Chem. 2020, 192, 112187. [Google Scholar] [CrossRef]
  82. Mendez, D.; Donoso-Bustamante, V.; Pablo Millas-Vargas, J.; Pessoa-Mahana, H.; Araya-Maturana, R.; Fuentes, E. Synthesis and pharmacological evaluation of acylhydroquinone derivatives as potent antiplatelet agents. Biochem. Pharmacol. 2021, 183, 114341. [Google Scholar] [CrossRef]
  83. Rojas-Garbanzo, C. Psidium Fruits: Endemic fruits of latin America with a wide variety of phytochemicals. Ann. Nutr. Food Sci. 2018, 2, 1016. [Google Scholar]
  84. Yang, Q.Q.; Gan, R.Y.; Ge, Y.Y.; Zhang, D.; Corke, H. Polyphenols in common beans (Phaseolus vulgaris L.): Chemistry, analysis, and factors affecting composition. Compr. Rev. Food Sci. Food Saf. 2018, 17, 1518–1539. [Google Scholar] [CrossRef] [Green Version]
  85. Ombra, M.N.; d’Acierno, A.; Nazzaro, F.; Riccardi, R.; Spigno, P.; Zaccardelli, M.; Pane, C.; Maione, M.; Fratianni, F. Phenolic composition and antioxidant and antiproliferative activities of the extracts of twelve common bean (Phaseolus vulgaris L.) endemic ecotypes of Southern Italy before and after cooking. Oxidative Med. Cell. Longev. 2016, 2016, 1398298. [Google Scholar] [CrossRef] [Green Version]
  86. Singh, B.; Singh, N.; Thakur, S.; Kaur, A. Ultrasound assisted extraction of polyphenols and their distribution in whole mung bean, hull and cotyledon. J. Food Sci. Technol. 2017, 54, 921–932. [Google Scholar] [CrossRef] [Green Version]
  87. Luo, J.; Cai, W.; Wu, T.; Xu, B. Phytochemical distribution in hull and cotyledon of adzuki bean (Vigna angularis L.) and mung bean (Vigna radiate L.), and their contribution to antioxidant, anti-inflammatory and anti-diabetic activities. Food Chem. 2016, 201, 350–360. [Google Scholar] [CrossRef]
Plants 11 01179 g001 550
Figure 1. Cytotoxicity on platelets. The results were obtained by LDH release from supernatant on washed platelets and measured at 490 nm in a microplate reader. Data expressed as mean ± SD, n = 6 independent trials; Vehicle = PBS. Triton X-100 as a positive control (maximum LDH release). The statistical analysis was performed using the ANOVA (Dunnet test). *** p < 0.001 vs. vehicle.
Figure 1. Cytotoxicity on platelets. The results were obtained by LDH release from supernatant on washed platelets and measured at 490 nm in a microplate reader. Data expressed as mean ± SD, n = 6 independent trials; Vehicle = PBS. Triton X-100 as a positive control (maximum LDH release). The statistical analysis was performed using the ANOVA (Dunnet test). *** p < 0.001 vs. vehicle.
Plants 11 01179 g001
Plants 11 01179 g002 550
Figure 2. Platelet inhibition of more active extracts of P. vulgaris L. The more active extracts, with the percentage of platelet inhibition above 50%, were selected to evaluate the concentration necessary to reduce platelet aggregation by 50% (IC50), determined from the concentration curves of the extracts of common bean at 0.025, 0.05, 0.10, 0.25, 0.50, 0.75 and 1.00 mg/mL.
Figure 2. Platelet inhibition of more active extracts of P. vulgaris L. The more active extracts, with the percentage of platelet inhibition above 50%, were selected to evaluate the concentration necessary to reduce platelet aggregation by 50% (IC50), determined from the concentration curves of the extracts of common bean at 0.025, 0.05, 0.10, 0.25, 0.50, 0.75 and 1.00 mg/mL.
Plants 11 01179 g002
Table
Table 1. Results of the content of soluble solids, total phenols and antioxidant activity of 25 landraces of P. vulgaris L.
Table 1. Results of the content of soluble solids, total phenols and antioxidant activity of 25 landraces of P. vulgaris L.
LandracesExtraction TechniqueSoluble Solid ContentTPC (mg GAE/g)ORAC μmolTE/gTRAP-6 10 µM
PA (%)		ADP 4 µM
PA (%)
AraucoMAE1.50 ± 0.00 a572.58 ± 10.21 a22,725.36 ± 691.51 a26.8 ± 1.2 a71.3 ± 1.5 a
UAE1.33 ± 0.06 a464.94 ± 10.36 a9048.36 ± 289.97 b49.1 ± 3.5 b71.0 ± 2.5 a
Blanco EspañolMAE0.93 ± 0.06 b241.94 ± 18.17 b4709.13 ± 1233.93 c23.8 ± 2.6 a75.7 ± 2.2 ab
UAE0.97 ± 0.06 b280.27 ± 46.02 b2721.33 ± 257.49 c36.2 ± 3.0 ab71.5 ± 3.0 a
BomberoMAE1.00 ± 0.17 b311.73 ± 4.42 ba3775.75 ± 966.34 c77.5 ± 3.1 c92.3 ± 0.8 cb
UAE0.90 ± 0.06 b227 ± 22.95 b2821.35 ± 70.83 c17.8 ± 2.4 a71.5 ± 2.4 a
CabritaMAE1.00 ± 0.10 b264.66 ± 11.62 ba8045.22 ± 600.05 b36.3 ± 3.1 ab84.7 ± 0.8 b
UAE1.27 ± 0.06 ab437.89 ± 36.74 a14,489.49 ± 344.15 db65.0 ± 3.2 cb87.9 ± 1.4 b
CimarrónMAE2.03 ± 0.06 c520.47 ± 31.28 a2722.64 ± 932.78 c79.1 ± 3.4 c73.8 ± 3.8 a
UAE2.27 ± 0.06 c352.27 ± 34.03 ba9612.22 ± 534.08 b78.6 ± 2.4 c84.2 ± 1.3 b
CisneMAE1.07 ± 0.06 ab220.08 ± 11.98 b3657.96 ± 386.98 c60.1 ± 4.1 cb80.8 ± 2.5 ba
UAE0.87 ± 0.06 b246.85 ± 17.66 ba3310.31 ± 301.25 c72.4 ± 3.7 c86.5 ± 1.2 b
CoscorrónMAE1.27 ± 0.06 ab280.06 ± 38.53 ba2704.07 ± 482.92 c7.9 ± 0.4 d73.8 ± 1.9 a
UAE0.97 ± 0.06 b225.19 ± 11.30 baNd54.9 ± 1.0 b84.8 ± 0.8 b
FrutillaMAE1.10 ± 0.00 ba414.45 ± 29.54 a21,630.05 ± 260.90 a21.3 ± 1.1 a82.1 ± 1.8 b
UAE1.07 ± 0.06 b383.32 ± 39.47 ba14,238.93 ± 910.50 db50.3 ± 1.7 b85.4 ± 0.6 b
GansoMAE1.70 ± 0.10 a431.83 ± 34.86 a9375.30 ± 1722.49 b17.1 ± 2.7 a74.1 ± 1.2 a
UAE1.47 ± 0.06 a318.96 ± 27.29 ba3313.60 ± 351.80 c69.8 ± 3.9 c82.3 ± 0.7 b
Hallado AlemánMAE1.00 ± 0.00 b541.85 ± 33.37 a17,682.97 ± 248.60 d85.8 ± 0.7 c81.8 ± 1.2 b
UAE0.93 ± 0.15 b337.91 ± 19.47 ba11,321.46 ± 987.80 bd11.0 ± 0.9 ad73.3 ± 2.9 a
LunatusMAE1.53 ± 0.06 a283.09 ± 69.11 ba5857.60 ± 2350.40 b86.8 ± 0.8 c90.8 ± 0.4 cb
UAE1.47 ± 0.12 a298.27 ± 26.91 ba797.13 ± 158.30 c7.8 ± 1.4 d72.6 ± 1.6 a
MantecaMAE1.53 ± 0.06 a603.89 ± 6.83 ac12,561.54 ± 1096.83 bd55.2 ± 1.3 b79.8 ± 1.4 ab
UAE1.00 ± 0.00 b321.96 ± 5.92 ba2867.47 ± 214.95 c78.5 ± 2.9 c87.9 ± 1.2 bc
MantequillaMAE1.50 ± 0.00 a308.89 ± 9.40 ba13,721.58 ± 3113.94 db22.3 ± 1.8 a76.7 ± 2.4 ab
UAE0.93 ± 0.06 b340.85 ± 22.87 ba3976.00 ± 463.28 c56.3 ± 1.3 b76.4 ± 1.8 ab
NegroMAE1.30 ± 0.00 a806.90 ± 6.66 c16,659.61 ± 158.45 d7.8 ± 1.2 d73.6 ± 2.0 a
UAE1.00 ± 0.10 b543.40 ± 12.02 a7243.07 ± 964.23 b25.3 ± 1.4 a84.0 ± 0.9 bc
Negro AraucoMAE1.27 ± 0.02 a345.89 ± 47.58 ba8955.42 ± 516.75 b28.5 ± 2.8 a72.8 ± 2.0 a
UAE1.17 ± 0.02 ba323.62 ± 77.94 ba3481.76 ± 759.27 c47.0 ± 1.7 b78.0 ± 1.4 ab
PajaritoMAE1.33 ± 0.06 a594.36 ± 8.80 a18,723.60 ± 1700.55 ad56.7 ± 0.9 b84.1 ± 1.1 bc
UAE1.23 ± 0.06 a485.70 ± 18.84 a12,904.29 ± 968.70 bd78.0 ± 2.8 c87.7 ± 1.3 bc
PaloMAE1.30 ± 0.10 a423.06 ± 27.19 a17,828.49 ± 826.24 d85.4 ± 0.3 c81.0 ± 3.3 bc
UAE1.33 ± 0.06 a391.23 ± 50.86 ba16,179.95 ± 2142.43 d23.5 ± 1.5 a71.1 ± 3.4 a
Pallar ManchadoMAE0.93 ± 0.06 b316.52 ± 93.50 ba15,983.88 ± 674.45 d20.9 ± 1.0 a88.0 ± 1.3 bc
UAE0.90 ± 0.00 b374.63 ± 53.69 ba9331.82 ± 891.31 b18.7 ± 1.1 a74.0 ± 0.6 a
Pallar MoradoMAE1.17 ± 0.06 ab795.95 ± 29.97 c11,233.61 ± 1702.32 b31.9 ± 0.6 ab73.5 ± 2.6 a
UAE1.03 ± 0.06 b495.63 ± 39.09 a10,993.16 ± 476.40 b20.8 ± 1.0 a68.0 ± 1.6 a
PeumoMAE1.40 ± 0.10 a866.17 ± 30.28 c25,642.98 ± 2396.79 a46.1 ± 6.2 b81.8 ± 1.9 b
UAE0.90 ± 0.00 b720.52 ± 18.43 c22,596.47 ± 3452.67 a81.7 ± 1.7 c82.9 ± 0.5 b
RojoMAE1.00 ± 0.00 b590.86 ± 24.25 a12,809.11 ± 976.94 b35.5 ± 2.7 ab75.0 ± 2.0 ba
UAE0.90 ± 0.10 b518.22 ± 11.90 a6535.65 ± 1872.82 b57.1 ± 7.5 bc69.0 ± 7.0 a
SapitoMAE1.17 ± 0.06 ab265.30 ± 12.93 ba5184.58 ± 359.74 b69.6 ± 4.7 c69.0 ± 4.8 a
UAE1.13 ± 0.06 ab232.68 ± 4.14 ba4345.33 ± 26.73 c79.4 ± 3.0 c87.8 ± 0.9 bc
SojaMAE1.83 ± 0.06 a893.45 ± 87.30 c35,642.85 ± 2588.88 e22.4 ± 2.8 a75.2 ± 2.2 ab
UAE1.63 ± 0.06 a696.85 ± 21.06 ca19,820.36 ± 1474.80 a28.1 ± 2.9 a74.0 ± 1.9 a
TorcazaMAE1.57 ± 0.06 a349.19 ± 16.56 ab5720.66 ± 145.28 b27.5 ± 2.0 a78.7 ± 3.4 ab
UAE1.63 ± 0.12 a302.92 ± 15.42 ba3337.06 ± 888.64 c48.0 ± 0.7 b66.2 ± 3.9 a
TórtolaMAE1.90 ± 0.00 c327.65 ± 24.89 ba13,617.59 ± 1519.75 bd12.0 ± 1.3 da78.2 ± 1.7 ab
UAE1.00 ± 0.10 b284.80 ± 6.42 ba3561.15 ± 880.60 c75.06 ± 2.9 c80.8 ± 1.2 b
Data are expressed as mean ± SD, n = 3 from at least three independent experiments. Nd: unidentified (value was below the limit of quantification of the calibration curve). Different superscript letters (a–e) in the same column show significant differences within each landrace, according to Tukey’s test (p < 0.05).
Table
Table 2. Total monomeric anthocyanin on extracts.
Table 2. Total monomeric anthocyanin on extracts.
ExtractsCyanidin-3-glucoside (mg/100 g of Sample)
MAE Pallar Morado25.81 ± 0.02
MAE Ganso24.86 ± 0.04
UAE Bombero23.58 ± 0.03
MAE Tortola12.72 ± 0.02
MAE Soja12.33 ± 0.03
UAE Palo10.58 ± 0.01
MAE Pallar Manchado9.83 ± 0.01
MAE Negro Arauco9.27 ± 0.01
UAE negro4.42 ± 0.01
UAE Blanco Español0.86 ± 0.00
Table
Table 3. Compounds identified on P. vulgaris L. with cardioprotective potential.
Table 3. Compounds identified on P. vulgaris L. with cardioprotective potential.
Bioactive CompoundCardioprotective Effect and/or MechanismsReference
Phenolic acids
(coumaric, chlorogenic, ferulic, cinnamic, sinapic, protocatechuic, vanillic, syringic acid)		Inhibit platelet adhesion and/or aggregation[48,49,50,51,52]
Inhibit LDL oxidation[66]
Protects the heart against oxidative stress[53,54]
Inhibits platelet inflammatory mediators[50]
Increases cAMP levels[50,51,52]
Inhibit P-selectin expression[49,53]
activation of cAMP and cGMP signaling pathways[50,51,52]
Flavonoids
(Myricerin, Genistein, Glycitein, Formononetin, Naringenin, Hesperetin, Daidzein, Catechin, Kaemferol-3-glucoside, Taxifolin, Apigenin-7 Glycoside, Luteolin-7 Glycoside)		Inhibit platelet aggregation induced by several agonists[55,56,57,58]
Inhibit PI3 production[59,60,61]
Protect against cerebral ischemia and reperfusion injury in rats and enhances cerebrovascular angiogenesis in human umbilical vein endothelial cells[59]
Inhibits levels of inflammatory markers[62]
Increases cAMP levels[56]
Anthocyanins
(Delphinidin, Petunidin, Malvidin, Cyanidin-3-O-glucoside, Pelargonidin-3-O-glucoside)		Inhibit platelet aggregation induced by several agonists[63,67]
Inhibit the secretion of alpha and dense granules[65]
Inhibit PI3K/Akt activation, eNOS phosphorylation and cGMP production[63]
Inhibit P-selectin expression[63,64,65]
Reprinted with permission from ref. [13]. Rodríguez, L., et al., 2022.
Table
Table 4. Description of the landraces of P. vulgaris L.
Table 4. Description of the landraces of P. vulgaris L.
Chilean Phaseolus vulgaris L. LandracesCountryRegionVillageGPS Coordinate
Bombero, Cimarrón, Cisne, Ganso, Tórtola, LunatusChileMauleCurepto, Maule−34°.99′09″34,
−72°.01′56″31
Arauco, Palo, Pallar Manchado, Pallar Morado, Negro Arauco, Hallado Alemán, MantecaChileMauleHualañe−35°.01′44″19,
−71°.73′94″57
Cabrita, Coscorrón, Pajarito, Frutilla, Peumo, Rojo, SapitoChileÑubleChanco, Cobquecura−36°.28′66″29,
−72°.71′92″36
Blanco Español, Torcaza, Negro, MantequillaChileMauleLongaví−35°.96′80″49,
−71°.71′36″48
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Rodríguez, L.; Plaza, A.; Méndez, D.; Carrasco, B.; Tellería, F.; Palomo, I.; Fuentes, E. Antioxidant Capacity and Antiplatelet Activity of Aqueous Extracts of Common Bean (Phaseolus vulgaris L.) Obtained with Microwave and Ultrasound Assisted Extraction. Plants 2022, 11, 1179. https://doi.org/10.3390/plants11091179

AMA Style

Rodríguez L, Plaza A, Méndez D, Carrasco B, Tellería F, Palomo I, Fuentes E. Antioxidant Capacity and Antiplatelet Activity of Aqueous Extracts of Common Bean (Phaseolus vulgaris L.) Obtained with Microwave and Ultrasound Assisted Extraction. Plants. 2022; 11(9):1179. https://doi.org/10.3390/plants11091179

Chicago/Turabian Style

Rodríguez, Lyanne, Andrea Plaza, Diego Méndez, Basilio Carrasco, Francisca Tellería, Iván Palomo, and Eduardo Fuentes. 2022. "Antioxidant Capacity and Antiplatelet Activity of Aqueous Extracts of Common Bean (Phaseolus vulgaris L.) Obtained with Microwave and Ultrasound Assisted Extraction" Plants 11, no. 9: 1179. https://doi.org/10.3390/plants11091179

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop