International Journal of Biological Macromolecules 106 (2018) 200–206

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International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Effect of domestic cooking on the starch digestibility, predicted

glycemic indices, polyphenol contents and alpha amylase
inhibitory properties of beans (Phaseolis vulgaris) and breadfruit
(Treculia africana)
E. Chinedum ∗ , S. Sanni, N. Theressa, A. Ebere
Federal University, Ndufu-Alike, Ikwo, Ebonyi State, Nigeria

a r t i c l e i n f o a b s t r a c t

Article history: The effect of processing on starch digestibility, predicted glycemic indices (pGI), polyphenol contents
Received 12 June 2017 and alpha amylase inhibitory properties of beans (Phaseolis vulgaris) and breadfruit (Treculia africana)
Received in revised form 20 July 2017 was studied. Total starch ranged from 4.3 to 68.3 g/100 g, digestible starch ranged from 4.3 to 59.2 to
Accepted 1 August 2017
65.7 g/100 g for the raw and processed legumes; Resistance starch was not detected in most of the legumes
Available online 9 August 2017
except in fried breadfruit and the starches in both the raw and processed breadfruit were more rapidly
digested than those from raw and cooked beans. Raw and processed breadfruit had higher hydrolysis
Keywords:
curves than raw and processed beans with the amylolysis level in raw breadfruit close to that of white
Type 2 diabetes
Starch digestibility
bread. Raw beans had a low glycemic index (GI); boiled beans and breadfruit had intermediate glycemic
Starch blockers indices respectively while raw and fried breadfruit had high glycemic indices. Aqueous extracts of the
food samples had weak ␣-amylase inhibition compared to acarbose. The raw and processed legumes
contained considerable amounts of dietary phenols and flavonoids. The significant correlation (r = 0.626)
between ␣-amylase inhibitory actions of the legumes versus their total phenolic contents suggests the
contribution of the phenolic compounds in these legumes to their ␣-amylase inhibitory properties.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction
The incidence of type 2 diabetes especially in the low-income
and middle-income countries (LMIC) that have the highest preva-
lence [1,2] and mortalities arising from its complications has been
on the increase despite different approaches that have been applied
to curb it. This has made it a major public health concern especially
for persons living in these regions.
Of the 387 million people or more in the world that are living
with type 2 diabetes mellitus, about 22 million reside in sub-
Saharan Africa and Nigeria being the most populated country in
Africa, has about 4 million people with diabetes living in it [3].
Many specific interventions have been carried-out in the man-
agement and prevention of diabetes and one of these interventions
is nutrition. Diet plays an important role in the development of
diabetes and the consumption of foods with known nutraceuti-
cal properties may help to control it [4]. Dietary management may
∗ Corresponding author.
E-mail address: eleazon@yahoo.com (E. Chinedum).
involve diet alone, diet with oral hypoglycemic drugs, or diet with
insulin [5,6].
One of the key targets in the management of type 2 diabetes
mellitus is the inhibition of the starch digestive enzymes such as
␣-amylase and ␣-glucosidase that are involved in the breakdown
of starch. However, the adverse effects associated with the current
drug therapies (starch blockers) that are being used in the inhibi-
tion of these enzymes have led to increased search for alternative
inhibitors from dietary or herbal sources.
Digestibility of starch is an important metabolic response that
follows consumption of a meal. Several factors have been shown to
affect the digestibility of starches in foods some of which include:
carbohydrate content of food, nutritional composition of starch
(rapidly digestible starch (RDS), slowly digestible starch (SDS) and
resistant starch (RS)), food processing techniques, dietary fiber, and
others. These factors tend to influence the spike in blood glucose
following the consumption of a meal [6,7].
In Nigerian folkloric medicine, some of the plants that have been
suggested to have useful potentials in the dietary management
of diabetes include: Bread-fruit (Treculia africana), unripe plantain
(Musa paradisiaca), cocoyam (Colocassia esculenta), yam (Dioscorea

http://dx.doi.org/10.1016/j.ijbiomac.2017.08.005
0141-8130/© 2017 Elsevier B.V. All rights reserved.
 E. Chinedum et al. / International Journal of Biological Macromolecules 106 (2018) 200–206
alata), beans (Phaseolis vulgaris), sweet potato, wheat, and others.
However, the scientific bases for the reported hypoglycemic actions
of some of these plants have not been fully investigated.
Treculia africana Decne (moraceae), commonly known as
Africana breadfruit is one of several tropical African legumes with
ethno-medicinal and nutritive values [8]. Hypoglycemic action of
the root was also demonstrated in pre-clinical studies [9].
In a study that was carried out by Ajiboye et al. [10], they
reported the hypoglycemic action of this plant in experimentally
induced diabetic rats. Furthermore, Oboh et al. [11] reported the
glycemic index (GI) (a matrix that classifies the blood glucose-
raising potential of carbohydrate foods relative to glucose or white
bread) of this fruit to be about 64.5, which placed it as an interme-
diate GI crop based on the classification of GI by Brand-Miller et al.
[12]. However, earlier study by Rincón and Padilla [13] reported
the starches in breadfruit to be easily digestible. Similarly, in a later
study, we [14] reported that consumption of cooked breadfruit by
streptozotocin diabetic rats did not ameliorate their hyperglycemic
condition. These reports on the usefulness of breadfruit in the
management of diabetes apparently conflict each other. In addi-
tion, while Rincón and Padilla [13] provided information on the
digestibility of the starch in raw breadfruit, there is no scientific
documentation regarding the effect of cooking of this fruit (which
is the form in which it is eaten in typical African diets) on the
digestibility of its starch and the activities of starch blockers that
could naturally be present in it.
Beans (Phaseolus vulgaris Leguminosae) is a legume that has
been reported to possess different nutraceutical properties [4].
Some of the important chemical constituents in the legume include:
saponins, oligosaccharides, phytates, and phenols [15].
While the raw and cooked legume from other countries of the
world have been reportedly used to improve diabetic condition
[15] due to its slow starch digestibility and phytochemical con-
stituents [4], there is paucity of such information on Nigerian beans,
especially the white variety which is the most abundant variety in
Nigeria. This is especially true as varietal difference is one of the
properties that affect the digestibility of starches in foods. Similarly,
information on the effect of processing of Nigerian white beans
on the activities of starch blockers that could also be naturally be
present in them is also limiting in literature. Moreover, studies [16]
have shown a positive correlation between the polyphenols in some
plant based foods with the inhibitory actions of these plant based
foods on these starch digestive enzymes and these polyphenols
being polar and heat labile, could also be affected by different cook-
ing processes which in essence, could affect the inhibitory actions
of these plant based foods on these starch digestive enzymes.
In line with these, the present study was designed to study the
effect of domestic cooking on the starch digestibility, predicted
glycemic indices, total polyphenol contents and alpha amylase
inhibitory properties of beans and African breadfruit.
2. Materials and methods
2.1. Chemicals
Glucose assay kit (Sigma Catalogue No: GAG020), pepsin (Sigma
Catalogue No: P6887), ␣-amylase from porcine pancreas (Sigma
Catalogue No: A3176), amyloglucosidase from Aspergillus niger
(Sigma Catalogue No: 10115), starch (Sigma Catalogue No: S2004),
3.5-dinitrosalycyclic acid (Sigma Catalogue No: D0550), maleic
acid (Sigma Catalogue No: M0375). hydroxymethyl aminomethane
(Sigma Catalogue No: 252859), Gallic acid (Sigma Catalogue:
G7384), Quercetin (Sigma Catalogue: 1592409) and Folin & Cio-
calteu’s phenol reagent (Sigma Catalogue: F9252) were products of
Sigma-Aldrich, Inc. USA. Every other chemical used was obtained
from Nigeria and was also of analytical grade.
201
2.2. Plant materials
Beans, dehulled and fried African bread fruit (Treculia africana
Decne) seeds were purchased from the meat market in Abakaliki
town, Ebonyi State, Nigeria.
2.3. Sample preparation
Approximately 50 mg of raw food samples were weighed into
different tubes. Thereafter, the samples were cooked at 100 ◦ C for
about 90 min for beans and 120 min for breadfruit in 5 ml of dis-
tilled water, using a thermostatically regulated water bath. For the
fried African bread fruit seeds, they were prepared by blanching at
a temperature of about 100 ◦ C for 10 min, drained through a plas-
tic sieve, and thereafter dehulled. The dehulled samples were then
fried by heating at a temperature of about 105 ◦ C until they turned
brown and were ready for eating. Some samples from each plant
were left unprocessed and kept as the raw (Control). Following boil-
ing of the samples, they were homogenized in the liquids in which
they were dissolved in. The raw and fried samples were homoge-
nized in a similar way as the boiled samples. Analysis was carried
out in the tubes in which the food samples were homogenized in
order to obtain data ‘as per eaten’ and to prevent retrogadation of
starches in the foods. All analysis was carried out on fresh weight
basis.
2.4. Assay of total starch
The total starch (TS) contents of the food samples were ana-
lyzed using the method of Goni et al. [17] Following preparation
of the samples as described above, the homogenized samples were
dispersed in 6 ml of 2 M KOH at room temperature for 30 min to
solubilize all the starch. Thereafter, 60 ␮l of amyloglucosidase from
Aspergillus niger (in sodium acetate buffer, pH 4.6) was added and
the whole setup was incubated for 45 min at 60 ◦ C in a water bath.
Following incubation, the food samples were centrifuged for 10 min
at 3000 × g. Subsequently, the supernatants were collected and the
concentration of glucose in each supernatant was determined using
a glucose assay kit. The amount of glucose released was measured
at 540 nm using a UV spectrophotometer against the reagent blank.
The concentration of glucose released was converted to starch by
multiplying by 0.9.
2.5. Assay of resistant starch
RS was also determined in the raw and processed samples using
the method of Goni et al. [17] Following preparation of the samples
as described above, 0.2 ml of pepsin solution (from porcine gas-
tric mucosa, 1 g in 10 ml HCl-KCl buffer (pH 1.5)) was added and
the mixture was incubated at 40 ◦ C for 1 h with constant shaking
for protein removal. Thereafter, the samples were cooled at room
temperature. Subsequently, 9 ml of 0.1 M Tris-maleate buffer, pH
6.9 was added, followed by 1 ml of a solution of ␣-amylase (from
porcine pancreas) to ensure starch hydrolysis. The setup was well
mixed and incubated at 37 ◦ C for 16 h in a water bath. Following
incubation, the samples were centrifuged at 3000 × g for 15 min and
the supernatants were discarded. The residues were collected and
moistened by the addition of 3 ml of distilled water. Subsequently,
6 ml of 2 M KOH was added to solubilize the starch, followed by
60 ␮l of amyloglucosidase (in sodium acetate buffer, pH 4.6) and the
whole setup was incubated in a water bath for 45 min at 60 ◦ C. They
were then centrifuged for 10 min at 3000 × g. The supernatants
were collected and analyzed for concentration of glucose released
using the glucose assay kit, following the procedure as described
for total starch.
202 E. Chinedum et al. / International Journal of Biological Macromolecules 106 (2018) 200–206
Table 1
Total, resistant, digestible and rapidly digestible starches (g/100 g) in raw and processed beans and breadfruit.
Groups Total starch
Raw Beans 61.50 ± 2.12 c
Boiled Beans 59.20 ± 0.28c
Raw Breadfruit 4.30 ± 0.14a
Boiled Breadfruit 39.80 ± 0.28b
Fried Breadfruit 68.30 ± 0.14d
Values are means ± SD. a–d Means with different superscript letters along each column are significantly different (P < 0.05). ND- Not detected; Rapidly digestible starch was
expressed as g/100 g of total starch.
2.6. Digestible starch
Digestible starch (DS) was calculated as the difference between
TS and RS respectively [18].
2.7. In vitro kinetics of starch hydrolysis
The method of Goni et al. [17] was used. After preparation of
the samples as described above, 10 ml of HCl-KCl buffer (pH 1.5)
was added followed by 0.2 ml of a solution containing 1 g of pepsin
from porcine gastric mucosa in 10 ml HCl-KCl buffer (pH 1.5) and
the reaction mixture was incubated for 60 min at 40 ◦ C in a water
bath. The volume was made up to 25 ml with Tris-Maleate buffer
(pH 6.9) and the pH was accurately adjusted. In order to commence
starch hydrolysis, another 5 ml of Tris-Maleate buffer containing
2.6 IU of ␣-amylase from porcine pancreas was added to each sam-
ple and the tubes were kept in a water bath at 37 ◦ C. Aliquots
(0.1 ml) were collected from each tube every 30 min from 0 to
90 min. Alpha amylase was inactivated by placing the tubes con-
taining the aliquots in a boiling water bath for 5 min. Thereafter,
1 ml of 0.4 M sodium acetate buffer (pH 4.75) and 30 ␮l of amy-
loglucosidase from Aspergillus niger were added. The samples were
incubated at 60 ◦ C for 45 min to hydrolyze the digested starch to
glucose. Subsequently, the glucose released was quantified using
the glucose assay kit.
The rate of starch digestion rate was expressed as the per-
centage of total starch hydrolyzed at different time intervals
(30–90 min) of incubation [7,19]. The 30 min hydrolysis repre-
sented the rapidly digestible starch (RDS) [19]. The hydrolysis
index (HI) was calculated from the ratio between the area under
the hydrolysis curves of the beans and breadfruit samples and
the reference sample (white bread) [7]. The predicted glycemic
indices (pGIs) of the food samples were calculated from the
percentage of TS hydrolyzed within 90 min (H90 ) and from the
HI using the equations: (i) pGIa = 39.21 + (0.803 × H90 ) and (ii)
pGIb = 39.71 + (0.549 × HI) [17,20].
2.8. Preparation of plant extracts for alpha amylase inhibition,
total phenol and flavonoid assays
Exactly 1 g of each food sample was prepared following the pro-
cedures as described for sample preparation. The homogenized
samples were filtered using Whatman filter paper and diluted to
the concentrations: 10, 20, 30, 40 and 50 mg/ml respectively for
alpha amylase inhibition and total flavonoid assays and to the con-
centrations: 2, 4, 6, 8 and 10 mg/ml respectively for total phenol
assays.
2.9. Assay of ˛-amylase inhibition
The inhibitory actions of aqueous extracts of the food samples
on ␣-amylase were studied as previously described [20]. A mea-
sured quantity (25 ␮l) of each diluted extract was mixed with 25 ␮l
of 20 mM sodium phosphate buffer (pH 6.9) containing porcine ␣-
amylase (0.5 mg/ml) (Cat. No. A3176, Sigma Aldrich Chemical Co,
Resistant starch Digestible starch Rapidly digestible starch
ND 61.50 ± 0.21c
29.00 ± 0.00b
ND 59.20 ±0.28c 25.10 ± 0.14a
ND 4.30 ± 0.14a 36.80 ± 0.00e
ND 39.80 ±0.28b 30.30 ± 0.14d
2.6 ± 0.00 65.70 ±0.14d 29.30 ± 0.14c
Germany) in different test tubes and the setup was kept at 25 ◦ C for
10 min. Following incubation of the samples, 25 ␮l of 0.5% starch in
20 mM sodium phosphate buffer was added to each test tube. The
mixtures were allowed to react at 25 ◦ C for 10 min and the reac-
tion was stopped by the addition of 50 ␮l of 96 mM dinitrosalicyclic
acid color reagent. The reaction mixtures were then incubated in a
boiling water bath for 10 min, cooled at room temperature and the
absorbance was recorded at 540 nm using a UV spectrophotometer.
The inhibitory actions of the extracts on ␣-amylase were expressed
as a percentage using the equation:
Percent inhibition = {(Absorbance of control − Absorbance
of sample)/Absorbance of control} × 100.
2.10. Assay for total phenols
The method of Singleton et al. [21] was used. To 0.1 ml of each
extract, 50 ␮l of folin ciocalteau reagent was added and the setup
was well shaken for thorough mixing. After 3 min, 0.3 ml of 20%
Na2 CO3 (in water) was added to the reaction mixture and the whole
mixture was shaken and incubated for 15 min at room temperature
after which the absorbance was read at 725 nm. Gallic acid was used
as the standard for this assay.
2.11. Assay for total flavonoids
The method of Ordon et al. [22] was used for this assay. To 0.5 ml
of each extract, 0.5 ml of 2% AlCl3 in ethanol was added and the
mixture was incubated for 1 h at room temperature. Thereafter,
the absorbance of the samples was measured at 420 nm using a
spectrophotometer. Quercetin was used as the standard flavonoid.
2.12. Statistical analysis
Data were analyzed using one-way analysis of variance. Results
were considered to be significant when p < 0.05.
3. Results and discussion
The results for the TS, RS, DS and RDS in the raw and cooked
legumes are presented in Table 1. The TS content of the raw and
processed beans samples ranged from 59.2 to 61.5 g/100 g while
that of the raw and processed breadfruit samples ranged from 4.3
to 68.3 g/100 g. Values obtained for the total starch contents of the
raw legumes were close to the values obtained by Oboh et al. [11]
and Montserrat et al. [23] for breadfruit and beans respectively,
while they were higher than the values reported by Małgorzata
et al. [24] on beans. This variation could be attributed to varietal
differences. Cooking of the raw beans did not significantly affect
(P > 0.05) their total starch contents when compared with the raw
which may be attributed to the loss of the soluble components
such as oligosaccharides, soluble dietary fiber, and others. Simi-
lar reports were given by Capriles et al. [7] on cooked A. cruentus
 E. Chinedum et al. / International Journal of Biological Macromolecules 106 (2018) 200–206
seeds. On the contrary, there was significant elevation (P < 0.05) of
the total starch contents of the boiled and fried breadfruit sam-
ples when compared with their raw. This increase could also be
attributed to the loss of some of the soluble components [24].
RS refers to starch and its degradation products that escape
digestion in the small intestine [25]. While starches of leguminous
plants have been referred to as starch with good nutritional values
owing to their relatively high contents of RS and slowly digestible
starch [24] that arises from the presence of intact tissue structures
surrounding their starch grains, surprisingly RS was not detected in
most of the raw and processed legumes except in fried breadfruit
that had a RS value of 2.6 g/100 g. While this is the first report to
the best of our knowledge, on the RS contents of processed bread-
fruit, the variation in our findings on the RS content of the raw and
processed white Nigeria beans compared with previous studies on
beans from other countries could be attributed to varietal differ-
ences. For instance, while Montserrat et al. [23] obtained a RS value
of 26.9 and 2.5 g/100 g respectively in raw and freshly cooked fava
beans, Giovana et al. [26] obtained a low RS value of 2.6 in raw com-
mon beans (Phaseolus vulgaris), Luis et al. [27] obtained a RS value
of 7.2 g/100 g in freshly cooked fava beans; Perla et al. [28] obtained
a RS value of 2.24% in cooked Ayote bean. Other factors that could
have also contributed to the results obtained for RS in the raw and
most of the processed legumes (except fried breadfruit) include:
botanical origin, starch nature (amylose/amylopectin ratio, inter-
action between starch/nutrients), presence of other components
(lipids, protein, dietary fiber, anti-nutrients and organic acids) [18]
etc.
The DS content of the raw and processed beans ranged from
59.2 to 61.5 g/100 g while that of the raw and processed breadfruit
ranged from 4.3 to 65.7 g/100 g. The non detectable levels of RS in
the raw and most of the processed beans and breadfruit samples
suggest their starches to be easily digestible.
RDS is the starch fraction that is rapidly digested and absorbed
in the duodenum and proximal regions of the small intestine lead-
ing to a rapid elevation of blood glucose. It is estimated chemically
as the starch that is converted to the constituent glucose molecules
within 20–30 min of enzyme digestion. Englyst and Hudson [29]
suggested the use of RDS expressed “as eaten” for use as a dietary
food selection parameter since its values allow for comparison
among foods and among food portions. Results obtained for RDS
(reported as eaten) showed that 25.10 g/100 g of the starch in the
boiled beans were rapidly digestible and results obtained were sig-
nificantly lower (P < 0.05) than that of the raw beans that gave a RDS
value of 29 g/100 g. Similarly, 30.30 g/100 g of the starch in boiled
breadfruit and 29.3 g/100 g of the starch in the fried breadfruit were
rapidly digestible and results obtained were also significantly lower
(P < 0.05) than that of the raw breadfruit that gave a RDS value of
36.8 g/100 g. In addition, the findings of this study revealed that the
starches in both the raw and processed breadfruit samples were
more rapidly digested than those from the raw and cooked beans.
Fig. 1 shows the in vitro starch hydrolysis curves of the raw and
processed food samples. The digestion rate for the raw and pro-
cessed bean samples increased more slowly within the first 30 min
of amylolysis than most of the breadfruit samples and white bread
except the fried breadfruit whose digestion rate within the first
30 min was similar to raw beans. Indeed, starch digestion rate is
useful for predicting postprandial glycemic response, with rapidly
digested starches increasing glycemia more quickly [30]. Further-
more, both the raw and processed breadfruit samples exhibited
higher hydrolysis curves than the raw and processed beans samples
(with the hydrolysis curve of raw breadfruit close to that of white
bread except at 60 min hydrolysis), suggesting that both forms of
breadfruit produced higher glycemic responses under in vitro con-
ditions compared with the raw and cooked beans.
203
Fig. 1. In vitro starch digestibility of raw and processed beans and bread fruit.
Our findings on the excellent digestibility of the starch from
breadfruit is consistent with earlier studies by Rincón and Padilla
[13] who also reported high digestibility of the starch from bread-
fruit and which increased digestibility they suggested to be that the
amylopectin in the plant’s starch might contain a larger amount of
longer branched chains judging from the results of their amylo-
viscographic study. In addition, findings from this study also
suggest the non-detectable levels of RS in the plant to be another
plausible explanation for the high digestibility of the starch in
African breadfruit. This high digestibility of the starch from African
breadfruit could be one explanation for its inability to ameliorate
the blood glucose levels of streptozotocin induced diabetic rats as
previously reported [14].
The HI and pGIs of the raw and processed legumes are shown in
Table 2. The estimated glycemic indices, that is GI1 from the HI and
GI 2 from the percentage of hydrolysis within 90 min, were well
correlated with each other (r = 0.99) and the average was taken as
the pGIs for the food samples. The HI of all the raw and processed
legumes ranged from 32.57 to 83.81 while their pGIs ranged from
55.23 to 83.97. HI can be used to predict the in vivo effects of starchy
foods and it expresses the digestibility of the starch in foods in
relation to the digestibility of starch in a given standard such as
white bread [7].
To predict the in vivo effects of the different processing methods,
a HI based predicted glycemic index (pGI) was determined. Results
obtained indicate that there was significant elevation (P < 0.05) in
the hydrolysis indices of boiled beans compared with the raw but
significant reduction (P < 0.05) in the hydrolysis indices of boiled
and fried breadfruit compared with the raw.
Based on GI, foods can be classified into high (≥70), intermediate
(56–69) and low-GI (≤55) [17,20]. In line with this classification,
the findings of this study revealed that raw beans had a low GI;
boiled beans and breadfruit had intermediate glycemic indices
(GIs) respectively while raw and fried breadfruit had high glycemic
indices.
It is noteworthy that despite the non-detectable levels of RS in
the raw beans samples, it had a low glycemic index. In addition to
starch, legumes also contain high amounts of dietary fiber in a form
that gives their cell walls high resistance to disintegration [27]. It is
possible that some quantities of the starch in the raw beans were
present as slowly digestible starch or as components of soluble fiber
while boiling could have increased their digestibility which made
the boiled form to have a higher digestibility of its starch than the
raw.
There was a significant positive correlation between the pGIs of
the food samples versus RDS (r = 0.753; p = 0.01) (Table 2), suggest-
ing the contribution of RDS to starch digestion and glycemic indices
of foods.
204 E. Chinedum et al. / International Journal of Biological Macromolecules 106 (2018) 200–206

Table 2
Predicted glycemic indices of raw and cooked beans and breadfruit; Correlation of pGI versus RDS.

Groups pGI (H90 ) HI pGI (HI) Mean pGI RDS

Raw Beans 52.86 ± 2.94 a

32.57 ± 3.88a
57.59 ± 3.41a
55.23 0.753**
Boiled Beans 59.61 ± 0.66b 48.38 ± 4.30b 62.94 ± 2.50b 61.28
Raw Breadfruit 74.54 ± 0.65d 83.81 ± 0.30e 85.72 ± 1.00e 80.13
Boiled Breadfruit 62.98 ± 1.44bc 56.38 ± 2.10c 70.66 ± 1.82c 66.82
Fried Breadfruit 65.87 ± 1.71c 63.24± 1.32d 74.43 ± 0.52d 70.15
pGI

Values are means ± SD. a–d Means with different superscript letters along each column are significantly different (P < 0.05). **Correlation is significant at p = 0.01 level; HI-
Hydrolysis index; pGI-Predicted glycemic indices of raw and cooked beans and breadfruit were calculated as the average of pGI (H90 ) and pGI (HI).

Table 3 Table 4
␣-amylase inhibitory properties (%) of aqueous extracts of raw and processed beans Polyphenol contents of aqueous extracts of raw and processed beans and breadfruit.
and breadfruit.
Groups Total phenols Total flavonoid
Groups ␣-amylase inhibition (g GAE/100 g) (g QE/100 g)

Raw Beans 26.04 ± 7.61b Raw Beans 0.80 ± 0.08b 0.51 ± 0.02c
Boiled Beans 6.48 ± 8.52a Boiled Beans 0.83 ± 0.05b 0.36 ± 0.04b
Raw Breadfruit 26.86 ± 8.93b Raw Breadfruit 1.78 ± 0.8a 0.23 ± 0.00a
Boiled Breadfruit 25.23 ± 6.93b Boiled Breadfruit 2.11 ± 0.07c 0.87 ± 0.08d
Fried Breadfruit 23.78 ± 13.54b Fried Breadfruit 1.74 ± 0.05a 0.21 ± 0.02a
Acarbose 69.08 ± 21.40c
Values are means ± SD. a–d Means with different superscript letters along each col-
Values are means ± SD. a–d Means with different superscript letters along each col- umn are significantly different (P < 0.05). GAE-Gallic acid equivalence; QE-Quercetin
umn are significantly different (P < 0.05). equivalence.

range of concentration of 2–10 mg/ml), the inhibitory activities

Fig. 2. ␣-Amylase inhibitory properties (%) of aqueous extracts of raw and processed
beans and breadfruit.
Table 3 shows the percentage of inhibition of the aqueous
extracts of the food samples investigated in this study on alpha
amylase activity in vitro while the graphical representation of their
percentages of inhibition is shown in Fig. 2.
Alpha amylase is one of the key enzymes in the human body
that is responsible for the breakdown of starch to simple sug-
ars. The enzyme hydrolyzes complex polysaccharides, producing
oligosaccharides and disaccharides which are further hydrolyzed
by ␣-glycosidases to monosaccharides that are then absorbed
through the small intestines into the hepatic portal vein, increasing
postprandial glucose levels. Amylase inhibitors are also known as
starch blockers as they prevent dietary starch from being absorbed
by the body, thereby decreasing the spikes in postprandial blood
glucose levels. Slowing the digestion and breakdown of starch
through inhibition of the activities of these digestive enzymes could
be a beneficial approach in glycemic index control in people with
type 2 diabetes [31] and for borderline patients.
Findings from this study revealed that the aqueous extracts of
the food samples demonstrated varying degrees of inhibition of
alpha amylase that ranged from 6.48 to 26.46% (over a concen-
tration range of 10–50 mg/ml) and the percentage of inhibition
of some of the extracts were not dose dependent. Furthermore,
when compared with the inhibitory action of the standard anti-
diabetic drug-acarbose (which recorded 69.08% inhibition over a
demonstrated by these extracts were found to be quite weak.
The results of the phenolic and flavonoid contents of the foods
samples are shown in Table 4. The total phenolic contents of the
raw and processed bean samples ranged from 0.8 to 0.83 g Gal-
lic acid equivalence (GAE)/100 g while the total phenolic contents
of the raw and processed breadfruit samples ranged from 1.74 to
2.11 g GAE/100 g. Similarly, the total flavonoid contents of the raw
and processed bean samples ranged from 0.36 to 0.51 g Quercetin
equivalence (GAE)/100 g while that of the raw and processed bread-
fruit samples ranged from 0.21 to 0.87 g QE/100 g.
Polyphenols are phytochemicals found in plant foods that pos-
sess antioxidant properties. These polyphenols could be classified
into 5 groups namely: phenolic acids, stilbenes, lignans, flavonoids
and curcuminoids [32]. Phenolics and flavonoids have received con-
siderable attention because of their potential antioxidant activities
and effects on carbohydrate metabolism involving the inhibition
of ␣-amylase and ␣-glucosidase, thereby slowing down the diges-
tion of dietary carbohydrates to glucose [16]. These phenolic acids
and flavonoids exert inhibitory actions on alpha amylase by bind-
ing covalently to it, changing its activity as a result of their ability
to form quinones or lactones that react with nucleophilic groups
on the enzyme molecule [33].
As shown in Table 4, there were no significant differences
(P > 0.05) in the total phenolic contents of the boiled beans and
fried breadfruit compared with their raw which suggests that boil-
ing of beans or frying of breadfruit did not significantly affect the
phenolic compounds in them. On the other hand, there were sig-
nificant increases (P < 0.05) in the total phenolic contents of the
boiled breadfruit compared with the raw, suggesting that the boil-
ing water could have enhanced the extraction of bound phenolics
from the food matrix. Cooking has been reported to soften the
cell wall and other components of cells, such as vacuoles and the
apoplast, releasing phenolic compounds. Another factor that might
have also contributed to the increase in the phenolic content of
boiled breadfruit is the decomposition of phenolic compounds that
are linked to fibers (cellulose and pectin), or even the disruption
of the bonds between phenols and sugars [21]. Similar observation
was made in Solanum paniculatum fruit [34].
Results presented in Table 4 also revealed that there were sig-
nificant decreases (P < 0.05) in the flavonoid contents of the boiled
 E. Chinedum et al. / International Journal of Biological Macromolecules 106 (2018) 200–206
Table 5
Correlation between ␣-amylase inhibition versus polyphenol contents of raw and
processed beans and breadfruit.
␣-Amylase inhibition
Total phenols 0.626*
Total flavonoids 0.200
*
Correlation is significant at P < 0.05.
beans when compared with the raw. We assume that this decrease
could be due to their breakdown or these flavonoids being polar
could have leaked into the boiling water.
On the other hand, boiling of breadfruit significantly increased
(P < 0.05) its flavonoid contents.
The increase in the content of flavonoids could be associated
with high-temperature cooking time, which promoted denatura-
tion of the fruit matrix and increased the extractability of these
compounds [34]. Similar observations were also made in Solanum
paniculatum fruit [34].
The non significant differences (P > 0.05) in the flavonoid con-
tents of the fried breadfruit compared with the raw suggests that
the frying process retained the flavonoid content of the breadfruit.
Correlation analysis carried out revealed that there was a
significant positive association (r = 0.626; p = 0.05) between the ␣-
amylase inhibitory actions of the aqueous extracts of the food
samples versus their total phenolic contents but a weak positive
correlation (r = 0.200) between the ␣-amylase inhibitory actions of
the aqueous extracts of the food samples versus their total flavonoid
contents (Table 5). The significant correlation obtained between
the percentage inhibitions of the aqueous extracts of these food
samples versus their ␣-amylase inhibitory properties suggests the
contribution of the phenolic compounds in these legumes to their
␣-amylase inhibitory properties. These phenolic compounds have
been reported to be mild inhibitors of ␣-amylase activities [35].
While the mild inhibitory actions demonstrated by these legumes
on ␣-amylase activity could be desirable (as one of the reasons for
increased search for alternative medicines to the current synthetic
starch blockers is that these starch blockers exert anti-diabetic
actions through excessive inhibition of pancreatic ␣-amylase activ-
ity, resulting in the bacterial digestion of undigested complex
carbohydrates in the colon [36] with resultant gastrointestinal side
effects), we cannot draw a conclusion as per inhibitory actions of
these food samples on starch digestive enzymes as we did not inves-
tigate their inhibitory actions on ␣-glucosidase that further acts on
the metabolites from ␣-amylase digestion, converting them into
products absorbable in the small intestine as earlier stated. More-
over, inhibitory activities demonstrated in in vitro studies do not
always correlate with in vivo activity [37]. Therefore preclinical and
clinical studies will need to be carried out to validate our findings.
4. Conclusion
The study showed that the raw form of African breadfruit pos-
sessed high glycemic indices which could be attributed to the high
digestibility of its starch while the boiled and fried forms possessed
intermediate glycemic indices. On the contrary, both raw and boiled
forms of the Nigerian white beans possessed low glycemic indices.
Finally, the raw and processed breadfruit and beans were found to
exert mild inhibition of ␣-amylase activities under in vitro condi-
tions but which would need to be confirmed using clinical studies.
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