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Phenolic composition and antioxidant potential of grain legume seeds: A review

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DOI: 10.1016/j.foodres.2017.09.026

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Accepted Manuscript

Phenolic composition and antioxidant potential of grain legume


seeds: A review

Balwinder Singh, Jatinder Pal Singh, Amritpal Kaur, Narpinder


Singh

PII: S0963-9969(17)30593-8
DOI: doi: 10.1016/j.foodres.2017.09.026
Reference: FRIN 6975
To appear in: Food Research International
Received date: 20 June 2017
Revised date: 4 September 2017
Accepted date: 8 September 2017

Please cite this article as: Balwinder Singh, Jatinder Pal Singh, Amritpal Kaur, Narpinder
Singh , Phenolic composition and antioxidant potential of grain legume seeds: A review,
Food Research International (2017), doi: 10.1016/j.foodres.2017.09.026

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ACCEPTED MANUSCRIPT
Phenolic composition and antioxidant potential of grain

legume seeds: A review


Balwinder Singh1, Jatinder Pal Singh2, Amritpal Kaur2*, Narpinder Singh2
1
Department of Biotechnology, Khalsa College, Amritsar-143002, Punjab, India
2
Department of Food Science and Technology, Guru Nanak Dev University, Amritsar -143005 Punjab, India

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Email: amritft33@yahoo.co.in

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Abstract

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Legumes are a good source of bioactive phenolic compounds which play significant roles in

many physiological as well as metabolic processes. Phenolic acids, flavonoids and condensed tannins

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are the primary phenolic compounds that are present in legume seeds. Majority of the phenolic
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compounds are present in the legume seed coats. The seed coat of legume seeds primarily contains

phenolic acids and flavonoids (mainly catechins and procyanidins). Gallic and protocatechuic acids
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are common in kidney bean and mung bean. Catechins and procyanidins represent almost 70% of
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total phenolic compounds in lentils and cranberry beans (seed coat). The antioxidant activity of

phenolic compounds is in direct relation with their chemical structures such as number as well as
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position of the hydroxyl groups. Processing mostly leads to the reduction of phenolic compounds in
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legumes owing to chemical rearrangements. Phenolic content also decreases due to leaching of water-

soluble phenolic compounds into the cooking water. The health benefits of phenolic compounds
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include acting as anticarcinogenic, anti-thrombotic, anti-ulcer, anti-artherogenic, anti-allergenic, anti-

inflammatory, antioxidant, immunemodulating, anti-microbial, cardioprotective and analgesic agents.

This review provides comprehensive information of phenolic compounds identified in grain legume

seeds along with discussing their antioxidant and health promoting activities.

Keywords

Legume seeds; Phenolic compounds; Antioxidant activity; Processing; Health benefits

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1. Introduction

Legume seeds constitute an essential part of the human diet as they are excellent sources of

proteins, minerals, vitamins and bioactive compounds (Magalhães et al., 2017). There has been a

significant increase in their production worldwide and the food processing industry is developing

newer uses of these seeds for producing food products having beneficial effect on human health

(Aguilera et al., 2011a). The worldwide production, area harvested and yield of different legume

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seeds is presented in Supplementary Figure S1. Legumes are a good source of bioactive phenolic

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compounds for humans as they play a significant role in many physiological and metabolic processes.

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Additionally, the diets in developing countries are primarily based on legume and cereal products and

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recently there has been an increasing interest in following strictly vegetarian diets among Western

societies (Sabaté and Soret, 2014). Most of the phenolic compounds are concentrated in the seed
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coats of the legumes (Amarowicz and Shahidi, 2017; de Mejıa et al. 1999; Dueñas et al., 2006; Gan et

al., 2016). These compounds function as bioactive compounds and are important determinants of
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color, taste and flavor of foods. They exhibit free radical-scavenging capacity and the ability to
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interact with proteins. The bioactive phenolic compounds present in grain legumes (as reactive

metabolites and associated antioxidant activity) make them suitable candidates for creating new
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functional foods (Aguilera et al., 2011). Phenolic compounds are polyhydroxylated compounds,
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constituting one of the most extensive groups of chemicals present in plant kingdom. These show

structural diversity ranging from simple phenolics to complex as well as highly polymerized
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compounds. The high-molecular weight phenolic compounds having a complex structure are often

referred to as polyphenols. These exhibit plenty of biologically significant functions such as

protection against oxidative stresses and degenerative diseases. These compounds might offer indirect

protection by the activation of endogenous defense systems and with the modulation of cellular

signaling processes. Bioactivities (the specific effect produced in human body upon exposure to

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bioactive compounds) of phenolic compounds exemplify their importance in food products. They

have many health benefits, some of which are acting as anticarcinogenic, anti-thrombotic, anti-ulcer,

anti-artherogenic, anti-allergenic, anti-inflammatory, antioxidant, immunemodulating, anti-microbial,

cardioprotective, vasodialatory and analgesic agents (Balasundram et al., 2006). Alshikh et al. (2015)

documented that phenolic compounds present in legumes not only contributed to their organoleptic

properties, but also provided several health benefits. While surveying the literature, collective

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information in the form of a review on the phenolic compounds (including their antioxidant activities

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as well as health benefits) present in legume seeds is scarce. This review provides comprehensive

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collective information of phenolic compounds reported in different grain legume seeds such as

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common beans (Phaseolus vulgaris), faba bean (Vicia faba), mung bean (Vigna radiate), Pinto bean

(Phaseolus vulgaris), kidney bean (Phaseolus vulgaris L.), adzuki bean (Vigna angularis), lentil
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(Lens culinaris), chickpea (Cicer arietinum), cowpea (Vigna unguiculata), pigeon pea (Cajanus

cajan), pea (Pisum sativum), soybean (Glycine max) and peanut (Arachis hypogaea). Moreover,
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antioxidant and health promoting activities such as reducing the incidence of cancer, heart problems,
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having neuroprotective effects, anti-inflammatory activity, management of diabetes and obesity of

these compounds present in legumes are also discussed.


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2. Phenolic compounds
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Phenolic compounds are widely distributed bioactive secondary metabolites present in all

higher plants that are mainly synthesized by the shikimic acid, pentose phosphate and
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phenylpropanoid pathways (Balasundram et al., 2006). Structurally, they have one or more hydroxyl

groups attached directly to the aromatic ring and can vary from simple molecules to highly complex

polymers (Haminiuk et al., 2012; Singh et al., 2015). Phenolic compounds are divided into subgroups

of phenolic acids, flavonoids, tannins and stilbenes on the basis of number of phenolic hydroxyl

groups attached and structural elements that link benzene rings (Singh et al., 2016b). It is estimated

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that more than 8000 phenolic compounds have been isolated and described in plants (Ouchemoukh et

al., 2016). Phenolic compounds affect the sensory properties of foods and tannins primarily

contribute to the astringency of foods (Landete et al., 2012; Singh et al., 2016a). The flavonoids

include flavonols, flavones, flavanols, flavanones, anthocyanidins and isoflavones. Tannins occur in

complexes with polysaccharides, proteins and alkaloids and are subdivided into hydrolysable and

condensed tannins. Some of these compounds are water soluble (phenolic acids and flavonoids),

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while some are insoluble (some condensed tannins). Flavonoids (60%) and phenolic acids (30%)

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mainly account for phenolic compounds in our diet (Haminiuk et al., 2012).

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There are many available methods for the extraction of phenolic compounds but no single

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method can be categorized as a standard method. The extraction of phenolic compounds is mainly

affected by several parameters (such as the particle size of the samples, the type of solvent used, the
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solute to solvent ratio, agitation rate, mass transfer efficiency and temperature used). The most

important step is to select the optimum conditions required for extraction. Besides this, the crucial
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step in the quantification of these compounds is the preparation method of samples as it influences
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both the accuracy as well as the repeatability during the analysis (Zhao et al., 2011). Finely powdered

samples considerably improve the extraction of phenolic compounds owing to the increase in the
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surface area and disruption of cell walls present in the plant material used. Usually pure organic
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solvents (such as methanol, ethanol, acetone and ethyl acetate) as well as their combinations with

water (depending on the selectivity, price and recover) are used for extracting phenolic compounds.
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HPLC-DAD (high performance liquid chromatography-diode array detector) is generally used for

elucidating the profiles of phenolic compounds present in many plant species such as legumes. In

contrast to the spectrophotometric techniques (such as Folin-Ciocalteu phenol reagent) this method is

ideal not only for separating phenolic compounds but also their quantification (Khoddami et al.,

2013; Talukdar, 2013). Moreover, despite being economic, simple and quick spectrophotometric

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techniques are not selective and do not quantify individual phenolic compounds (Khoddami et al.,

2013; Magalhães et al., 2017). Generally mass spectrometry is used for the validation of the phenolic

compounds present in various plants such as legume seeds.

Food legumes mainly contain phenolic acids, flavonoids and condensed tannins among

various known phenolic compounds (Amarowicz, and Pegg 2008). These compounds are distributed

differently in the seed coat (mainly flavonoids) and the cotyledon (mainly contain non-flavonoids

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such hydroxycinnamic and hydroxybenzoic acids) (Shahidi and Ambigaipalan, 2015). These

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differences have been highlighted in many reports. Colored (Pinto) beans have more phenolic

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compounds than uncolored (Cannellini) beans (Aguilera et al., 2011). The phenolic compounds

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present in grain legumes are shown in Table 1. For any specific legume, more pigmented varieties

tend to contain higher levels of phenolic compounds (such as anthocyanins) than less pigmented
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varieties (Duenas et al., 2006; Choung et al., 2003). Aguilera et al. (2010) reported 35 phenolic

compounds in lentils including catechins as well as procyanidins (69%), flavonols (17%), flavones,
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flavanones (5%), hydroxybenzoic (5%) and hydroxycinnamic acids (4%). Luthria and Pastor-
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Corrales (2006) separated as well as quantified phenolic acids in 15 dry bean varieties using HPLC.

These authors identified p-coumaric acid, sinapic acid and ferulic acid (most abundant) in all
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varieties, while caffeic acid was identified only in two varieties of black beans.
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2.1. Phenolic acids

The phenolic acids include hydroxybenzoic acid derivatives such as p-hydroxybenzoic,


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protocatechuic, gallic, vanillic and syringic acids and hydroxycinnamic acid derivatives such as

caffeic, ferulic, p-coumaric and sinapic acids.

2.1.1. Hydroxybenzoic acids

Hydroxybenzoic acids have a C6–C1 structure and include gallic, vanillic, protocatechuic, p-

hydroxybenzoic and syringic acid. The structure of hydroxybenzoic acids in grain legume seeds is

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shown in Figure 1a. Beans, peas and lentils varies in concentrations of vanillic, p-hydroxybenzoic

and protocatechuic acids (López-Amorós et al., 2006). Raw lentils had total 5.69 μg/g of

hydroxybenzoics and it included dihydroxybenzoic acid, vanillic acid glycoside, protocatechuic

aldehyde and p-hydroxybenzoic acid (Aguilera et al., 2010). Dihydroxybenzoic acid (3.68 μg/g) and

p-hydroxybenzoic acid (1.90 μg/g) were common phenolic acids in lentils. An early study by

Bartolomé et al. (1997) reported p-hydroxybenzoic acid (1.48 μg/g), protocatechuic acid (0.36 μg/g)

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and protocatechuic aldehyde (0.13 μg/g) in lentils. Alshikh et al. (2015) identified gallic acid, methyl

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vanillate and protocatechuic acid in lentil cultivars. p-hydroxybenzoic (3.25 μg/g) and protocatechuic

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acid (1.45 μg/g) were reported in raw lentils (Dueñas et al 2007). Gallic acid (90.9-136.8 μg/g),

μg/g), μg/g),

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protocatechuic acid (20.2-37.7 2,3,4-trihydroxybenzoic acid (16.9-29.2

protocatechualdehyde (3.6-12.1 μg/g), p-hydroxybenzoic acid (15.7-44.9 μg/g) and vanillic acid
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(0.59-3.22 μg/g) are the benzoic type phenolic acids detected in 11 lentil cultivars (Xu and Chang,

2010).
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Cannellini (Uncolored) and Pinto (Colored) bean had 21.66 and 84.92 μg/g of hydroxybenzoics,
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respectively (Aguilera et al., 2011). This study identified salicylic acid (44.89μg/g), vanillic acid

(17.01 μg/g), p-hydroxybenzoic acid (12.20 μg/g), p-hydroxyphenyl acetic acid (8.42 μg/g) and
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protocatechuic acid (2.40 μg/g) in Pinto beans, whereas vanillic acid (10.71 μg/g), p-hydroxyphenyl
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acetic acid (6.92 μg/g) and p-hydroxybenzoic acid (4.30 μg/g) were the reported hydroxybenzoics in

Cannellini beans. The seed coat (hull) of legume seeds primarily contains p-hydroxybenzoic,
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protocatechuic, gallic, vanillic and syringic acid (Amarowicz, and Pegg 2008). Protocatechuic acid

(67.6 μg/g) and protocatechuic aldehyde (7.71 μg/g) were identified in crude extract of adzuki bean

(Amarowicz et al., 2008), whereas gallic aldehyde (13.45 μg/g) and p-hydroxybenzoic acid (73.46

μg/g) were reported in red lentil acetone extract (Amarowicz et al., 2009). Gallic (27 μg/g),

protocatechuic (18.9 μg/g) and p-hydroxybenzoic (5.81μg/g) are the main free phenolic acids

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identified in black cowpea seed coats (Gutiérrez-Uribe et al., 2011). Protocatechuic (0.217 mg/g) and

p-hydroxybenzoic acid (0.239 mg/g) were present in alkaline hydrolysable phenolic fractions of dark

colored cranberry beans seed coats, while were not observed in similar fraction of light colored

cranberry beans seed coats (Chen et al., 2015a).

Gallic and protocatechuic are the common phenolic acids found in beans (Wang et al., 2016). Red

sword bean and black sword bean coats contains 9.68 and 5.0 mg/g DW (dry weight basis) of gallic

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acid, respectively (Gan et al., 2016). p-hydroxybenzoic acid (29.08 to 71.26 μg/g) was detected as

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main hydroxylbenzoic in alkaline hydrolysable phenolic fractions of regular darkening cultivars of

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whole cranberry beans (Chen et al., 2015b). p-hydroxybenzoic acid (19.2 to 60.5 mg/kg), syringic

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acid (45.9 mg/kg) and gentisic acid (8.1 to 26.0 mg/kg) were reported in seeds of six chickpea

varieties, while protocatechuic acid (12.1 to 163.5 mg/kg) and p-hydroxybenzoic acid (45.4 to 101.7
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mg/kg) were present in seeds of six field pea varieties (Magalhães et al., 2017). Benzoic (57 μg/g)

and protocatechuic (44 μg/g) acids are main phenolic acids in soybean seed (Chung et al., 2011). p-
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hydroxybenzoic acid was found in kidney beans and its sprouts at level of 10.33 and 10 μg/g,
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respectively (Dueñas et al 2015). Dark beans have 7.3 and 20.4 μg/g of total hydroxybenzoics

(protocatechuic acid, protocatechuic aldehyde, p-hydroxybenzoic acid, p-hydroxybenzoic aldehyde)


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in soluble and insoluble dietary fiber, respectively (Dueñas et al 2016). Study also reported raw lentils
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have high level of hydroxybenzoics in insoluble (73.2 μg/g) than soluble (2 μg/g) dietary fiber.

2.1.2. Hydroxycinnamic acids


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Hydroxycinnamic acids present in grain legume seeds is shown in Figure 1b. They are

aromatic compounds having a C6–C3 structure and include caffeic, p-coumaric, trans-ferulic, sinapic

and chlorogenic acids. Hydroxycinnamics vary in legumes such as beans, peas and lentils. Trans-

ferulic acid is detected in beans, peas and lentils and cis-ferulic acid only in beans. trans-p-coumaric

acid is detected in lentils and peas, and cis-p-coumaric acid only in peas (López-Amorós et al., 2006).

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In 15 dry edible bean varieties, mean ferulic, p-coumaric and sinapic acid was determined as 17.8, 6.3

and 7.0 mg/100 g, respectively (Luthria and Pastor-Corrales, 2006). Lentils contained 3.76 μg/g of

hydroxycinnamics and it included trans-p-coumaric, cis-p-coumaric and hydroxy acids (Aguilera et

al., 2010). Raw lentils contain trans-p-coumaroyl malic acid (10.02 μg/g), trans-p-coumaroyl glycolic

acid (2.88 μg/g), trans-p-coumaric acid (5.74 μg/g) as main hydroxycimmamic compounds (Dueñas

et al 2007). Sinapic (1099-2217 μg/g) and chlorogenic acid (159-213 μg/g) are the main cinnamic

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type phenolic acids detected in 11 lentil cultivars (Xu and Chang, 2010).

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Cannellini and Pinto bean had 23.52 and 36.31 μg/g of hydroxycinnamic acids, respectively with

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majority of them being the aldaric derivatives of ferulic and p-coumaric free acids (Aguilera et al.,

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2011). Further, the main hydroxycinnamic acid identified in Cannellini and Pinto beans was trans-

ferulic acid (8.95 and 11.80 μg/g, respectively). Caffeic, ferulic, p-coumaric and sinapic acids were
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detected in different lentil cultivars (Alshikh et al., 2015). In another study, trans-ferulic, trans-p-

coumaric and sinapic acids were identified in red lentils (Amarowicz et al., 2009). Bartolome et al.
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(1997) identified p-coumaric acid (7.51 μg/g) and p-coumaric acid derivative (0.74 μg/g) in raw
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lentils. Hull portions of legume seeds had trans-p-coumaric, trans-ferulic, ferulic, caffeic and sinapic

acids (Amarowicz, and Pegg 2008). In crude extract of adzuki bean trans-p-coumaric acid (31.3μg/g)
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and trans-p-coumaroyl malic acid (4.57 μg/g) was identified (Amarowicz et al. 2008), whereas in red
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lentil extract, trans-p-coumaric acid (38.84 μg/g) and sinapic acid (0.06 μg/g) was reported

(Amarowicz et al., 2009). Trans-p- coumaric acid (37.3 μg/g), trans-p- coumaric acid derivative (6.4
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μg/g) and trans-ferulic acid (10.1 μg/g) were reported in green lentil (Amarowicz et al. 2010). Ferulic

(95 μg/g) is a main hydroxycinnamic acid in soybean seed (Chung et al., 2011). Ferulic (26.25 μg/g)

and coumaric (1.25 μg/g) are the free phenolic acids found in black cowpea seeds (Gutiérrez-Uribe et

al., 2011). Aldaric derivatives of ferulic, p-coumaric and sinapic acids are main phenolic acids

identified in white kidney bean and round purple bean (García-Lafuente et al., 2014).

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Hydroxycinnamic acids in free forms and linked to hydroxyl acids and sugars are abundant in kidney

beans and they account for nearly about 50% of the total phenolic content (Dueñas et al 2015). Dark

beans have 1.2 and 1.3 μg/g of hydroxycinnamic compounds in soluble and insoluble dietary

fractions, respectively (Dueñas et al 2016). Ferulic and p-coumaric are the two main

hydroxycinnamic acids commonly found in selected beans (Wang et al., 2016).

2.1.3. Total phenolic content

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Total phenolic content (TPC) reported in different legume seeds are presented in Table 1. The

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antioxidant activity of legumes is directly related to TPC (Xu et al., 2007). TPC shows a wide

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variability in different legumes and depends on the source of legume seeds and the region of

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cultivation (Amarowicz, and Pegg 2008). Xu et al. (2007) reported that lentils contain more TPC

(4.86–9.60 mg GAE [gallic acid equivalents]/g) than soybeans (1.57- 5.57 mg GAE/g), chickpeas
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(0.98 mg GAE/g), yellow peas (0.85-1.14 mg GAE/g) and green peas (0.65-0.99 mg GAE/g). Study

also reported that TPC varies widely among common bean varieties (0.57-6.99 mg GAE/g) with
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highest in black turtle beans and lowest in navy beans. The pigmented seeds of legumes contain
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higher TPC than the light-colored seeds. The seed coat of legumes is rich in phenolic compounds,

although it represents approximately 10% of the total seed weight. Segev et al. (2010) documented
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that colored chickpea seeds had up to 13 fold more TPC than beige as well as regular cream colored
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seeds indicating that these legumes can be used as functional foods. TPC in regular-darkening

cranberry bean seed coats is approximately 25-fold higher (52.41–56.43 mg GAE/g) than non-
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darkening (2.00–2.06 mg GAE/g) cranberry beans (Chen et al., 2015a). TPC in raw non-darkening

cranberry beans is 5-fold lower (0.67 – 0.81 mg GAE/g) compared to raw regular-darkening (2.82 –

4.15 mg GAE/g) cranberry beans (Chen et al., 2015b). TPC ranging from 11.2 to 25.3 mg catechin

equivalents (CE)/g was reported for cotyledons of 7 cultivars of beans (Rocha-Guzmán et al. 2007).

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In lentils, TPC in free, esterified and insoluble bound forms was in the range of 1.37–5.53, 2.32–

21.54, 2.55–17.51 mg GAE/g, respectively (Alshikh et al., 2015).

TPC in horse gram, cowpea and chickpea flour extracts was reported to be 14.3, 12.1 and 10.8 mg

GAE/g, respectively (Sreerama et al., 2012). TPC varies from cultivar to cultivar in lentils and this

might be due to difference in geographical conditions and locations of the cultivars (Alshikh et al.,

2015). TPC in legume seeds such as adzuki bean, faba bean, red bean, broad bean, pea, red lentil and

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green lentil was greatly diversified and reported as 89.7, 55.9, 55.4, 23.9, 22.6, 58.0 and 67.6 mg/g

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catechin equivalent (CE), respectively (Amarowicz et al., 2004). TPC in crude extract, low-

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molecular-weight phenolic fraction and tannin fraction of adzuki bean was reported to be 90, 58 and

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189 mg/g (Amarowicz et al., 2008), whereas in red lentil it was 58, 12 and 290 mg/g, respectively

(Amarowicz et al., 2009). TPC values were higher (68, 23 and 338 mg/g, respectively) in similar
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extract and fractions of green lentil (Amarowicz et al., 2010). TPC in seed coat and cotyledons of

thirteen genotypes of faba bean was reported in the range of 45.6–107.6 and 15.8–26.3 mg GAE/g,
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respectively (Chaieb et al., 2011).Seed coat contributed to the majority of phenolic compounds in
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peanuts. TPC in seed coat, raw kernel and cotyledons of six peanut varieties ranged from 97.3 to

133.5, 3.28–9.20 and 0.88–1.85 mg GAE/g, respectively (Attree et al., 2015). TPC in immature seeds
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of ten varieties of faba beans ranged from 817 to 1337 mg GAE/kg (Baginsky et al., 2013). Red and
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carioca beans presented higher TPC (1.8 and 1.2 mg/g, respectively) compared to fradinho, white and

black beans (0.18; 0.19 and 0.49 mg/g respectively) (Telles et al., 2017).
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2.2. Flavonoids

Flavonoids are the main phenolic compounds found in legumes (Aguilera et al., 2011). They

are the largest group of phenolics and account for the half of known phenolic compounds in plants.

They are low molecular weight compounds consisting of two aromatic rings (A & B) joined by a 3-

carbon bridge (C6–C3–C6 structure). The ring A is derived from the acetate pathway and ring B is

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derived from shikimic acid pathway. They are divided into anthocyanins (colored) and anthoxanthins

(colorless compounds). Anthoxanthins are divided further into flavones, flavans, flavonols,

isoflavones and their glycosides. Flavanols, flavan-3-ols, flavones and anthocyanidins are the main

flavonoids present in leguminous seeds (Amarowicz and Pegg, 2008).

2.2.1. Catechins and proanthocyanidins

Catechins have a hydroxyl group at C-3 and are called as flavan-3-ols. Procyanidins are the

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oligomers of catechin and epicatechin molecules. Catechins and procyanidins identified in grain

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legume seeds are shown in Figure 2a. These flavonoids are primarily present in legumes having

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colored seed coats. Aguilera et al. (2011) reported that colored (Pinto) beans contained 256.94 μg/g of

catechins and procyanidins and the most abundant compounds were (+)-catechin (142.58 μg/g), (+)-

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catechin glucoside (45.75 μg/g) and procyanidin dimer 1 (41.20 μg/g). Catechins, procyanidins and
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prodelphinidins are common in raw lentil flour sample (Dueñas et al 2007). Catechins and

procyanidins represent 69% of total phenolic compounds (74.48 μg/g) in raw lentils (Aguilera et al.,
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2010). Catechin (267-1899 μg/g) and epicatechin (2535-4946 μg/g) are the two major
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compounds detected in 11 lentil cultivars (Xu and Chang, 2010). The mainly identified

compounds in lentils were (+)-catechin 3-glucoside (39.89 μg/g), procyanidin B2 (8.92 μg/g),
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procyanidin trimer (9.30 μg/g) and procyanidin dimer (4.45 μg/g) (Aguilera et al., 2010). Catechin,
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epicatechin (+)-catechin 3-glucoside were detected as catechins and procyanidin dimer A & B,

prodelphinidin dimer A and procyanidin trimer C1 & A as procyanidins in lentil cultivars (Alshikh et
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al., 2015). Insoluble dietary fractions of raw lentils contain catechins (1.37 μg/g) and procyandins

(1.03 μg/g), whereas they are not detected in soluble dietary fractions (Dueñas et al 2016). Catechin

glucoside (688 μg/g), epicatechin (25.7 μg/g), epicatechin glucoside (159 μg/g), epigallocatechin

gallate (0.14 μg/g) were detected in crude extract of adzuki bean (Amarowicz et al., 2008). In red

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lentil crude extract, epicatechin (98.21 μg/g), catechin (36.02 μg/g), catechin glucoside (51.95 μg/g),

and epicatechin glucoside (6.65 μg/g) were measured (Amarowicz et al., 2009).

Hull of colored beans (red, brown, and black bean) contain procyanidin B2, C1, and C2

(Amarowicz, and Pegg 2008). Catechin is the most predominant flavonoid in dark colored seed coat

of cranberry beans and its levels is 2.5 to 6.5-fold higher than proanthocyanidins. In free phenolic and

alkaline hydrolysable phenolic fractions of regular-darkening cranberry beans seed coats the level of

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catechin is 3.7 and 0.25 mg/g and procyanidin dimers is 1.8 and 0.14 mg/g, respectively (Chen et al.,

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2015a). Catechins and proanthocyanidins are the main compounds identified in free phenol fraction

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of regular darkening cranberry bean lines. Catechin and proanthocyanidins concentration in whole

raw cranberry beans (regular darkening lines) ranged from 142.26 to 203 and 15.30 to 35.29 μg/g,

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respectively (Chen et al., 2015b). Catechin (1.75–5.42%) and epicatechin (3.80–12.48%) were
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quantified in seed coats of 15 bean cultivars (Mojica et al., 2015). Catechin gallate (2060 μg/g),

digallate procyanidin dimer (864 μg/g), epicatechin (864 μg/g) and gallate procyanidin dimer (676
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μg/g) were the major phenolics present in the broad bean crude extract (Amarowicz and Shahidi,
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2017). Catechin glucoside is the main dominant compound found in cowpeas. Catechin (16.6 - 297

μg/g), catechin-7-O-glucoside (770 - 2553 μg/g), epicatechin (2.4 - 11.3 μg/g) and (epi)afzelechin
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glycosides (10.7 - 243 μg/g) were quantified in six cowpea phenotypes (Ojwang et al., 2013).
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In adzuki bean, the content of procyanidin gallate (12.4 μg/g), procyanidin dimers (16.0-213

μg/g) and procyanidin trimers (41.8-42.4 μg/g) was reported in crude extract (Amarowicz et al.,
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2008). In lentils, the content of procyanidins (procyanidin B2, B3 and procyanidin tetramer) was in

the range of 0.1 to 0.5 mg/100 g DW (López-Amorós et al., 2006). Prodelphinidin dimers (161.6

μg/g), procyanidin dimers (65.98 μg/g), procyanidin trimers (87.5 μg/g), digallate procyanidin dimer

(83.29 μg/g), procyanidin gallate (32.78 μg/g) were reported in red lentil acetone extract (Amarowicz

et al., 2009). Prodelphinidin dimer (5.3 μg/g), digallate procyanidin dimer (7.3 μg/g), procyanidin

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dimers 1 (14.1 μg/g) & 2 (100 μg/g), procyanidin dimer gallate (31.9 μg/g), digallate procyanidin

dimer (14.1 μg/g), procyanidin trimer (18.6 μg/g), catechin glucoside (289 μg/g), catechin gallate

(18.7 μg/g), epicatechin glucoside (59.5 μg/g) were reported in green lentil (Amarowicz et al., 2010).

Concentration of proanthocyanidins detected in immature seeds of ten faba bean varieties ranged

from 543 to 36.55 mg/kg fresh weight (Baginsky et al., 2013). Catechin (151-978 mg/kg), epicatechin

(148-773 mg/kg), procyanidin dimer (71-1352 mg/kg), procyanidin trimer (64-782 mg/kg) and

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prodelphinidin dimer (81-1150 mg/kg) were the main compounds identified in seeds of ten faba bean

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varieties (Baginsky et al., 2013). The high concentration of procyanidin dimers (97.29 μg/g) were

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reported in raw dark beans (López et al., 2013).

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2.2.2. Anthocyanins

Anthocyanidins (or aglycons) are the main structures of anthocyanins and consist of an
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aromatic ring A that is bonded to a heterocyclic ring C which contains oxygen which forms a bond

(carbon–carbon) with the third aromatic ring B. When anthocyanidins are present in their glycoside
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form i.e. bonded to a sugar moiety, these compounds are referred to as anthocyanins. Anthocyanins
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are mostly present in the colored seed coat of the legumes. Structure of main anthocyanins identified

in grain legume seeds is shown in Figure 2b. In red and green lentils, anthocyanin contents reported is
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three to four folds higher in hulls than those in whole seeds (Oomah et al., 2011). Anthocyanin
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content of 157.3 and 665.6 μg/g was reported in French Green and Pardina lentils, respectively (Xu

and Chang, 2010). In same study, six folds higher anthocyanin content was reported in yellow pea
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hull than those in whole seed. Cyanidin 3-glucoside (88.44 μg/g) and pelargonidin 3-glucoside (50.72

μg/g) are the most abundant anthocyanins identified in raw dark beans (López et al., 2013).

Monomeric anthocyanin content (MAC) was higher in black seed coat (73.7 mg cyanidin-3-

glucoside equivalents (CyE) /g) compared to red seed coat (9.06 mg CyE/g) of peanuts (Attree et al.

2015). Malvidin, pelargonidin and cyanidin glucosides identified in round purple bean extract are the

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anthocyanins responsible for color of this bean (García-Lafuente et al., 2014). Pelargonidin 3-O-

glucoside (1.90 μg/g), cyanidin-3-O-glucoside (0.31 μg/g) and malvidin-3-O-glucoside (0.06 μg/g)

are the main anthocyanins in kidney bean (Dueñas et al 2015). Delphinidin (0.9–129.0 mg/100 g),

petunidin (0.7–115.0 mg/100 g) and malvidin (0.14–52.0 mg/100 g) glucosides are the three

anthocyamins quantified in seed coats of 15 bean cultivars from Mexico and Brazil (Mojica et al.,

2015). Study reported that seed coats of two black bean cultivars have the highest concentration of

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total anthocyanins (206 & 250 mg/100 g). The contents of cyanidin, pelargonidin, petunidin,

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malvidin, delphinidin and the total anthocyanindin in 26 kidney bean varieties ranged from 0–1.44,

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0–0.71, 0–0.41, 0–0.27, 0–4.45 and 0–5.84 mg/g bean coat DW, respectively (Kan et al., 2016a).

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Han et al. (2015) reported that anthocyanins were responsible for the pigmentation of the adzuki bean

extracts. They identified malvidin-3-O-glucoside and peonidin-3-rutinoside as two new anthocyanin


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compounds in the adzuki beans.

2.2.3. Flavonols and flavones


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The structures of flavonols (myricetin, quercetin and kaempferol) and flavones (apigenin and
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luteolin) identified in legume seeds is shown in Figure 3a and 3b, respectively. Myricetin 3-O-

rhamnoside (5.95 μg/g) and quercetin 3-O-rutinoside (5.24 μg/g) are the main flavonol glycosides
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identified in raw lentils. Apigenin 7-O-apiofuranosyl glucoside (6.20 μg/g), apigenin 7-O-glucoside
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(1.87 μg/g), and luteolin 7-O-glucoside (1.29 μg/g), are the main flavone glycosides identified in raw

lentils (Dueñas et al 2007). Seventeen percent of the total identified phenols in raw lentil are
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flavonols and dihydroflavonols (Aguilera et al., 2010). Kaempferol glycosides were the main

flavonols identified in raw lentils and the most common were kaempferol 3-rutinoside (5.95μg/g) and

kaempferol glucoside (3.66 μg/g). Alshikh et al. (2015) identified kaempferol dirutinoside as flavonol

in six lentil cultivars. Aguilera et al. (2011) reported that 4% of identified phenols in raw Pinto beans

were flavonols and these included quercetin glucoside (7.15 μg/g) and kaempferol glucoside (3.13

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μg/g) as main components. Quercetin (36.2 μg/g), quercetin glucoside (181 μg/g), quercetin

galactoside (46.9 μg/g), quercetin arabinglucoside (42.8 μg/g), quercetin rutinoside (38.2 μg/g),

dihydroquercetin (1.15 μg/g), kaempferol rutinoside (38.2 μg/g) and myricetin rhamnoside (212

μg/g) were identified in crude extract of adzuki bean (Amarowicz et al., 2008).

In red lentil acetone extract, kaempferol derivative (37.56 μg/g) and quercetin hexose acylated

(21.44 μg/g) were identified (Amarowicz et al., 2009). In crude extract of green lentil quercetin

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diglycoside (114 μg/g) was the dominant compound and other flavonols identified were quercetin

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hexose acylated (37.2 μg/g) and kaempferol glucoside(19.4 μg/g) (Amarowicz et al., 2010). Luteolin

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in the range from 21.8 to 77.1 μg/g was quantified in 11 lentil cultivars (Xu and Chang, 2010).

Luteolin 3´-7-diglucoside (4.55μg/g) was the main flavone identified in lentils (Aguilera et al., 2010).

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Alshikh et al. (2015) also reported presence of the same flavone in lentils. Kaempferol (6.09 μg/g),
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quercetin (4.02 μg/g), and myricetin (2.30 μg/g) are the free flavonols present in black cowpea seed

coats (Gutiérrez-Uribe et al. (2011). Mirali et al. (2014) used HPLC-MS method for identifying
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phenolic compounds present in the seed coats (black, green and grey) of 3 lentil genotypes. The
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contents of oligomeric flavan-3-ols were almost similar in all three genotypes, whereas luteolin

(including its glycosylated form) was more abundant in the genotype containing black seed coat.
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Total flavonols and flavones detected in immature seeds of ten faba bean varieties ranged from 101 to
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377 mg/kg fresh weight (Baginsky et al., 2013). Myricetin (18.8-119.5 mg/kg), myricetin glucoside

(9.9-41.9 mg/kg), quercetin galactoside (8.3-92.4 mg/kg), quercetin rutinoside (7.4-56.9 mg/kg),
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quercetin glucoside (5.1-25.9 mg/kg) and apigenin galactoside (8.9-168 mg/kg) were identified in ten

faba bean varieties. Quercetin and its derivatives are the flavonols present only in colored (round

purple) beans and they were absent in white kidney beans (García-Lafuente et al., 2014).

Most of the flavonols and flavones are concentrated in insoluble dietary fiber fractions of dark

beans and lentils (Dueñas et al 2016). Dark beans contain kaempferol dihexoside-rhamnoside

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(1.06μg/g) and lentils contain kaempferol (1.64 μg/g), kaempferol dihexoside (1.37μg/g), kaempferol

rutinoside (0.30μg/g), kaempferol-3-O-glucoside (0.61 μg/g), quercetin-3-O-glucoside (1.0 μg/g) and

luteolin rhamnoside-hexoside (0.36 μg/g) in insoluble dietary fiber fractions. Myricetin-3-O-

rhamnoside (7.4 mg/kg), luteolin-8-C-glucoside (1.2 mg/kg), quercetin-3-O-rhamnoside (5.0 mg/kg)

and quercetin-3-O-galactoside (7.2) were reported in Elmo chickpea variety (Magalhães et al., 2017).

In mung beans coats, apigenin-8-C-glucoside (vitexin) and apigenin-6-C-glucoside (isovitexin) were

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quantified in the range from 3.31 to 3.53 and 4.83 to 7.13 mg/g DW, respectively (Gen et al., 2016).

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Apigenin-8-C-glucoside (8.5 to 21.2 mg/kg), apigenin-6-C-glucoside (2.7 to 24.6 mg/kg) and

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luteolin-6-C-glucoside (1.4 to 11.3 mg/kg) were reported in field pea varieties (Magalhães et al.,

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2017).

2.2.4. Flavanones
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The flavanones identified in legume seeds are pinocembrin, naringenin, eriodictyol,

hesperitin, and sakuranetin (Figure 3c). The total flavanones present in Pinto and Cannellini beans
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were 10.87 and 7.06 μg/g, respectively (Aguilera et al., 2011). Sakuranetin (4.49 μg/g), eriodictyol
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(3.83 μg/g), naringenin derivative (1.29 μg/g) and pinocembrin (1.26 μg/g) were reported in Pinto

beans. Eriodictyol derivative (2.01 μg/g), sakuranetin (1.59 μg/g), pinocembrin derivative 2 (1.28
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μg/g) and eriodictyol (1.20 μg/g) were the identified flavanones in Cannellini beans (Aguilera et al.,
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2011). Naringenin (11.30 μg/g) and hesperitin (0.14 μg/g) are the flavanon-glycoside reported in

white kidney bean (García-Lafuente et al., 2014).


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2.2.5. Total flavonoid content

Total flavonoid content (TFC) in legumes such as chickpea, green pea, yellow pea, red

kidney, lentils and black bean depend upon growing location and post-harvest storage conditions. The

TFC reported in various legumes is shown in Table 2. The TFC of lentils ranged from 3.04 to 4.54

mg CE/g; common beans from 0.92 to 4.24 mg CE/g; yellow pea from 0.09 to 0.17 mg CE/g; green

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pea from 0.05 to 0.15 mg CE/g and soybean from 1.06 to 4.04 mg CE/g (Xu et al., 2007). Black and

red chickpea seeds contain 10 to 11 times more TFC than beige and regular cream colored chickpea

seeds (Segev et al., 2010). TFC in the range of 2840 to 6870 μg/g was reported in 11 lentil

cultivars (Xu and Chang, 2010). Horse gram, cowpea and chickpea flour extracts contains TFC of

8.6, 7.2 and 4.8 mg CE/g, respectively (Sreerama et al., 2012). 3345 μg/g flavanoids content is

reported in the soybean seed (Chung et al., 2011). Seed coats are the main contributors of the

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flavonoids in peanuts. TFC in seed coat, raw kernel and cotyledons of six peanut varieties was

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reported in the range of 31.68 to 85.17, 0.97 to 2.84 and 0.18–0.30 mg CE/g, respectively (Attree et

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al., 2015). In black cowpea seed coats, 10 times more flavonoids were reported compared to whole

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seeds whereas cowpea cotyledons are devoid of these compounds (Gutiérrez-Uribe et al.,2011). TFC

in seed coat and cotyledons of thirteen genotypes of faba bean ranged from 5.3–17.5 and 3.7–5.9 mg
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rutin equivalents (RE)/g sample, respectively (Chaieb et al., 2011). TFC in free, esterified and

insoluble bound forms in lentils was in the range of 0.01–0.80, 0.36–4.13 and 0.08-2.95 mg CE/g,
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respectively (Alshikh et al., 2015).


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2.3. Tannins

Tannins are the polyphenols having molecular weight ranging from 500 to 3000 D found in
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complexes with polysaccharides, alkaloids and proteins. These compounds can be classified as
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hydrolysable and non-hydrolysable tannins (condensed tannins). Tannins are primarily present in the

testa of legumes, playing important roles in the defense system of these seeds which normally get
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exposed to oxidative damage (from various environmental factors) (Shahidi and Ambigaipalan,

2015). Legumes with colored seed coats such as lentils, red beans and black beans contain high

amounts of condensed tannins (Amarowicz, and Pegg 2008). Condensed tannins (proanthocyanidins)

are oligomeric and polymeric flavonoids (flavan-3-ols) that release anthocyanidins and catechins by

heat treatment in alcoholic solutions. Condensed tannins of lentil, pea and faba beans consist of

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catechin, gallocatechin, epigallocatechin and epicatechin subunits (Jin et al., 2012). Among peas,

proanthocyanidin subunit composition was primarily prodelphinidin (epigallocatechin and

gallocatechin), whereas in lentils and faba beans it was prodelphinidin and procyanidin-type flavan-3-

ol subunits. Glycosides of catechin and (epi)afzelechin are the major subunits in proanthocyanidins

of cowpea with catechin-7-O-glucoside as predominant flavan-3-ols accounted for 88% of the

monomers (Ojwang et al., 2013). Catechin, epicatechin, gallocatechin and epigallocatechin are the

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subunits in oligomers of proanthocyanidins detected in lentil seed coat (Mirali et al., 2014). Studies

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proved that tannins vary in subunit composition among legume seeds.

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2.3.1. Condensed tannin content

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There is great variation in level of condensed tannin content (CTC) among commonly

consumed legumes (Table 3). CTC of lentil cultivars ranged from 3.73 to 10.20 mg CE/g; common
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beans varieties from 0.47 to 5.73 mg CE/g; soyabean cultivars from 1.06 to 4.04 mg CE/g; yellow pea

cultivars ranged from 0.22 to 0.59 mg CE/g and green pea cultivars from 0.23 to 0.61 mg CE/g (Xu et
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al., 2007). Legumes having colored or pigmented seed coats are natural source of condensed tannins.
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Seed coats of peanuts have high level of condensed tannins compared to kernel and cotyledons

(Attree et al., 2015). CTC in seed coat, raw kernel and cotyledons of six peanut varieties was reported
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in the range of 29.7 to 84.7, 2.88 to 4.73 and 1.74 to 3.75 mg CE/g, respectively. The highest levels of
CE

condensed tannins were found in lentils (Alshikh et al., 2015). CTC in free, esterified and insoluble

bound forms in lentils were in the range of 0.40–2.67, 0.64–4.20 and 0.03-5.57 mg CE/g,
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respectively.

Bean hulls had more CTC than whole bean (Madhujith et al., 2004). Tannins content determined

using vanillin method (expressed as absorbance at 500 nm/mg) in faba bean, broad bean, adzuki bean,

red bean, pea, red lentil and green lentil was 0.416, 0.093, 0.483, 0.434, 0.071, 0.577 and 0.545,

respectively (Amarowicz et al., 2004). The tannins content in crude extract, low-molecular-weight

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phenolic fraction and tannin fraction of adzuki bean was reported to be 136, 45 and 213 absorbance

units (Amarowicz et al., 2008), whereas in red lentil it was reported to be 70, 1.52 and 1.29

absorbance units at 500 nm/g, respectively (Amarowicz et al., 2009). Green lentils had more tannin

content as the values for crude extract and two fractions was obtained as 93, 1.92 and 252 A500/g

(Amarowicz et al., 2010).

CTC ranged from 309 to 958 mg CE/kg in immature seeds of ten bean varieties (Baginsky et al.,

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2013). Condensed tannins are the main polyphenols reported in pigmented bean coats (Gan et al.,

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2016). The mean proanthocyanidin (PA) content reported in faba bean, pea and lentil is 654.3, 33.6

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and 329.4 mg/100 g fresh weight, respectively (Jin et al., 2012). Chickpea flour had the highest PA

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content (5.1 mg CE/g) than horse gram (3.8 mg CE/g) and cowpea (2.1 mg CE/g) flours (Sreerama et

al., 2012). Six cowpea phenotypes with different seed coat color (black, red, green, white, light-
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brown and golden-brown) contains PA content in the range from 2.2 to 6.3 mg/g DW (Ojwang et al.,

2013). Regular raw dark colored cranberry beans have 60- fold higher PA contents compared to raw
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light colored cranberry beans (Chen et al., 2015b). PA content varied widely in 26 kidney beans
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cultivars due to diversity in color of seed coat and it was observed in range of 10.52 to 57.13 mg

procyanidin B-2 equivalents/gram (Kan et al., 2016a). Moreover, proanthocyanidins were not
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detected in kidney bean cultivars with white color seed coats.


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3. Effect of processing

Phenolic compounds present in legumes are of nutritional interest. The changes in level of
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phenolic compounds during processing might modify the nutritional value of legumes. Milling,

dehulling, soaking, germination, cooking and fermentation are common in legumes before human

consumption. Germination process modifies phenolic composition of beans, peas and lentils

depending upon the germination conditions (López-Amorós et al., 2006). Lentils showed decrease in

antioxidant activity and functionality whereas peas and beans showed increase under similar

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germination conditions. Bartolomé et al. (1997) reported that germination does not change the level

of phenolic compounds, whereas fermentation increased the content of some individual phenolic

compounds. Increase in level of protocatechuic acid (1.10 from 0.36μg/g), p-hydroxybenzoic acid

(7.39 from 1.48μg/g) and catechin (17.53 from 7.31μg/g) was reported in fermented lentils compared

to raw lentils. TPC reported in fermented chickpea seeds (1.69 mg catechin/g DW) is higher

compared to germinate (0.99 mg catechin/g DW) and raw chickpea (0.54 mg catechin/g DW) seeds

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(Fernandez-Orozco et al., 2009).

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Cooking and germination brings important changes in bioactive compounds of legume seeds.

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Level of phenolic compounds increases during germination and decreases during cooking process

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(López-Martínez et al., 2017). Soaking, boiling and steaming processes significantly affected TPC

and antioxidant activities in green pea, yellow pea, chickpea and lentils (Xu and Chang, 2008).
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Soaking resulted in to loss of 2–12% of TPC in peas and chickpeas and 9–38% in lentils. In this

study, steaming treatments exhibited several advantages compared to boiling treatments and resulted
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into greater retention of TPC and antioxidant activities in legumes. The nutritional value attributed by
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phenolic acids does not changes significantly by overnight soaking of dry beans and 83% of the

phenolic acids were retained in 15 dry bean samples after cooking (Luthria and Pastor-Corrales,
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2006). Recent study reports that cooking and germination improves health-promoting properties of
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beans and lentils by bringing positive changes in phenolic composition (Dueñas et al 2016). TPC

remains stable even after 8 days of germination in kidney beans (Dueñas et al 2015).Study reports
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that germination decreases anthocyanins and flavan-3-ols levels and it was compensated with a higher

content of flavonols and flavanones in germinated kidney beans. Soaking of lentils increased the

dihydroxybenzoic acid level from 3.68 μg/g (raw lentils) to 39.20 μg/g and after cooking as well as

dehydration process lentils contained 31.69 μg/g of this phenolic compound (Aguilera et al., 2010).

Increase in the level of hydroxybenzoic acid might be due to the breakdown of insoluble phenolic

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compounds and disruption of cell wall during processing. Germination of pigeon pea improves TPC

and antioxidant activity by 30% and 63%, respectively (Uchegbu and Ishiwu, 2016).

Legumes with high level of free phenolics and flavonoids in their seed coats are more prone to

lose these compounds during soaking and germination (Gutiérrez-Uribe et al., 2011). TPC and

antioxidant activity decreased by 80% in lentils and 30% in yellow peas by seed coat removal, 22–

42% in lentils by soaking, and 16–41% in lentils, chickpeas and peas by cooking (Han and Baik,

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2008). Soaking of Cannellini and Pinto beans reduced the hydroxybenzoics by 95% and 81%,

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respectively (Aguilera et al., 2011). Dehydration of soaked and cooked Cannellini and Pinto beans

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slightly improved the hydroxybenzoic acid level as compared to soaking and cooking in this study.

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Hydroxycinnamics are lost due to leaching into cooking water or owing to breakdown during the

processing in lentils (Aguilera et al., 2010). Soaking reduced these compounds up to 42.7% whereas
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soaking, cooking and dehydrated exhibited lowest decrease (16.5%) in Cannellini beans (Aguilera et

al., 2011). Other phenolic compounds (catechins and procyanidins, flavonols and dihydroflavonols,
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flavones andflavanones) in lentils were significantly decreased by soaking, cooking and dehydration
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(Aguilera et al., 2010). Catechins and procyanidins were completely lost by dehydration, whereas

soaking showed 85% and soaking along with cooking showed 67% reduction in Pinto bean flours
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(Aguilera et al., 2011).


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Flavonols and dihydroflavonols were drastically reduced (67%) in lentils after processing

(Aguilera et al., 2010). The treatment of lentil flour samples with enzymes (phytase, α-galactosidase,
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viscozyme or tannase) in stirred fermentor decreased level of hydroxycinnamic compounds and

proanthocyanidins (Dueñas et al 2007). Chung et al (2011) reported low percent (47.1%) recovery of

phenolic compounds in tofu due to boiling, leaching in water, coagulation, filtering and whey

exclusion. Thermal processing of lentils releases some of the flavonols from bonded forms so there

level increases as compared to soaked samples. The level of kaempferol dirutinoside, kaempferol

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acetylglycoside, and kaempferol 3-glucoside increased after cooking of soaked samples. Flavones and

flavanones level decreased due to leaching and thermal/oxidative deterioration (Aguilera et al., 2010).

Dry heating, autoclaving and soaking followed by autoclaving affected the TPC and antioxidant

activity in chickpea and pea (Nithiyanantham et al., 2012). In this study, dry heating proved

beneficial as it caused smaller loss of TPC and antioxidant activity in chickpea and pea. Soaking,

cooking and dehydration cause significant decrease in the level of flavonols and flavanones in beans

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(Aguilera et al., 2010). The reduction in phenolic compounds observed during cooking might be due

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to the destruction of these compounds as well as chemical rearrangements such as binding of phenolic

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compounds with other organic materials (López-Martínez et al., 2017). Moreover, this might be due

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to the leaching of water-soluble phenolic compounds into the cooking water as discussed earlier.

However, the conditions of cooking process in which the phenolic compounds are not lost
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significantly can be developed in the future (López-Martínez et al., 2017).

4. Antioxidant activity
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The antioxidant activity of phenolics is directly related to their chemical structures such
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degree of glycosylation and number as well as position of hydroxyl groups attached in relation to the

carboxyl functional group (Aguilera et al., 2011; Balasundram et al. 2006). These compounds make
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important a contribution to antioxidant activity due to their radical scavenging as well as metal
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chelating activity. Free radicals and metal ions have harmful effects on biological systems. Phenolic

compounds have ability to donate hydrogen atom or an electron to free radical to form stable
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intermediates. These compounds scavenge free radicals, decompose oxidation products and chelate

metal ions. Antioxidant activity of phenolic compounds present in edible grain legume seeds is of

interest and commonly reported in many recent studies. The antioxidant activities of various legumes

are shown in Table 4. Total antioxidant activity (TAA) of faba bean, broad bean, adzuki bean, red

bean, pea, red lentil and green lentil extracts was reported to be 0.88, 0.58, 1.76, 1.49, 0.30, 0.68 and

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0.75 µmol trolox/mg, respectively (Amarowicz et al., 2004). The ORAC values of lentil cultivars

ranged from 59.55 to 95.19 μmol TE/g; soybean cultivars from 35.1 to 131.3 μmol TE/g; common

bean varieties from 13.30 to 92.73 μmol of TE/g; yellow pea cultivars from 3.26 to 12.80 μmol TE/g

and green pea cultivars from 1.73 to 9.95 μmol TE/g (Xu et al., 2007).

The values of TAA reported for crude extract, low-molecular-weight phenolic fraction and tannin

fraction of adzuki bean were 1.76, 1.40 and 4.17 µmol trolox/mg (Amarowicz et al., 2008) and for red

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lentil these were 0.68, 0.33 and 5.85 µmol trolox/mg, respectively (Amarowicz et al., 2009).

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Germination and fermentation can further improve antioxidant properties of legume seeds due to

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increase in level of TPC (Fernandez-Orozco et al., 2009). In these studies, it was noticeable that the

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tannin fraction of adzuki bean and red lentil exhibited high antiradical activity compared to the crude

extract and low-molecular-weight phenolic fraction. The high antioxidant potential of some grain
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legume seeds is useful for their utilization in production of healthy foods. Green lentils had TAA

values of 0.75, 0.33 and 6.09 for crude extract and two fractions (Amarowicz et al., 2010). Chickpeas
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with black and red colored seed coats showed 30-31 folds more antioxidant potential than chickpeas
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with beige and cream colored seed coats (Segev et al., 2010). Free radical scavenging ability of horse

gram flour extract is reported to be higher (IC50 22.9 μg/ml) than chickpea (IC50 31.4 μg /ml) and
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cowpea (IC50 48.7 μg/ml) flour extracts (Sreerama et al. (2012). DPPH free radical scavenging
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activity (DPPH assay) for seed coat, raw kernel and cotyledon of six peanut varieties ranged from

25.8 to 28.8, 10.8 to 28.9 and 1.50 to 2.72 µmol trolox equivalents (TE)/g, respectively (Attree et al.,
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2015). The ferric reducing antioxidant power (FRAP) values for seed coats, raw kernels and

cotyledons of six peanut varieties ranged from 626.3 to 1111.3, 19.4 to 62.3 and 2.04 to 4.15 µmol

Fe2+/g, respectively (Attree et al., 2015). Seed coat of peanuts is rich in phenolic compounds and they

contributed 88.2 to 95.8% of the total antioxidant capacity in these peanuts varieties.

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Legumes rich in phenolic compounds showed an interesting level of antioxidant capacity and

they are beneficial for health by the daily inclusion in human diet. Lentils have shown high oxygen

radical absorbing capacity (ORAC) values (66.97 μmol TE/g DW) due to their significant amount of

phenolic compounds (mainly flavonoids) present in the seed coat (Aguilera et al., 2010). Lentil

cultivars with high level of catechin, epicatechin and TFC have high antioxidant properties (Xu and

Chang, 2010). Antioxidant capacity of colored (Pinto) beans was more than uncolored (Cannellini)

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beans and it might be due to the presence of catechins and procyanidins in colored beans (Aguilera et

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al., 2011). The antioxidant activity of regular-darkening seed coats was higher (125.28–287.87 µmol

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TE/g) compared to non-darkening seed coats (2.11–10.09 µmol TE/g) in cranberry beans due to the

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difference in the level of free and conjugated phenolic compounds (Chen et al., 2015a). The ORAC

values of 96.2 and 38.3 μmol trolox/g were reported for Pinto and Cannellini beans, respectively.
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Soaking, cooking and dehydration led to a decrease in antioxidant activity in Pinto and Cannellini

beans and changes were more noticeable in case of Pinto bean owing to the loss of catechins,
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procyanidins and flavonols (Aguilera et al., 2011). Leaching of water-soluble phenols, oxidative
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reaction and solid losses during processing lowers the antioxidant activity of legumes (Aguilera et al.,

2011). Faba beans are a good source of natural antioxidant compounds and their antioxidant activity
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is substantially contributed by phenolic compounds present in seeds. The FRAP values of seed coat
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and cotyledons from 13 faba bean genotypes was reported in the range of 0.273-0.931 and 0.158-

0.766 mmol/g and DPPH scavenging capacity was reported in the range of 19.67- 32.76 and 1.24-
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11.97 %, respectively (Chaieb et al., 2011). Seed coats of 15 bean cultivars showed antioxidant

activity in the range of 185.2 to 233.9 mmol TE/g coat (Mojica et al., 2015). Seed coats of white and

black kidney beans exhibited lower (4.00 µmol TE/g) and higher (491 µmol TE/g) antioxidant values

among 26 tested kidney bean cultivars (Kan et al 2016a). Pigmented (red and black sword) bean coats

have high antioxidant potential and it is much higher than most of the common fruits and vegetables

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(Gan et al., 2016). Flavonoids and proanthocyanidins present in pigmented bean coats contribute

mainly to the antioxidant effect. Wang et al. (2016) determined the TPC and antioxidant activity of

few selected beans originating from China. Antioxidant capacities varied considerably (3.2 to 5.9

µmol trolox/g) in selected beans. A positive correlation was reported between total antioxidant

activities and TPC by these authors.

5. Health benefits

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Phenolic compounds are reactive in nature and are terminators of free radicals as well as

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chelators of metal ions that are capable to catalyze oxidative reactions (Ozcan et al., 2014). The

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health promoting activities of phenolic compounds depend on their structure, conjugation with

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phenolics and other compounds along with solubility, absorption and metabolism (Ozcan et al.,

2014). They affect many physiological activities owing to their antioxidant, antihypertensive and
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antibacterial activity. Phenolic compounds are known for free radicals scavenging, chelation and

stabilization of cations and modulation of antioxidant enzymes (Ozcan et al., 2014). They exert
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modulatory effect on different components of the cells having important role in cellular functions
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such as growth, proliferation and apoptosis (Crozier et al., 2009). Flavonoids, catechins and their

derivatives are used for therapeutic purposes due to their health beneficial properties.
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Phenolic acids and flavonoids are the most widely distributed phenolic compounds in legumes
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seeds with a potent antioxidant activity. The antioxidant activity of phenolic compounds present in

food legumes is of nutritional interest as it is associated with their health promoting effects. Phenolic
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compounds have a role in the growth inhibition of pathogens and decay of microorganisms.

Moreover, phenolic compounds have inhibitory activity against fungal amylase in vitro as determined

in edible beans (Telles et al., 2017). Besides antioxidant activity, phenolic compounds might also

play key roles in inhibition of lipase, -amylase and -glucosidase activities in vitro (Zhang et al.,

2010; Zhang et al., 2015; Tan et al., 2017). Inhibition of -glucosidase reduced the intestinal glucose

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digestion as well as absorption, which consequently controlled post-prandial glycaemic response

(plays important role in managing type 2 diabetes) (Balasubramaniam et al., 2013). They also pointed

that flavonols and not flavanols, were the most important contributing factors to the inhibitory action

of lentils against pancreatic lipase and -glucosidase. Myricetin present in black legumes (black

soybean and black turtle bean) showed strong inhibitory activity against -amylase, -glucosidase

and lipase (Tan et al., 2017). Anthocyanins, present especially in beans among legumes are pigments

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that not only impart the color but also have plenty of antioxidant activities responsible for the

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secondary health benefits such as acting as anti-inflammatory, anticarcinogenic agents, prevention of

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cardiovascular disease and having beneficial roles in the diabetes as well as obesity management (He

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and Giusti, 2010).

An epidemiological study indicated that phenolic compounds displayed important function by


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anti-deposition of triglycerides and lowering the levels of free radicals present in the body (Graf et al.,

2005). Oxidation of LDL is generally considered an essential step in atherogenesis (deposition of


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plaques that contain cholesterol as well as lipids in the arterial walls). Alshikh et al. (2015) reported
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the in vitro inhibition of cupric ion induced human low-density lipoprotein (LDL) peroxidation by
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lentils. Germinated pigeon pea exhibits more polyphenolic compounds than ungerminated counterpart

and is a excellent food source for controlling hyperglycemia and lipid peroxidation (Uchegbu and
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Ishiwu, 2016). Hertog et al. (1995) documented the role of phenolic compounds in prevention and
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cure of cancer and heart problems with an epidemiological study. Adebamowo et al. (2005) by their

epidemiological study indicated that among many flavonols and flavonol-rich foods present in the

diet, only the consumption of lentils or beans was associated with lower incidences of breast cancer.

The free phenolic extract of black cowpea seeds at a concentration of 188.1 mg GAE/l have inhibited

proliferation (65%) and reduced viability of mammary (MCF-7) cancer cells in vitro (Gutiérrez-Uribe

et al. 2011). Phenolic compounds in dark beans, especially anthocyanins proved to be the most active

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natural compounds with neuroprotective and antitumoral effects by performing studies on cell

cultures of astrocytes from human glioblastoma, melanoma, renal adenocarcinoma and breast

adenocarcinoma in vitro (Lopez et al., 2013).

Health benefits of phenolic compounds are related to their antioxidant potential and protective

effects on free radical-induced chronic diseases (Xu & Chang, 2010). Antioxidant, antimicrobial and

modulation mechanisms of phenolic compounds have roles in preventing infectious and degenerative

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diseases (Ozcan et al., 2014). Consuming foods rich in phenolic compounds may reduce the risk of

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many health problems. Raw and processed dark beans are good source of natural antioxidants

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beneficial in health problems of high prevalence. Lentil cultivars rich in flavonols can be use as

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functional food and nutraceutical ingredients for promoting health, managing weight and preventing

diabetes (Zhang et al., 2015). Alshikh et al. (2015) reported that bioactivity of all phenolic
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compounds was confirmed by the in vitro inhibition of peroxyl radical induced DNA strand breakage

and therefore lentils should be an integral part of human diet owing to their beneficial health effects.
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García-Lafuente et al. (2014) suggested the role of common bean phenolic extracts having high
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antioxidant as well as high anti-inflammatory activity owing to the reduction in the cytokine mRNA

expression of lipopolysaccharide stimulated macrophages and production of nitrous oxide. These


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results confirmed that common beans should be included in the diet as they are great sources of anti-
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inflammatory agents and function as dietary complement for promotion of human health. High

concentration of anthocyanins (especially delphinidin) and phenolic acids (ferulic acid) reported in
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beans may have health benefits and they are commonly used as functional food ingredients (Mojica et

al., 2015). Beneficial properties of legumes seeds is related to phenolic content and composition

(phenolic acids, flavonoids, tannins) and their role in different bioactivities.

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6. Conclusion and future prospects

The presence of phenolic compounds in legume seeds along with their associated antioxidant

activities make them suitable agents for developing functional foods which can be a part of the daily

diet. Dietary intake of natural antioxidants such as phenolic compounds is an important part of the

defense mechanism in humans. With regard to these activities, along with known nutritional value

(especially as protein sources), it is beneficial to include legumes in significant quantities in the diet

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especially in the Western countries. However, there are still loopholes in the amount of available

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literature and more studies should be encouraged such as those involving clinical studies that could

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establish a strong relationship as well as develop guidelines for the human diet. Another thing that

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needs to be worked at involves the better knowledge of bioavailability of phenolic compounds

present in legumes that is important for the investigation of their health effects (whichever approach
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is employed). Moreover, most of studies done in legumes are based on identifying the aglycones

which are not essential metabolites present in blood owing to their extensive hepatic and intestinal
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conjugation.
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A significant development has been made in the field involving cardiovascular diseases.

Phenolic compounds in sources such as legume seeds can be administered as food supplements as
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their positive roles have been pointed out by several biomarkers that are closely associated with
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cardiovascular disease. However, the evidence for protective effects of phenolic compounds against

neurodegenerative diseases and most cancers is still mainly derived from in vitro studies and animal
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experiments. Still phenolic compounds clearly improve the status of various oxidative stress

biomarkers and improve our health. More clinical trials and epidemiological studies should be carried

for an in depth analysis about the role of phenolic compounds in our health. Moreover, the effect of

processing as well as cooking methods on the content and nature of phenolic compounds in legumes

is an area that requires more investigation. There are many possibilities existing in this area for future

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studies. As legumes are easily available, safe to consume and relatively inexpensive than other foods

academic and governmental institutions can easily invest in studies involving these crops.

Acknowledgments

Authors are thankful to University Grants Commission, New Delhi for providing financial assistance

in the form of Major Research Project.

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Figure 1: Phenolic acids (Hydroxybenzoic acids[a] and hydroxycinnamic acids [b] identified in

legume seeds

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Figure 2: Catechins and procyanidins (a); and anthocyanins (b) identified in legume seeds

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Figure 3: Flavonols (a), flavones (b) and flavanones (c) identified in legume seeds

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Table 1: Total phenolic content and polyphenols reported in various grain legume seeds and their coats
Pulses Total phenolic Technique used Main identified compounds References
content
(i) Dry beans
Commercial bean 19.1 to 48.3 HPLC-DAD p-coumaric acid, ferulic acid and sinapic acid Luthria and Pastor-
varieties mg/100 g DW Corrales (2006)
Bean cultivars 11.2 to 25.3 mg Folin–Denis - Rocha-Guzmán et
CE/g DW al. (2007)

Commercial bean - HPLC-MS Delphinidin, petunidin, malvidin, quercetin Lin et al. (2008)
varieties
Bean coats 146 to 5798 mg Folin- Ciocalteu, Cyanidin-3-glucoside, cyanidin-3-galactoside, Gan et al. (2016)
GAE/100 g DW HPLC malvidin-3-glucoside, peonidin-3-glucoside,
kaempferol, quercetin, rutin, gallic acid

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Common beans 2.4 to 13.5 mol Folin–Ciocalteu, Gallic acid, vanillic acid, protocatechuic acid, Wang et al. (2016)

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trolox/g DW UPLC catechin, epicatechin, p-coumaric acid,
ferulic acid, rutin, quercetin

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Faba bean 117.8 to 157.6 Folin–Ciocalteu - Chaieb et al. (2011)
genotypes mg GAE/g DW
Immature faba 817.0 to 1337.8 Folin–Ciocalteu, Prodelphinidin dimers, (+)-Catechin, (-)- Baginsky et al.
bean seed varieties mg GAE/kg DW HPLC-DAD-MS Epicatechin, Quercetin 3-O-rutinoside, (2013)
Apigenin 7-O-glucoside, Myricetin-3-O-

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glucoside, Quercetin 3-O-glucoside,
Myricetin

Mung bean 26.7 ± 1.4 mg Folin-Ciocalteu, - Zhao et al. (2014)


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Pinto bean GAE/g extract
Black kidney bean 33.4 ± 3.0 mg
Red kidney bean GAE/g extract
32.9 ± 0.1 mg
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GAE/g extract
27.1 ± 3.0 mg
GAE/g extract
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Adzuki bean 90 to 189 mg/g in Folin-Ciocalteu, hydroxycinnamates, procyanidins, gallates, Amarowicz et al


crude extract and HPLC-MS flavonols, dihydroflavonols and (2008)
fractions dihydrochalcones
Kidney beans 0.25 to 35.11 mg Folin–Ciocalteu, pelargonidin, cyanidin, petunidin, delphinidin, Kan et al. (2016)
varieties GAE/g DW UPLC-DAD-MS malvidin
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(ii) Lentils
Lentil varieties - HPLC-MS Protocatechuic acid, catechin, trans-ferulic Dueñas et al. (2006)
(seed coats and acid, gallic acid, procyanidins, epicatechin,
cotyledons) apigenin, myricetin 3- ramnoside, luteolin 7-
CE

glucoside

Lentil varieties 4.9 to 7.8 mg Folin-Ciocalteu - Xu et al. (2007)


GAE/g DW
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Lentil varieties 12 mg GAE/g 4-aminoantipyrine - Han and Baik


DW (2008)
Green lentil 68 mg CE/g DW Folin–Ciocalteu, Catechin glucoside, procyanidin dimer, trans- Amarowicz et al.
Red Lentil 58 mg CE/g crude HPLC-PAD, p-coumaric acid, quercetin diglycoside, trans- (2010) Amarowicz
extract HPLC–ESI-MS ferulic acid, kaempferol glucoside et al (2009)

Green lentil seed 10.3 mg CE/g HCl method - Oomah et al. (2011)
and hull DW and 82.9 mg
CE/g DW
Red lentil seed 12.6 mg CE/g HCl method -
and hull DW and 87.2 mg
CE/g DW

Lentil seed coats - HPLC-MS Gallocatechin, procyanidin C1, malvidin-3- Mirali et al. (2014)
O-galactoside, myricetin-3-O-rhamnoside,
quercetin-3-O-galactoside, kaempferol-3-O-

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glucoside, naringenin, luteolin

Green lentil 5.0 to 7.0 mg Folin–Ciocalteu, Dihydroxybenzoic acid, catechin glucoside,


cultivars GAE/g DW HPLC-MS syringic acid, trans-p-coumaric acid, Zhang et al. (2015)
Red lentil 4.6 to 7.6 mg epicatechin gallate, quercetin-3-glucoside,
cultivars GAE/g DW kaempferol-3-glucoside

Pardina Lentils HPLC-PAC and Dihydroxybenzoic acid, Catechin 3-glucoside, Aguilera et al.
HPLC-MS (ESI) procyanidin, trans-p-coumaric acid, (2010)
kaempferol 3-rutinoside, kaempferol 3-
glucoside and luteolin 3-7-diglucoside

Aguilera et al.
(2011)
Lentil 47.6 ± 5.3 mg Folin-Ciocalteu - Zhao et al (2014)

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GAE/g extract
Lentil cultivars Free: 1.37–5.53 HPLC-DAD-ESI- Catechin, epicatechin, procyanidins B, methyl Alshikh et al.

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mg GAE/g MSn vanillate, procyanidin dimer A, and (2015)
Esterified: 2.32– prodelphinidin dimer A
21.54 mg GAE/g

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Insoluble-
bound:2.55–17.51
mg GAE/g
(iii) Chickpea

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Chickpea variety 0.98 mg GAE/g Folin-Ciocalteu - Xu et al. (2007)
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Chickpea variety 2.2 mg GAE/g 4-aminoantipyrine - Han and Baik
DW (2008)
Chickpea variety 0.54 mg CE/g Folin–Ciocalteu - Fernandez-Orozco
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DW et al. (2009)
Chickpea seed 0.2 to 32.6 mg Folin–Ciocalteu - Segev et al. (2010)
coats and dehulled CE/g DW and 0.4
seeds to 0.8 mg CE/ g
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Chickpea variety HPLC-PAD and Dihydroxybenzoic acid, p-hydroxybenzoic Aguilera et al.


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HPLC-MS acid, trans-p-coumaric acid, pinocembrin, (2011)


quercetin 3-O-rutinoside, genistein hexoside

Chickpea varieties 11.4 to 19.4 mg Folin-Ciocalteu - Nithiyanantham et


TAE/ g DW al. (2012)
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Chickpea cultivars 147 and 183 Folin-Ciocalteu, Gallic acid, chlorogenic acid, catechin, Fratianni et al.
GAE/g DW UPLC quercetin, ferulic acid (2014)

Chickpea 21.9 ± 2.8 mg Folin-Ciocalteu Zhao et al (2014)


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-
GAE/g extract
Chickpea varieties - HPLC-DAD p-Hydroxybenzoic acid, syringic acid, Magalhães et al.
gentisic acid, luteolin-8-C-glucoside, (2017)
myricetin-3-O-rhamnoside, quercetin-3-O-
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galactoside, quercetin-3-O-rhamnoside
(iv) Cowpea
Black seed coated - HPLC-DAD-MS, Delphinidin-3-O-galactoside, delphinidin-3- Ha et al. (2010)
cowpea variety NMR spectroscopy O-glucoside, cyanidin-3-O-galactoside,
cyanidin-3-O-glucoside, petunidin-3-O-
glucoside, pelargonidin-3-O-glucoside,
peonidin-3-O-glucoside, malvidin-3-O-
glucoside

Commercial black 107.3 mg Folin-Ciocalteu, Gallic acid, protocatechuic acid, p- Gutiérrez-Uribe et


coloured cowpeas GAE/100 g DW HPLC–PDA hydroxybenzoic acid, coumaric acid, ferulic al. (2011)
acid

Cowpea varieties - UPLC Proanthocyanidin compounds Ojwang et al.


(2013)

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(v) Pigeon pea
Pigeon pea seeds 786.0 to 1212.0 Folin-Denis - Ferreira et al.
mg GAE/kg DW (2004)
(vi) Peas
Dark peas HPLC-PAD and Protocatechuic , p-hydroxybenzoic, vanillic, Dueñas et al. (2006)
HPLC-MS syringic, trans p-coumaric,cis p-coumaric, and
trans ferulic acid, (-) epigallocatechin,
apigenin 8-C glucoside and quercetin 3-O-
galactoside

Yellow peas 2.5 mg GAE/g 4-aminoantipyrine - Han and Baik


DW (2008)
Pea (coloured 46.36 µg/ g DW HPLC protocatechuic, gentisic, vanillic, caffeic, Troszynska and
seed coat) 7.75 µg/g DW syringic, p-coumaric, ferulic and Ciska (2002)
Pea (white seed p-coumaric

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Grass pea 206 and 213 mg Folin-Ciocalteu, Gallic acid, chlorogenic acid, catechin, Fratianni et al.

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cultivars GAE/g DW UPLC epicatechin, coumaric acid (2014)
Peas, lentils and - HPLC-MS Gallocatechin- (4α→2)-phloroglucinol, Jin et al. (2012)
faba beans gallocatechin, epicatechin-(4β→2)-

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Pea varieties - HPLC-DAD protocatechuic acid, p-Hydroxybenzoic acid Magalhães et al.

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Luteolin-6-C-glucoside, apigenin-8-C- (2017)
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GAE= Gallic acid equivalents, CE= Catechin equivalents, TAE= Tannic acid equivalents
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Table 2: Total flavonoid content (TFC) reported in various grain legume seeds and their coats
Legume seeds/coats Total flavonoid content References
(TFC)
(i) Beans
Black turtle bean (crude extract) 70.2 mg CE/g Tan et al (2017)
Black soybean (crude extract) 49.6 mg CE/g
Kidney beans (26 cultivars) 0.19 and 7.05 mg RE/g Kan et al (2016b)
Red sword bean coat 432 mg CE/ 100g Gan et al (2016)
Pinto bean coat 4477 mg CE/ 100g
Adzuki bean coat 941 mg CE/ 100g
Broad bean coat 689 mg CE/ 100g
Black hyacinth bean coat 694 mg GAE/ 100g
Red Kidney bean coat 2635 mg CE/ 100g
Violet red kidney bean coat 2598 mg CE/ 100g
Big speckled kidney bean coat 3513 mg CE/ 100g

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Small speckled kidney bean coat 3529 mg CE/ 100g
Panda bean coat 546 mg CE/ 100g

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Big runner bean coat 1526 mg CE/ 100g
Small runner bean coat 2093 mg CE/ 100g
Brown string bean 266 mg CE/ 100g

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Climbing bean coat 1019 mg CE/ 100g
Black mung bean coat 288 mg CE/ 100g
Green mung bean coat 330 mg CE/ 100g
Big rice bean coat 1106 mg CE/ 100g

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Small rice bean coat 1524 mg CE/ 100g
Horse gram flour 8.6 mg CE/g Sreerama et al. (2012)
Faba bean (13 genotypes) 5.19 to 8.61 mg RE/g Chaieb et al. (2011)
Black soybean 4.04 mg CE/g Xu et al. (2007)
Yellow soybean cultivars 1.06 to 1.24 mg CE/g
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Black bean cultivars 2.51 to 3.30 mg CE/g
Pinto bean 2.99 mg CE/g
Navy bean 0.92 mg CE/g
Red kidney bean 3.39 mg CE/g
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Small red bean 4.24 mg CE/g


Pink bean 3.65 mg CE/g
(ii) Lentils
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Red lentil cultivars 0.60 to 1.62 mg CE/g Zhang et al (2015)


Green lentil cultivars 0.66 to 1.98 mg CE/g
Lentil cultivars:
Free phenolic compounds 0.01–0.80 mg CE/g Alshikh et al. (2015)
Esterified phenolic compounds 0.36–4.13 mg CE/g
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Insoluble-bound phenolic compounds 0.08-2.95 mg CE/g


Lentil (11 cultivars) 2840 to 6870 μg/g Xu and Chang (2010)
Lentils cultivars 3.04 to 4.54 mg CE/g Xu et al. (2007)
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(iii) Pea
Pea seed coat (Mottled) 104 mg CE/ 100g Gan et al (2016)
Pea 7.93 mg RE/g Nithiyanantham et al.
(2012)
Yellow pea 0.09 to 0.17 mg CE/g Xu et al. (2007)
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Green pea 0.05 to 0.15 mg CE/g


(iv) Cowpea
Red cowpea coat 1383 mg CE/ 100g Gan et al (2016)
Mottled cowpea coat 2303 mg CE/ 100g
Cowpea flour 7.24 mg CE/g Sreerama et al. (2012)
Black cowpeas (whole seed) 173.17 mg QE/ 100g Gutiérrez-Uribe et al.
Black cowpeas (Seed coat) 1730 mg QE/ 100g (2011)
Black cowpeas (Cotyledons) 0.09 mg QE/ 100g
(vi) Chickpea
Chickpea 10.65 mg RE/g Nithiyanantham et al.
(2012)
Chickpea flour 4.8 mg CE/g Sreerama et al. (2012)
Chickpea whole seeds (17 lines) 0.1 to 1.08 mg CE/g DW Segev et al. (2010)
Chickpea seed coats (17 lines) 0.1 to 7.2 mg CE/g DW
Chickpea dehulled seeds (17 lines) 0.08 to 0.11 mg CE/ g DW
Chickpea 0.72 mg CE/g Xu et al. (2007)

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(vii) Pigeon pea
Pigeon pea seed coat 2651 mg CE/ 100g Gan et al (2016)
(v) Peanuts
Seed coat (6 peanut varieties) 31.68 - 85.17 mg CE/g Attree et al (2015)
Raw kernel (6 peanut varieties ) 0.97 - 2.84 mg CE/g
Cotyledons 0.18 – 0.30 mg CE/g

RE= rutin equivalents, QE=quercetin equivalents, CE= Catechin equivalents

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Table 3: Condensed tannins content reported in various grain legume seeds and their coats

Legume seeds/coats Condensed tannins References


content (CTC)
(i) Beans
Broad bean (crude extract) 5.96 A500/g Amarowicz and Shahidi
(2017)
Black turtle bean (crude extract) 40.6 mg CE/g Tan et al (2017)
Black soybean (crude extract) 20.6 mg CE/g
Kidney beans coats (26 cultivars) 10.52 to 57.13 mg PAE/g Kan et al (2016a)
Adzuki bean coat 13.8 mg CE/ g Gan et al (2016)
Broad bean coat 18.0 mg CE/ g
Red Kidney bean coat 46.7 mg CE/ g

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Violet red kidney bean coat 57.9 mg CE/ g
Big speckled kidney bean coat 79.5 mg CE/ g

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Small speckled kidney bean coat 79.9 mg CE/ g
Panda bean coat 12.5 mg CE/ g
Big runner bean coat 28.1 mg CE/ g

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Small runner bean coat 44.7 mg CE/ g
Brown string bean 5.26 mg CE/ g
Climbing bean coat 19.0 mg CE/ g
Big rice bean coat 22.5 mg CE/ g

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Small rice bean coat 26.7 mg CE/ g
Bean seed coats (15 cultivars) 60.6 to 369.3 mg GAE/g Mojica et al (2015)
coat
Raw cranberry bean (regular darkening) 1.63 – 2.89 mg PAC/g Chen et al (2015b)
Raw cranberry bean (non-darkening ) 0.02 – 0.05 mg PAC/g
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Fava beans (10 varieties) 309.28 to 958.77 mg CE/kg Baginsky et al. (2013)
Faba bean 654 mg/100 g Jin et al (2012)
Horse gram flour 3.8 mg CE/g Sreerama et al. (2012)
Black bean cultivars 4.09 - 5.73 mg CE/g Xu et al. (2007)
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Yellow soybean cultivars 0.37 - 0.79 mg CE/g


Pinto bean 3.23 mg CE/g
Navy bean 0.47 mg CE/g
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Red kidney bean 2.87 mg CE/g


Small red bean 5.16 mg CE/g
Pink bean 3.55 mg CE/g
Black soybean 1.96 mg CE/g
Adzuki bean 0.483 A500/mg Amarowicz et al (2004)
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Red bean 0.434 A500/mg


Faba bean 0.416 A500/mg
Broad bean 0.093 A500/mg
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(ii) Lentils
Red lentil cultivars 3.00 to 5.82 mg CE/g Zhang et al (2015)
Green lentil cultivars 3.36 to 7.80 mg CE/g
Lentil cultivars:
Free phenolic compounds 0.40–2.67 mg CE/g Alshikh et al. (2015)
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Esterified phenolic compounds 0.64–10.5 mg CE/g


Insoluble-bound phenolic compounds 0.03–4.28 mg CE/g
Lentil (3 cultivars) 269 - 378 mg/100 g Jin et al 2012
Green lentil 93 A500/g Amarowicz et al. (2010)
Red lentil 70 A500/g Amarowicz et al. (2009)
Lentils cultivars 3.73 to 10.20 mg CE/g Xu et al. (2007)
Red lentil 0.577 A500/mg Amarowicz et al (2004)

(iii) Pea
Pea seed coat (Mottled) 7.56 mg CE/ g Gan et al (2016)
Pea (two cultivars) 294 & 367 mg/100 g Jin et al (2012)
Pea (acetone extract) 1.03 mg TAE /g Nithiyanantham et al.
Pea (methanol extract) 6.85 mg TAE /g (2012)
Yellow pea 0.22 to 0.59 mg CE/g Xu et al. (2007)
Green pea 0.23 to 0.61 mg CE/g
(iv) Cowpea

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Red cowpea coat 41.7 mg CE/ g Gan et al (2016)
Mottled cowpea coat 15.8 mg CE/ g
Cowpea varieties 2.2 – 6.3 mg/g Ojwang et al. (2013)
Cowpea flour 2.13 mg CE/g Sreerama et al. (2012)
(v) Chickpea
Chickpea 0.52 mg CE/g Xu et al. (2007)
Chickpea (acetone extract) 9.48 mg TAE /g Nithiyanantham et al.
Chickpea (methanol extract) 10.84 mg TAE /g (2012)
Chickpea flour 5.12 mg CE/g Sreerama et al. (2012)
(vi) Pigeon pea
Pigeon pea seed coat 71.8 mg CE/ g Gan et al (2016)
Pigeon pea (3 accessions) 348 to 641 mg /kg Ferreira et al. (2004)
(vii) Peanuts
Seed coat (6 peanut varieties) 29.7 - 84.7 mg CE/g Attree et al (2015)
Raw kernel (6 peanut varieties ) 2.88 - 4.73 mg CE/g
1.74 – 3.75 mg CE/g

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proanthocyanidin content

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Table 4: Antioxidant activity of various grain legume seeds and their coats
Seeds/seed coats Antioxidant activity References
(i) Dry beans
Broad bean (crude extract) 0.583 mmol TE/g Amarowicz and Shahidi (2017)
Black turtle bean (crude extract) 49.2 μmol TE/g Tan et al (2017)
Black soybean (crude extract) 23.7 μmol TE/g
Black bean 5.9 μmol Trolox/g Wang et al. (2016)
Kidney bean 5.2 μmol Trolox/g
Mung bean 3.7 μmol Trolox/g
Red bean 4.9 μmol Trolox/g
Red kidney bean 5.5 μmol Trolox/g

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Soy bean 3.2 μmol Trolox/g
Red sword bean coat 699 μmol TE/g Gan et al (2016)

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Pinto bean coat 237 μmol TE/g
Adzuki bean coat 95.3 μmol TE/g
Broad bean coat 113 μmol TE/g

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Black hyacinth bean coat 55.5 μmol TE/g
Red Kidney bean coat 308 μmol TE/g
Violet red kidney bean coat 212 μmol TE/g
Big speckled kidney bean coat 288 μmol TE/g
282 μmol TE/g

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Small speckled kidney bean coat
Panda bean coat 71.1 μmol TE/g
Big runner bean coat 118 μmol TE/g
Small runner bean coat 203 μmol TE/g
Brown string bean 37 μmol TE/g
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Climbing bean coat 94.7 μmol TE/g
Black mung bean coat 29.0 μmol TE/g
Green mung bean coat 29.7 μmol TE/g
Big rice bean coat 95.2 μmol TE/g
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Small rice bean coat 140 μmol TE/g


Kidney beans coats (26 cultivars) 4.00 to 491.33 μmol Trolox/g Kan et al (2016a)
Kidney beans (26 cultivars) 1.07 to 7.48 μmol Trolox/g Kan et al (2016b)
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Bean seed coats (15 cultivars) 185.2 to 233.9 mmol TE/g Mojica et al (2015)
Cranberry bean seed coat (dark coloured) 125.28-287.87 μmol TE/g Chen et al (2015a)
Cranberry bean seed coat (light coloured) 2.11- 10.09 μmol TE/g
Mung bean 304 U/g Zhao et al. (2014)
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Pinto bean 567 U/g


Black kidney bean 602 U/g
Red kidney bean 516 U/g
Small red bean 622 U/g
217.97 μmol TE/g
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White kidney bean García-Lafuente et al (2014)


Round purple bean 304.24 μmol TE/g
Horse gram flour IC50 22.9 μg/ml Sreerama et al. (2012)
Raw Cannellini bean flour 38.3 μmol Trolox/g Aguilera et al. (2011)
Pinto bean flour 96.2 μmol Trolox/g
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Faba bean (13 genotypes, whole seeds) 2.15 -28.60 % Chaieb et al. (2011)
Adzuki bean (crude extract) 1.76 μmol Trolox/mg Amarowicz et al (2008)
Soybean 10.2 μmol TE/g Han and Baik (2008)
Common beans varieties 13.30 to 92.73 μmol of TE/g Xu et al. (2007)
Black soybean 131.3 μmol TE/g
Yellow soybean cultivars 35.10 to 44.23 μmol TE/g
Adzuki bean 1.76 μmol Trolox/mg Amarowicz et al (2004)
Red bean 1.49 μmol Trolox/mg
Faba bean 0.88 μmol Trolox/mg
Broad bean 0.58 μmol Trolox/mg
(ii) Lentils
Lentil cultivars:
Free phenolic compounds 0.01–0.48 μmol TE/g Alshikh et al. (2015)
Esterified phenolic compounds 0.07–0.44 μmol TE/g
Insoluble-bound phenolic compounds 0.04–0.42 μmol TE/g
Two lentil cultivars 1.70 & 2.22 mg EC 50 Fratianni et al. (2014)

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Lentil 721 U/g Zhao et al (2014)
Green lentil seed 136.1 μmol TE/g Oomah et al. (2011)
Green lentil hull 955.6 μmol TE/g
Red lentil seed 134.8 μmol TE/g
Red lentil hull 1040.5 μmol TE/g
Raw Pardina Lentils 66.97 μmol Trolox/g Aguilera et al. (2010)
Lentil (11 cultivars) 38.9 to 82.0 μmol TE/g Xu and Chang (2010)
Green lentil (crude extract) 0.75 μmol Trolox eq./mg Amarowicz et al. (2010)
Red lentil (crude extract) 0.68 μmol Trolox/mg Amarowicz et al. (2009)
Pardina Lentils 14.8 μmol TE/g Han and Baik (2008)
Crimson lentils 14.0 μmol TE/g
Lentils cultivars 59.55 to 95.19 μmol TE/g Xu et al. (2007)
Lentil seed coat 0.05- 0.07 mg EC 50 Dueñas et al. (2006)
Lentil cotyledons 21- 29 mg EC 50
Red lentil 0.68 μmol Trolox/mg Amarowicz et al (2004)

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(iii) Pea
Pea seed coat (Mottled) 35.5 μmol TE/g Gan et al (2016)

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Two grass pea cultivars 18.53 & 24.55 mg EC 50 Fratianni et al. (2014)
Pea (acetone extract) 29.40 g /g DPPH Nithiyanantham et al. (2012)
Pea (methanol extract) 13.67 g /g DPPH

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Yellow pea 10.36 μmol TE/g Oomah et al. (2011)
Yellow pea hull 22.89 μmol TE/g
Yellow peas 3.4 μmol TE/g Han and Baik (2008)
Green peas 1.8 μmol TE/g
Yellow pea 3.26 to 12.80 μmol TE/g Xu et al. (2007)

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Green pea 1.73 to 9.95 μmol TE/g
Pea 0.30 μmol Trolox/mg Amarowicz et al (2004)
(iv) Cowpea
Red cowpea coat 112 μmol TE/g Gan et al (2016)
AN
Mottled cowpea coat 211 μmol TE/g
Cowpea flour IC50 48.7 μg/ml Sreerama et al. (2012)
Black cowpeas (whole seed) 13.49 μmol TE/g Gutiérrez-Uribe et al. (2011)
Black cowpeas (Seed coat) 15.47 μmol TE/g
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Black cowpeas (Cotyledons) 13.94 μmol TE/g


(v) Chickpea
Chickpea 3.4 μmol Trolox/g Wang et al. (2016)
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Two Chickpea cultivars 47.90 and 77.09 mg EC 50 Fratianni et al. (2014)


Chickpea (acetone extract) 3.72 g /g DPPH Nithiyanantham et al. (2012)
Chickpea (methanol extract) 12.17 g /g DPPH
Chickpea flour IC50 31.4 μg/ml Sreerama et al. (2012)
Chickpea whole seeds (17 lines) 0.08 to 2.59 mmol TE/ 100g Segev et al. (2010)
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Chickpea seed coats (17 lines) 0.06 to 14.7 mmol TE/ 100g
Chickpea dehulled seeds (17 lines) 0.06 to 0.19 mmol TE/ 100g
Raw chickpea seeds 26.39 μmol Trolox/g Fernandez-Orozco et al. (2009)
Germinated chickpeas 29.66 μmol Trolox/g
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Fermented chicpeas 34.10 μmol Trolox/g


Chickpea 2.0 μmol TE/g Han and Baik (2008)
Chickpea 9.26 μmol TE/g Xu et al. (2007)
(vi) Pigeon pea
AC

Pigeon pea 52.1% Uchegbu and Ishiwu (2016)


Pigeon pea (germinated) 85.2%
Pigeon pea seed coat 206 μmol TE/g Gan et al (2016)
(vii) Peanuts
Seed coat (6 peanut varieties) 25.8 - 28.8 μmol TE/g Attree et al (2015)
Raw kernel (6 peanut varieties ) 10.8 - 28.9 μmol TE/g
Cotyledons 1.5 – 2.72 μmol TE/g
U= Unit of total antioxidant capacity (the amount of antioxidants required to make absorbance increase 0.01 in 1 mL of reaction liquid
at 37 C), TE=Trolox equivalents.

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Graphical abstract
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Highlights

 Phenolics are commonly identified in seeds of edible legumes

 These compounds include phenolic acids, flavonoids and tannins

 Distributed differently in seed coat and cotyledon of legumes

 Phenolics exert a wide range of health benefits

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