International Journal of Food Sciences and Nutrition

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Effect of sprouting on nutritional quality of pulses

Daniela Erba, Donato Angelino, Alessandra Marti, Federica Manini, Franco

Faoro, Federico Morreale, Nicoletta Pellegrini & Maria Cristina Casiraghi

To cite this article: Daniela Erba, Donato Angelino, Alessandra Marti, Federica Manini, Franco
Faoro, Federico Morreale, Nicoletta Pellegrini & Maria Cristina Casiraghi (2018): Effect of
sprouting on nutritional quality of pulses, International Journal of Food Sciences and Nutrition, DOI:
10.1080/09637486.2018.1478393

To link to this article: https://doi.org/10.1080/09637486.2018.1478393

Published online: 31 May 2018.

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INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION
https://doi.org/10.1080/09637486.2018.1478393

RESEARCH ARTICLE

Effect of sprouting on nutritional quality of pulses

Daniela Erbaa, Donato Angelinob , Alessandra Martia, Federica Maninia, Franco Faoroc,
Federico Morrealeb, Nicoletta Pellegrinib and Maria Cristina Casiraghia
a
Department of Food, Environmental and Nutritional Sciences (DeFENS), University of Milan, Milano, Italy; bHuman Nutrition Unit,
Department of Food and Drug, University of Parma, Parco Area delle Scienze, Parma, Italy; cDepartment of Agricultural and
Environmental Sciences - Production, Landscape, Agroenergy, University of Milan, Milano, Italy

ABSTRACT ARTICLE HISTORY

In order to investigate the nutritional quality of industrial-scale sprouted versus unsprouted Received 30 January 2018
chickpeas and green peas, before and after cooking, the ultrastructure, chemical composition, Revised 14 May 2018
antioxidant capacity, starch digestibility, mineral content and accessibility were analysed. Accepted 16 May 2018
Sprouting did not deeply affect raw seed structure, although after cooking starch granules
KEYWORDS
appeared more porous and swelled. Chemical composition of raw sprouted seeds was not Sprouting; pulses; mineral
strongly affected, excepting an increase in protein (both pulses), and in free sugars (in peas; accessibility; starch
þ10% and þ80%, respectively, p < .05). The industrial sprouting favoured phytic acid leaching in digestibility; antioxidant
cooking water (35% in seeds, compared to unsprouted cooked ones, p < .05), and promoted capacity; nutritional
antioxidant capacity reductions in raw and cooked seeds (10% and 37%, respectively, p < .05). composition
In conclusion, sprouting on an industrial-scale induced mild structural modifications in chickpeas
and peas, sufficient to reduce antinutritional factors, without strongly influencing their nutritional
quality. These products could represent nutritionally interesting ingredients for different dietary
patterns as well as for enriched cereal-based foods.

Introduction to pulses (Patterson et al. 2017). Among those

technological processes, sprouting (or germination) is
Pulses have been long known for their nutritional and
very popular in the marketplace and represents a
health-promoting properties, being a good source of
fibre, proteins, antioxidant compounds – including re-emerging trend in healthy foods, thanks to the
phenolic acids, polyphenols and flavonoids – and positive effects on the enhancement of the nutritional
having a low-glycaemic index (Hall et al. 2017). properties (Ghavidel and Prakash 2007) and taste
Therefore, pulses can be consumed as a whole or used (Roland et al. 2017).
as ingredient in many food preparations intended for Traditionally, germination is a process performed at
the general population or particularly for special diets the household level. The basic process consists of
such as vegetarian, vegan or gluten-free. Despite that, steeping grains in water until they reach the moisture
the consumption of pulses is still underexploited by content needed to initiate the seedling. After the
the Western consumers, due to the presence of anti- steeping water is drained, the seeds germinate. Some
nutrients, such as phytic acid, trypsin inhibitors and challenges – including the safety risk and the process
some undigested oligosaccharides, which are respon- reproducibility – need to be overcome to perform the
sible, for example, for digestive discomfort (Hall et al. process at an industrial scale and deliver a safe
2017). Finally, when included in food preparation, the product with consistent features. At this regard, a
presence of off-flavours may discourage the consump- germination plant has been developed by Buhler AG
tion of pulses (Roland et al. 2017). Considering these (Uzwil, Switzerland), in which grains are partially ger-
aspects and following the scientific evidence to recom- minated under controlled conditions (i.e. temperature
mend an increased consumption of pulses to improve and relative humidity) and stabilised through drying
health, several processes – including soaking, dehul- with hot air to extend the product shelf-life. The
ling, extrusion and fermentation – have been applied control of the process seems the only way to balance

CONTACT Daniela Erba daniela.erba@unimi.it Department of Food, Environmental and Nutritional Sciences (DeFENS), University of Milan, Via
Celoria 2, 20133 Milano, Italy
Supplemental data for this article can be accessed here.
ß 2018 Informa UK Limited, trading as Taylor & Francis Group
2 D. ERBA ET AL.

the nutritional and sensory improvements with the evaluated on porridge prepared by cooking in boiling
maintenance of flour performance and to ensure con- water (200 mL) for 3 min ground pulses (40 g).
sistent functionality to the product (Marti et al. 2018). Intact seeds were also processed to simulate domes-
Recently, partial germination has been carried out on tic preparation. Briefly, pulses were soaked in excess of
chickpeas, and the resulting flour has been proposed plain tap water, for 12 h at room temperature (hence-
as an interesting ingredient to produce cereal-based forth “soaked”). After draining, samples were cooked in
foods with improved nutritional and rheological plain tap boiling water for 45 min in 1:3 w/w propor-
characteristics (Marengo et al. 2017). However, the tion (henceforth “cooked”). Soaking and cooking treat-
nutritional qualities of partially sprouted pulses remain ments were replicated twice, and final samples
scarcely addressed so far. combined. Both soaked and cooked samples were used
The present study investigates the impact of sprout- as such for light microscopy or homogenised (7011S-
ing – carried out at an industrial-scale level and under Waring Blender Commercial, Torrington, CT, USA) for
controlled conditions – on the nutritional profile of further analyses.
chickpeas and green peas. To this aim, chemical
composition, microscopy features, antioxidant Light microscopy
capacity, starch digestibility, total content (Ca, Mg, Fe,
Zn and P) and accessibility of minerals (Ca and Mg) Soaked and cooked intact seeds were sectioned with a
were assessed in both unsprouted and sprouted pulses, vibratome in 20–30 mm thick sections. These were
also taking into consideration the effect of cooking on stained with 0.1% water solution of toluidine blue for
nutritional features. 1 min, or Lugol’s iodine for 5 min. Then, the samples
were observed by an Olympus BX50 light microscope
(Olympus, Tokyo, Japan) equipped with Nomarski
Experimental differential interference contrast.
Chemicals and reagents
Cellulose (powder from spruce), ABTS [2,2-azino-bis(3- Chemical composition
ethylbenzothiazoline-6-sulphonic acid) diammonium Determination of chemical composition was performed
salt], Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2- on the unsprouted and sprouted raw pulses. Moisture,
carboxylic acid), potassium persulphate (dipotassium starch, protein, lipid and ash contents were assessed by
peroxydisulphate), ethanol at analytical grade, enzymes, AACC standard methods (44-15.02, 76-13.01, 46-12.01,
bile salts and phytic acid were purchased from Sigma 30-10.01 and 08-01.01, respectively; AACC 2001).
Chemical Co. (St. Louis, MO, USA) and chemicals from Sugars were evaluated by HPLC Anion Exchange
Merck KGaA (Darmstadt, Germany). Milli Q water was Chromatography with Pulsed Amperometric Detection
used throughout the experiments. (HPAEC-PAD; Englyst et al. 2000). Total, soluble and
insoluble dietary fibre content was evaluated by the
Materials enzymatic–gravimetric procedure (AOAC International
1995). The analysis of phytic acid was performed by
Chickpea and green peas were provided by Molino HPLC with spectrophotometric detection as previously
Quaglia (Vighizzolo d’Este, Italy). Samples were described on raw and cooked pulses (Erba et al. 2017).
soaked in water in 1:2 proportion (w/v) for 24 h and Analyses of moisture, starch, protein, lipid, ash and
then germinated – in darkness for 3 days, at 22  C phytic acid contents were repeated four times and data
and 90% relative humidity – in an industrial sprouting expressed as g kg1 on the dry weight (d.w.).
plant (B€uhler AG, Uzwil, Switzerland). After sprout-
ing, grains were dried at 50  C for 10 h until reaching
a final moisture of about 8%, and stored at room Total mineral content and Ca and Mg
temperature. accessibilities
Experimental plan is summarised in Supplementary Total contents of Ca, Mg, Fe, Zn and P were deter-
Figure S1. An aliquot of unsprouted and sprouted mined on raw and cooked samples (both sprouted
pulses was ground into powder (henceforth “raw”; < 0. and unsprouted pulses) and, after an enzymatic diges-
5 mm particle size) in a pin mill (B€ uhler AG, Uzwil, tion of cooked samples, on the supernatant solutions
Switzerland) and used as such for the evaluation of containing the soluble mineral fraction. To investigate
chemical composition, phytic acid, mineral content the uptake of minerals from tap water during the
and antioxidant capacity. Starch digestibility was domestic treatment, mineral levels of tap water were
 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION 3

measured by atomic absorption spectroscopy. For total Statistical analysis

mineral analysis, samples were dry-ashed (550  C,
Analysis of variance (ANOVA) was performed utilis-
overnight) and dissolved with an acid solution, as ing Statgraphics XV version 15.1.02 (StatPoint Inc.,
previously reported (Erba et al. 2011). For Ca and Mg Warrenton, VA, USA) to assess the effects of sprout-
accessibility, cooked (both sprouted and unsprouted) ing and processing; differences between means were
samples were subjected to two enzymatic digestion further evaluated by Tukey’s post hoc test (p < .05).
stages, i.e. gastric and intestinal, to simulate the The effect of sprouting on the accessibility of Ca and
human gastro-intestinal system. Minerals in the Mg in cooked pulses and on the total antioxidant
soluble fractions were analysed (Erba et al. 2017). capacity was analysed by Student’s t-test (p < .05).
Mineral concentrations were determined by Atomic
Absorption Spectroscopy (AAnalyst800 Perkin Elmer, Results and discussion
Waltham, MA, USA), against standard curves (Ca:
0.5–4 mg L1; Mg: 0.1–0.4 mg L1; Fe: 1–4 mg L1; Microscopy features
Zn: 0.1–0.8 mg L1), using lanthanum solution to Figure 1 shows the effect of sprouting on the micro-
avoid interferences for Ca and Mg analyses (1 g kg1 structure of the whole pulses – soaked and cooked –
w/v). Phosphorous (P) was determined by a colorimet- captured under polarised light conditions. In order to
ric method (Erba et al. 2011). The accuracy of mineral highlight the cell walls, samples were stained with a
analyses was checked by using certified values of refer- solution of Toluidine blue, which is a generic dye for
ence material (NCS ZC 85006). All analyses were plant tissues.
performed in triplicate, and results were expressed as The arrangement of the cells in the seed tissue was
total mineral content (mg kg1 d.w., mean ± SD). clearly visible in all the samples, regardless of the applied
Accessible Ca and Mg were expressed as percentage of process. The sprouting conditions adopted in the present
soluble Ca and Mg with respect to total content. study did not dramatically affect the seed structure of
either pea (Figure 1(b)) or chickpea (Figure 1(f)), when
compared with the unsprouted pulses [Figure 1(a,e) for
In vitro starch digestibility peas and chickpeas, respectively]. The cell walls were vis-
The in vitro starch digestibility was assessed on ible even after cooking in both unsprouted [Figure 1(c)
cooked pulses (as seeds or porridge), according to for peas and Figure 1(g) for chickpeas] and sprouted
Englyst’s method (Englyst et al. 2000). Rapidly (RDS) pulses [Figure 1(d) for peas and Figure 1(h) for chick-
and slowly (SDS) digestible as well as total starch frac- peas], appearing as grey lines. This observation suggests
that they had and maintained a high degree of ordered
tions were calculated from the glucose released data
structure (Brummer et al. 2015). Interestingly, in sprout-
(at 20 and 120 min of hydrolysis, respectively), deter-
ing seeds, cells appeared slightly more stained by
mined by HPLC. Six set of data from three independ-
Toluidine, indicating the presence of mobilised proteins
ent cooking trials were averaged. RDS and SDS
(more accessible to the stain).
fractions were expressed as the percentage of available
The majority of the interior of the cells was occupied
starch (AVST ¼ RDS þ SDS).
by starch granules, which in Figure 2 appear in dark as
samples were stained with Lugol. The surface of the gran-
Determination of total antioxidant capacity ules was generally smooth without any fissures, cracks or
pores. A granule size between 2.72 mm and 31.8 mm was
The total antioxidant capacity (TAC) of raw, soaked observed for pea starches (Aggarwal et al. 2004). The
and cooked samples was determined with a direct large oval to small spherical-shaped granules were
measurement according to Açar et al. (2009) without reported to be present in starch from different chickpea
any sample preparation. If required, samples were cultivars (Brummer et al. 2015).
diluted with cellulose powder, which was found to Starch granules of sprouted pulses (Figure
be inert towards ABTS reagent (Serpen et al. 2007). 2(b,d,f,h)) appeared darker compared to the related
The total antioxidant capacity was expressed as unsprouted samples (Figure 2(a,c,e,g)) suggesting the
mmol of Trolox equivalent antioxidant capacity formation of a more porous structure after sprouting.
(TEAC) per kg of dry sample, by means of an at Cooking of whole pulses in boiling water resulted in
least five points dose-response curve. The analyses the swelling of starch granules, but not in the disrup-
were repeated six times, and data were presented tion of the cell walls or the starch granules,
as mean ± SD. consistently with other studies (Brummer et al. 2015).
4 D. ERBA ET AL.

Figure 1. Storage seed tissues, stained by Toluidine blue, in unsprouted (a, c, e, g) and sprouted (b, d, f, h) peas (upper panels)
and chickpeas (lower panels), after soaking (left panels) and cooking (right panels); in all samples, cells appeared limited by a well
preserved cell wall and filled with starch granules, which were particularly swollen in cooked seeds (arrows); all bars ¼ 100 mm.

Figure 2. Appearance of starch granules after Lugol staining in unsprouted (a, c, e, g) and sprouted (b, d, f, h) peas (upper panels)
and chickpeas (lower panels), after soaking (left panels, isolated granules) and cooking (right panels, arrows point at granules inside
storage cells); granules from unsprouted seeds were not penetrated by the stain and appeared whitish; all bars ¼ 50 mm.

Chemical composition
Other studies reported an increase in protein in germi-
The partial germination process did not promote signifi- nated grains, which has been attributed to the synthesis
cant differences in pulses in terms of starch, lipid and of enzymes (for example, proteases) during germination
fibre content, except for a protein increase (Table 1). (Masood et al. 2014) or to the compositional changes
 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION 5

Table 1. Effect of sprouting on the chemical composition of treatments for removing a-galactosides, oligosacchar-
raw chickpea and green peaa. ides derived from sucrose, containing 1–3 units of
Chickpea Green pea galactose linked by a-1,6 linkages. These oligosacchar-
Unsprouted Sprouted Unsprouted Sprouted ides, commonly present in legumes and rapidly fer-
Protein 186 ± 3d 202 ± 1b 194 ± 0c 216 ± 2a mented by the human colonic microbiota, are
Lipid 73 ± 2a 73 ± 5a 24 ± 1b 25 ± 2b
Starch 518 ± 44a 545 ± 78a 590 ± 63a 539 ± 70a responsible for flatulence in individuals that do not
Free sugars 56 ± 1b 54 ± 3b 46 ± 7b 83 ± 3a consume pulses on a regular basis (Hall et al. 2017).
Ash 28 ± 1a 26 ± 0b 27 ± 0ab 27 ± 0ab
Soluble fibre 12 ± 2c 19 ± 3bc 31 ± 5a 27 ± 2ab The increase in the relative percentage of sucrose in
Insoluble fibre 169 ± 3a 172 ± 1a 155 ± 11a 160 ± 8a the sprouted products was likely the result of the com-
a
Values are presented as mean ± SD (n ¼ 4) and expressed as g kg1 dry positional changes following the degradation of raffin-
weight. Data in the same row with different letters are significantly
different (p < .05). ose, probably attributable to the endogenous or
microbial enzymatic activities developed during
sprouting (Cai et al. 1997; M€akinen and Arendt 2015).
Chickpea Sprouted Chickpea
2% The sucrose increment in flour from germinated
2% 2%
chickpeas improved the leavening properties of dough
enriched with sprouted pulses (Marengo et al. 2017)
40%
and may positively contribute to the sensory charac-
teristics of derived products.
56%
98% glucose Germination is reported as the more effective pro-
fructose cess for reducing phytic acid in pulses (Patterson et al.
sucrose 2017). This compound is considered an antinutritional
Green Pea Sprouted Green Pea
raffinose
factor with a marked chelating ability – in particular
2% 3% 7% 1% for calcium – and it is linked to the inhibition of
maltose
digestive enzymes, such as proteases and a-amylases.
In this study, myo-inositol hexaphosphate (IP6) was
35% 30% the only inositol phosphate found. As observed in pre-
vious studies (Egli et al. 2002), after germination the
30% 92%
content of phytic acid in chickpeas significantly
decreased by 5% (19 ± 0 vs. 18 ± 0 g kg1 d.w.,
Figure 3. Effect of sprouting on the percentage of free sugars p < .05; Table 2). Such an effect was probably related
in raw pulses. to the activation of endogenous phytase (Egli et al.
2002), occurring during the initial period of
germination.
following the degradation of other constituents
(Bau et al. 1997).
The fibre content did not change between Table 2. Total minerala and phytic acidb contents in raw and
cooked chickpea and green pea (unsprouted and sprouted).
unsprouted and sprouted pulse samples. This disagrees
Unsprouted Sprouted
with studies showing that the germination process has
a significant impact on dietary fibre fractions in pulses Raw Cooked Raw Cooked
Chickpea
(Martın-Cabrejas et al. 2003; Masood et al. 2014; Ca 791 ± 17c 2434 ± 105a 545 ± 72d 2052 ± 57b
Duenas et al. 2016). These differences might be Mg 942 ± 77a 929 ± 15a 978 ± 54a 705 ± 2b
Fe 70 ± 2a 38 ± 2c 63 ± 2b 75 ± 4a
explained by considering the different type of legume Zn 25 ± 1c 53 ± 1a 27 ± 1c 39 ± 1b
(chickpeas, beans, etc.) and the germination conditions P 4037 ± 28a 3982 ± 29a 4160 ± 224a 2726 ± 37b
(e.g. sprouting time, temperature and relative humid- Phytic acid 19 ± 0a 18 ± 0b 18 ± 0b 11 ± 0c
Green pea
ity, light/darkness conditions, drying temperature; Ca 769 ± 56b 1967 ± 86a 824 ± 127b 1966 ± 52a
Martın-Cabrejas et al. 2003). Mg 958 ± 27a 855 ± 29b 870 ± 26b 637 ± 21c
Fe 62 ± 3a 40 ± 5b 58 ± 1a 42 ± 9b
In chickpeas, the total amount of free sugars did Zn 38 ± 1a 32 ± 1b 38 ± 2a 36 ± 0a
not change, whereas a significant increase (p < .05) P 4172 ± 167a 3928 ± 159a 4560 ± 73a 2592 ± 77b
Phytic acid 17 ± 0a 16 ± 0a 17 ± 1a 11 ± 0b
was found in peas. As regards the qualitative distribu-
Data in the same row with different letters are significantly different
tion of sugars (Figure 3), we evidenced an increase in (p < .05).
sucrose and the disappearance of raffinose after the a
Values are presented as mean ± SD (n ¼ 3) and expressed as mg kg1
dry weight.
sprouting of both pulses. Concerning to raffinose, b
Values are presented as mean ± SD (n ¼ 4) and expressed as g kg1
soaking and sprouting are the most efficient biological dry weight.
6 D. ERBA ET AL.

In both germinated and cooked seeds, a 35% After cooking, P was significantly reduced in both
reduction (p < .01) in IP6 levels was evidenced. The sprouted pulses, in accordance with the decrease of
decrease in IP6 suggests a potential leaching of the phytic acid (Table 2), suggesting that the release of P
phytate into the soaking and/or cooking water, owing into the cooking water is favoured by seed structure
to its water solubility. Compared to unsprouted pulses, modification.
the leaching effect was more pronounced in the The total mineral contents of raw unsprouted
sprouted ones, likely due to their porous structure pulses were consistent with data reported in the litera-
(Figures 1(b,d,f,h)). ture (Ray et al. 2014), considering the known variabil-
ity due to different accessions of legumes and the
influences of the agronomic practices (Dalfollo Ribeiro
Total mineral contents and Ca and et al. 2012).
Mg accessibility In raw pulses, mineral contents were only partially
Total mineral content was influenced by both the type affected by germination; Ca and Fe significantly
of pulses (chickpeas or green peas) and the treatments decreased in chickpeas and Mg in green peas.
(sprouting and cooking; Table 2). Cooking led to a Contrasting results have been reported on the effect of
more than 2-fold increase in Ca levels in both legumes, sprouting on total mineral contents in raw pulses.
independently by the sprouting process (p < .05). This Some studies showed significant mineral decreases
increase was likely due to the Ca-uptake from the tap after sprouting, likely due to the leaching of solid mat-
water (whose Ca level was 72.5 mgL1) and a consist- ter in soaking water (Ghavidel and Prakash 2007;
ent increase of legume weight of about 2.5-fold. Tap Audu and Aremu 2011). In contrast, El-Adawy (2002)
water was used for cooking pulses to simulate the com- did not find any significant effect of germination and
mon domestic practice, even though its influence on Pal et al. (2016) even found increase of Ca and Fe in
mineral contents of cooked seed was predictable. germinated horse gram.
Differently, Mg levels in the unsprouted samples – raw Boiling was reported to influence the total mineral
contents of legumes in relation to variety of pulses,
and cooked – were similar, even though the Mg con-
and therefore food matrices (i.e. chelating com-
tent in tap water was 12 mg L1. In sprouted pulses,
pounds), and kind of mineral (Alajaji and El-Adawy
cooking significantly decreased the Mg contents –
2006; Wang et al. 2010). Only few data are available
27% – probably due to the mineral leaching into
about the total mineral contents in sprouted pulses,
cooking water favoured by the more porous structure
after cooking. Bains et al. (2014) found a significant
of pulses. In the latter case, a significant interaction
decrease in Fe and Ca after chickpeas germination (by
between sprouting and cooking was found (p < .05).
7% and 8%, respectively) and a further decrease in Fe,
Concerning trace minerals, varieties, sprouting and
of about 3%, after pressure cooking and microwaving.
cooking differently affected the total content of Zn
Conversely, the Zn content was not influenced by ger-
and Fe. Cooking significantly decreased the Fe content
mination, but both methods of cooking decreased the
in unsprouted pulses (35%). Conversely, in sprouted Zn content of about 7%. Unfortunately, those authors
and cooked seeds, a Fe increase in chickpeas (þ20%) did not consider boiling and did not compare
and a Fe decrease was observed in green peas (28%). unsprouted versus sprouted pulses.
As regards to the Zn content, it increased after cook- The content of phytic acid in the analysed
ing of unsprouted (þ112%) and sprouted (þ44%) unsprouted and sprouted chickpeas was more than 3-
chickpeas, probably due to the Zn uptake from tap fold higher compared to levels found in other varieties
water (0.3 mg L1), while in green peas Zn was only (Ghavidel and Prakash 2007). This high content of
minimally affected. The occurrence of different phe- phytic acid may have chelated the minerals and hin-
nomena could explain the results: in chickpea, the dered the improvement of mineral accessibility in
leaching of Fe into cooking water seems to be pre- sprouted and cooked pulses. In fact, the in vitro acces-
vented by sprouting, while germination negatively sibility of Mg and Ca in cooked chickpeas was similar,
affects the uptake of Zn from cooking water. The regardless of germination, and mineral accessibility in
observed discrepancies could be attributed to the dif- cooked sprouted green peas was even lower than in
ferent localisation of the trace mineral in seeds unsprouted ones (Table 3).
(Dalfollo Ribeiro et al. 2012) and/or the different Soaking, germination and cooking are generally
effects of germination on seed matrix (protein frac- reported as processes able to improve mineral accessi-
tion) and mineral binding compounds such as tannins, bility in pulses by a reduction of antinutritional factors
phytic acid and polyphenols. (Viadel et al. 2006; Ghavidel and Prakash 2007),
 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION 7

Table 3. Ca and Mg accessibilities in cooked chickpea and well the thickness of seed coat, represent crucial
green pea (unsprouted and sprouted). parameters for water imbibition of seeds during soak-
Chickpea Green pea ing and cooking and, consequently, on starch
Unsprouted Sprouted Unsprouted Sprouted gelatinisation (Klamczynska et al. 2001).
Ca 33.5 ± 4.4 40.0 ± 5.9 30.5 ± 4.5 19.6 ± 1.6 Sprouting under controlled conditions caused
Mg 45.8 ± 7.0 41.0 ± 2.9 43.0 ± 3.7 23.4 ± 3.7
minor and not statistically significant variations of the
Asterisks indicate statistical differences from the relative unsprouted sam-
ples (p < .05). Values are presented as mean ± SD. n ¼ 5 independent RDS and SDS percentages in both intact legumes. This
measurements and expressed as percentage of soluble mineral in relation suggests that the industrial treatment considered in
to total mineral content.
this study did not promote changes in the rate of
starch digestibility in intact pulses, likely maintaining
although conflicting results have been reported. For
a high degree of ordered structure in cell walls (Figure
example, Hemalatha et al. (2007) failed to demonstrate
1(b,f)). The cooking of whole pulses in boiling water
any increase in Zn bioavailability in chickpeas after
resulted in the swelling of the starch granules, but not
germination.
in the disruption of the cell walls or the starch gran-
To sum up, these data suggest that the accessibility
ules (Figure 2(d,h)). This phenomenon of intracellular
of minerals in pulses is the result of interactions of
gelatinisation without disruption of the starch granules
many factors – like the type of mineral, the compos-
could account for the observed low rate of starch
ition and structure of pulses and the processes – that digestion in germinated pulses. Numerous studies
cannot be easily predicted. investigated the digestibility of starch in legume flours
after germination. Benıtez et al. (2013) reported a
In vitro starch digestibility decrease of the total starch content and a significant
increase in the percentage of available starch after ger-
Although several studies on starch digestibility of
mination, ascribing these changes to the increased
unsprouted and sprouted pulses have been conducted,
a-amylase activity induced by the treatment. The
most ones have focussed on raw/cooked flours authors also suggested that the reduction of antinutri-
(Ghavidel and Prakash 2007; Uppal and Bains 2012) tional factors in the seeds after germination could pro-
or on isolated starches (Hoover et al. 2010). To date, mote the starch digestibility. The partial removal of
no information is available on the effect of sprouting phytic acid and tannins, which takes place during
on starch digestibility of whole pulses as eaten. germination, probably created a large space within
In this study, we evaluated the starch digestibility the matrix increasing the susceptibility to enzymatic
of unsprouted and sprouted pulses after cooking, by a activity and, consequently, improving the starch
well-established and extensively employed in vitro digestibility.
assay, which allowed the determination of nutritionally The findings of the present study seem quite inter-
important starch fractions, rapidly and slowly digest- esting in view of the effects of the consumption of
ible starch (RDS and SDS, respectively). By measuring sprouted legumes on glycaemic metabolism. In fact,
the susceptibility of starch to digestive enzymes, this the maintenance of the characteristics of slow digest-
assay is a commonly used method to estimate the ibility of starch, comparable to those of unsprouted
potential glycaemic response of foods (EFSA 2011). pulses, should give the germinated products a reduced
Indeed, the glycaemic response appears to be directly glycaemic impact, not unlike that typical of legumes
related to the amount of RDS and the insulin demand (www.glycemicindex.com; Sievenpiper et al. 2009;
is inversely correlated to SDS fraction (Garsetti et al. Benıtez et al. 2013). However, this potential should be
2005). As expected, a different starch susceptibility demonstrated in further in vivo studies.
(Figure 4(a)) was observed in native pulses. In chick- Besides the consumption of pulses as whole seeds,
peas, the RDS fraction was lower and the SDS one pulses are commonly used as food ingredients, thanks
was higher (20%; p < .05) than those determined in to their chemical composition, which improves the
green peas. Differences in the in vitro digestibility of nutritional quality of the finished products. Thus, we
native starches among and within species have been evaluated the starch digestibility of corresponding
attributed to the interplay of many factors, such as flours after cooking, thereby simulating a potential use
starch source, granule size, degree of crystallinity, type in porridge or baby food. In contrast to what observed
of crystalline polymorphic (A, B, or C) form, amylose/ in intact pulses, the changes in the structure of the
amylopectin ratio, molecular structure of amylopectin, seeds resulting from the grinding before cooking
amylose chain length etc. (Hoover et al. 2010). induced a significant (p < .05) increase in RDS
Moreover, differences in seed size and structure, as fraction (Figure 4(b)). These changes promoted the
8 D. ERBA ET AL.

(A) 100

90

80
A

% Starch Digesbility
70
AB a
60
ab BC
bc
50 c
40
C

30

20

10

0
unsprouted sprouted unsprouted sprouted
chickpeas chickpeas green peas green peas
RDS/AVST SDS/AVST

(B) 100 a
a a
90
a

80
% Starch Digesbility

70

60

50

40

30
A
20
A
A A
10

0
unsprouted sprouted unsprouted sprouted
chickpea flour chickpea flour green pea green pea
flour flour
RDS/AVST SDS/AVST
Figure 4. Effect of sprouting on the percentage (mean ± SD, n ¼ 6) of RDS and SDS (fractions expressed as percentages on
available starch) in cooked pulses (a) and in porridge (b). Bars with different letters (lowercase letters refer to %RDS/AVST; capital
letters refer to %SDS/AVST) are significantly different (p < .05).

digestibility of starch in both cooked chickpea and Table 4. Total antioxidant capacity in raw, soaked and cooked
pea flours, regardless of the process of germination. chickpea and pea (unsprouted and sprouted).
It is likely that grinding opened up the cell walls Chickpea Green pea
and released the starch granules, favouring their dis- Unsprouted Sprouted Unsprouted Sprouted
persion in water and the gelatinisation during cook- Raw 27.14 ± 1.57b 24.89 ± 1.29A, 27.13 ± 1.62b 24.08 ± 0.77A,
ing (Brummer et al. 2015). Although these features Soaked 40.83 ± 12.45a 16.97 ± 1.88B, 32.27 ± 5.61a 13.00 ± 2.01B,
Cooked 18.84 ± 3.72c 11.98 ± 2.82C, 13.25 ± 2.24c 8.16 ± 1.18C,
can greatly reduce the hypoglycaemic properties
Values are presented as the mean ± SD. n ¼ 6 independent measurements
assessed in intact seeds, on the other hand, they and expressed as mmol TEAC/kg dry weight. Lowercase letters indicate
may be considered as positive in view of the formu- statistical differences among the unsprouted raw, soaked and cooked
pulses, whereas capital letters indicate statistical differences among the
lation of products characterised by readily avail- sprouted raw, soaked and cooked pulses (p < .05). Asterisks indicate
able energy. statistical differences from the relative unsprouted samples (p < .05).
 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION 9

Total antioxidant capacity free and conjugated phenolics, by increasing the anti-
oxidant capacity.
The effect of the sprouting on the TAC of the analysed
Concerning the cooking process, our data showed a
pulses is shown in Table 4. Germination had a notable
significant 30% TAC reduction in both the pulses
effect in lowering the TAC of chickpeas and green
samples, compared to the raw ones. Data are in
peas. The highest decrease of TAC was observed in
accordance with Xu and Chang (2008), which reported
both chickpea and green pea soaked samples, with an
a 57–77% decrease of antioxidant capacity of cooked
about 40% decrease, raising to about 60% decrease for
green peas and chickpeas. Interestingly, the cooking
germinated cooked green peas with respect to
process had a similar influence on the TAC of
raw ones.
These results may be explained considering that sprouted and unsprouted pulses, suggesting that, des-
there is an increase of reactive oxygen substances at the pite the almost intact cell structure (Figure 1(c,d,g,h)),
beginning of the sprouting phase (Bailly et al. 2008) cooked pulses lost a part of the antioxidant potential
mainly produced by hydrogen peroxide, which is a probably due to the leaching of phenolic compounds
physiological signalling mediator, after superoxide in the cooking water and their degradation (Xu and
dismutase and catalase enzymes catalysis (Wojtyla Chang 2008).
et al. 2016). Phenolic compounds and other antioxi-
dants could have counteracted such reactive oxygen
substances with a net decrease in the antioxidant Conclusions
capacity of sprouted pulses. Amarowicz and Pegg
The availability of these partially germinated and
(2008) evidenced a similar trend for lentil samples,
stabilised pulses at an industrial scale expands the
suggesting that only after the fourth day of germin-
potential for the integration of these novel ingredients
ation an inverse trend of the antioxidant capacity
into the food and feed market and helps to meet
takes place. Moreover, an enhancement of the poly-
increasing consumer demands for natural healthy
phenol oxidase activity, which oxidises polyphenols
food products.
mainly responsible for the TAC of pulses (Sharma and Although the performed sprouting process did not
Sehgal 1992), cannot be ruled out after the sprouting
improve the mineral accessibility as expected and
and the further soaking process.
promoted a decrease of antioxidant capacity, this
On the contrary, a significant increase in TAC was
study provides evidence that sprouted chickpeas and
observed in both pulses when raw and soaked
green peas maintain their relevant nutritional traits.
unsprouted samples were compared. Results are in
Moreover, flatulence-related oligosaccharides greatly
contrast to those of Xu and Chang (2008), who
decreased and phytic acid level was significantly
showed about a 50% decrease of antioxidant
reduced. Although the lack of replication of sprouting
capacity (ORAC assay) of soaked green peas with
process of pulses might represent a weakness of the
respect to raw ones and a 4–30% decrease of soaked
present study, it demonstrated that sprouting on an
chickpeas with respect to raw ones. It should be
industrial-scale induced mild structural modification
considered that authors performed an extraction with
in chickpeas and green peas.
organic solvents, which may not have solubilised all
Overall, sprouted pulses seem to offer an excellent
the phenolics, such as the conjugated ones, highly
opportunity for developing new products aimed at
present in pulses samples (Wang et al. 2016). On the
improving the nutrient profile of products targeting
contrary, by using the method of Açar et al. (2009),
users relying on vegetarian or vegan diets.
all the phenolics contained in the seed are allowed to
react with ABTS ethanol reagent, resulting in a
higher antioxidant capacity. Our findings are also
Acknowledgements
partially in accordance with those of Segev et al.
(2011), which evidenced that the soaking process did The authors thank Mrs. Franca Criscuoli and Mr. Giovanni
not significantly affect the antioxidant capacity of Fiorillo for technical assistance, and Molino Quaglia Spa for
providing samples.
non-coloured pulses, compared to raw ones.
However, as the soaking process in our unsprouted
pulses minimally affected the integrity of the external
layers, we hypothesised that soaking water remained Disclosure statement
into the seed and may have extracted both soluble No potential conflict of interest was reported by the authors.
10 D. ERBA ET AL.

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