Nutritional, antioxidant, glycaemic index and Antihyperglycaemic properties of improved traditional plantain-based (Musa AAB) dough meal enriched with tigernut (Cyperus esculentus) and defatted soybean (Glycine max) flour for diabetic patients

Abstract

The study aimed at determining nutritional, antioxidant and blood glucose lowering potentials of improved plantain-based dough meals enriched with defatted soybean and tigernut flour. The constituted dough meals [PSB (plantain 64.46, defatted soybean 35.54%), TNS (tigernut 59.83, defatted soybean 40.17%); PTS (plantain 51.07, tigernut, 11.50, defatted soybean, 37.43%); TNT (100% tigernuts); PLT (100% plantain) and CNT (a commercial flour)] were evaluated for nutritional, antioxidant and blood glucose concentration in streptozotocin-induced diabetics rats. The improved dough meals contained appreciable amount of protein, energy value, and high in antioxidative activity than PLT. Blood glucose reducing potential of improved plantain-based dough meals (60.5–71.9%) in streptozotocin-induced diabetic rats was higher than PLT, but comparable to acarbose (anti-diabetic drug) (69%). The present study established that improved traditional plantain-based dough meals (particularly PTS) was high in essential nutrients, antioxidative activities, and blood glucose reducing potentials. Hence, the dough-meals may be suitable for diabetes management.

Full text

Nutritional, antioxidant,
glycaemic index and
Antihyperglycaemic properties
of improved traditional
plantain-based (Musa AAB)
dough meal enriched with
tigernut (Cyperus esculentus)
and defatted soybean (Glycine
max)flour for diabetic patients
Timilehin David Oluwajuyitan, Oluwole Steve Ijarotimi
∗
Department of Food Science and Technology, Federal University of Technology, Akure, Nigeria
∗
Corresponding author.
E-mail addresses: soijarotimi@gmail.com,soijarotimi@futa.edu.ng (O.S. Ijarotimi).
Abstract
The study aimed at determining nutritional, antioxidant and blood glucose lowering
potentials of improved plantain-based dough meals enriched with defatted soybean
and tigernut flour. The constituted dough meals [PSB (plantain 64.46, defatted
soybean 35.54%), TNS (tigernut 59.83, defatted soybean 40.17%); PTS (plantain
51.07, tigernut, 11.50, defatted soybean, 37.43%); TNT (100% tigernuts); PLT
(100% plantain) and CNT (a commercial flour)] were evaluated for nutritional,
antioxidant and blood glucose concentration in streptozotocin-induced diabetics
rats. The improved dough meals contained appreciable amount of protein, energy
value, and high in antioxidative activity than PLT. Blood glucose reducing
potential of improved plantain-based dough meals (60.5e71.9%) in
Received:
8 January 2019
Revised:
23 March 2019
Accepted:
8 April 2019
Cite as: Timilehin David
Oluwajuyitan,
Oluwole Steve Ijarotimi.
Nutritional, antioxidant,
glycaemic index and
Antihyperglycaemic
properties of improved
traditional plantain-based
(Musa AAB) dough meal
enriched with tigernut
(Cyperus esculentus) and
defatted soybean (Glycine
max)flour for diabetic
patients.
Heliyon 5 (2019) e01504.
doi: 10.1016/j.heliyon.2019.
e01504
https://doi.org/10.1016/j.heliyon.2019.e01504
2405-8440/Ó2019 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).

streptozotocin-induced diabetic rats was higher than PLT, but comparable to
acarbose (anti-diabetic drug) (69%). The present study established that improved
traditional plantain-based dough meals (particularly PTS) was high in essential
nutrients, antioxidative activities, and blood glucose reducing potentials. Hence,
the dough-meals may be suitable for diabetes management.
Keywords: Food science, Nutrition
1. Introduction
Diabetes mellitus (DM) is a chronic metabolic disorder characterized by increased
blood glucose levels (Ozougwu et al., 2013), due to either lack of insulin production
or inefficient in the activity of insulin (Maitra and Abbas, 2005). Diabetes Mellitus is
one of five leading causes of deaths and debilitating disease in the world (WHO,
2010;International diabetes foundation, 2012). The number of people with type 2
diabetes mellitus is increasing in every country with 80% of people with diabetes
mellitus living in low- and middle-income countries (Mufunda et al., 2006;Levitt,
2008). In Nigeria, epidemiological studies have reported that diabetes mellitus is
one of the commonest causes of admission and death in tertiary health institutions
(Osuafor and Ele, 2004;Odenigbo and Oguejiofor, 2009). Several strategies such
as regular use of different antidiabetic medications, dietary modifications and change
in lifestyle have been adopted to manage diabetes (Bantle et al., 2008;Hawley and
Gibala, 2012). These strategies, particularly using synthetic antidiabetic agents are
very costly, and also have some side effects (Prasad et al., 2014). In view of this,
efforts have focused on the development of non-toxic food-based antidiabetic
agents, that is, functional foods (Inzucchi et al., 2012;Agius, 2014).
One of the current issue in nutrition and health is to consume food products, that is,
functional foods that have the potential to provide adequate nutrient requirements,
and also to promote good health status on regular ingestion (Ju
arez-García et al.,
2006;Academy of Nutrition and Dietetics, 2017). Functional food is an emerging
field in food science due to its increasing popularity among health conscious con-
sumers. The increasing interest in therapeutic foods reflects the fact that a specific
diet or component of a diet is associated with a lower risk of certain diseases. For
instance, Odom et al. (2013) and Famakin et al. (2016) formulated plantain-based
foods fortified with legumes for the management of diabetes. Plant-based functional
foods are now getting more attention than ever before, because they have the poten-
tial of myriad benefit to the society or indeed to the entire mankind especially in the
line of nutrition, medicine and pharmacology. The medicinal values of these plant-
based foods lies in bioactive compounds like phytochemicals and proteins that pro-
duce definite physiological action on the human body (Edeoga and Gomina, 2000;
Igwe et al., 2012).
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Plantain is widely grown in the southern parts of Nigeria and other African countries.
In many parts of African countries, plantain is consumed as a cheap source of en-
ergy, and also, medically recommended for diabetic patient, due to its low glycaemic
index properties (Akubor and Ishiwu, 2013;Eleazu and Okafor, 2015). Plantain,
particularly unripe, is a good source of energy, dietary fiber, irons, potassium, and
vitamins (Randy et al., 2007;Tribess et al., 2009). Apart from being a good source
of calories and other nutrients, plantain is considered nutritionally poor, because it is
deficient in fat and protein (Odenigbo et al., 2013). Therefore, supplementation of
plantain flour with inexpensive staples, such as legumes, cereals and pulses, helps
to improve the nutritional quality of plantain products (Famakin et al., 2016). Plan-
tain flour has been successfully added to cereals to produce bread (Ju
arez-García
et al., 2006), spaghetti (Mastromatteo et al., 2014) and other food products, demon-
strating that its addition results in higher protein and resistant starch content and a
lower starch digestion rate.
Tigernut (Cyperus esculentus Lativum) is a perennial grass-like plant, and underu-
tilised tubers that produces sweet nut-like taste (Cos¸kuner et al., 2002), widely
grown in tropical and Mediterranean regions (Adejuyitan, 2011). The tuber serves
as a potentially valuable food for both human and animal in Southern Europe and
many parts of developing countries (Pascual et al., 2000). The tubers contain appre-
ciable amount of essential phytonutrients and phytochemicals, which are effective in
the treatment and prevention of many diseases like coronary heart diseases, obesity,
diabetes, and gastrointestinal diseases (Sabiu et al., 2017).
Soybean (Glycine max (L)Merrill) belongs to the family of leguminoisae and sub-
family papilionnideae. Soy protein is a major component of the food for animals
and is increasingly important in the human diet (Usman et al., 2016). Soy protein
is deficient in methionine, but high in lysine. Hence, there is a needs to complement
soybean based food products with cereals, which is high in lysine, but low in methi-
onine (Coulibaly et al., 2012). In Nigeria, Soybean is used in various food products
in different homes such as soymilk, soyogi, soy-daddawa, soy-akara, soy-gari, soy-
soup, and hot soy-drink (Coulibaly et al., 2012;Samson, 2014;Usman et al., 2016).
This study aimed at formulating and to evaluate nutritional composition, glycaemic in-
dex and antidiabetics potentials of dough meals from locally available food materials.
2. Materials and methods
2.1. Source of food materials
Tigernuts (Cyperus esculentus Lativum) yellow variety and matured, unripe fruits of
plantain were purchased from Erekesan market, Akure and Owena market, Ondo,
Nigeria, respectively.
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2.2. Processing of food materials into flour
2.2.1. Tigernut tuber flour processing
Tigernut tuber was processed into flour using the method of Oladele and Aina
(2017) with slight modification. Yellow tigernut tubers were sorted to remove un-
wanted materials like stones, pebbles and other foreign seeds, washed with double
distilled water and drained. The tubers were oven dried at 60 C for 20 h using a
hot-air oven (Plus11 Sanyo Gallenkamp PLC, Loughborough, Leicestershire,
UK), milled with a laboratory blender (Model KM 901D; Kenwood Electronic, Hert-
fordshire, UK) and passed through a 60 mm mesh sieve (British Standard) to obtain
tigernut tuber flour. The flour was packed in a plastic container, sealed and stored at
room temperature (w27 C) until analysis.
2.2.2. Plantain flour processing
The plantain flour was processed using method described by Mepba et al. (2007)
with slight modification. The plantain heads were cut into separate bunches which
were subsequently de-fingered. The fingers were washed to remove adhering soil
particles, peeled, cut into thin slices of about 2-cm thick, blanched in 1.25%
NaHS0
3
solution at 80 C for 5 min and drained. The blanched plantain slices
were oven dried at 60 C for 20 h using a hot-air oven (Plus11 Sanyo Gallenkamp
PLC, Loughborough, Leicestershire, UK), milled with a laboratory blender (Model
KM 901D; Kenwood Electronic, Hertfordshire, UK) and passed through a 60 mm
mesh sieve (British Standard) to obtain plantain flour. The flour was packed in a
plastic container, sealed and stored at room temperature (w27 C) until analysis.
2.2.3. Processing of defatted soybean flour
The defatted soybean flour was processed using method described by Ijarotimi and
Owoeye (2017). The defatted soybean was oven dried at 60 C for 20 h using a hot-
air oven (Plus11 Sanyo Gallenkamp PLC, Loughborough, Leicestershire, UK),
milled with a laboratory blender (Model KM 901D; Kenwood Electronic, Hertford-
shire, UK) and passed through a 60 mm mesh sieve (British Standard) to obtain de-
fatted soybean flour. The flour was packed in a plastic container, sealed and stored at
room temperature (w27 C) until analysis.
2.3. Formulations of food samples
The plantain, tigernut tubers and defatted soybean flour were blended with reference
to 50% recommended daily intakes (RDI) of protein (56 g/day) and fat (20 g/day) for
diabetes patient’s adult using material balance equations. This was done in order to
compensate for carbohydrate reduction, and also to maintain the energy value of the
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product for the target population. The following food combinations were thereafter
obtained, that is, PSB (plantain flour, 64.5% & defatted soybean flour, 35.5%), TNS
(tiger nut flour, 59.8% & defatted soybean flour 40.2%), PTS (plantain flour 51.1%,
tigernut flour 11.5% & defatted soybean flour 37.4%), PLT (100% plantain flour),
TNT (100% tigernut flour) and a commercial dough meal flour (control) (CNT).
2.4. Chemical analyses
2.4.1. Determination of proximate composition flour blends
Proximate compositions, that is, moisture content, ash, crude fiber, crude fat and
crude protein content of experimental food samples were determined using the stan-
dard methods (AOAC, 2012). Carbohydrate content was determined by difference as
follow:
Carbohydrate ð%Þ¼100 ð%Moisture þ%Fat þ%Ash þ% Crude fibre
þ%CrudeproteinÞ
The gross energy values of the samples were determined (MJ/kg) by using Gallen-
kamp Adiabatic bomb calorimeter (Model CBB-330-01041; UK).
2.4.2. Determination of antioxidative potential of flour blends
The scavenging effect of improved plantain-based dough meal samples on 2, 2- Di-
phenyl-1-picryhydrazyl (DPPH) was measured according to the method of Hwang
et al. (2006), metal chelating activity was determined using a modified method of
Decker and Welch (1990), nitric assay was carried out as described by scientific
methods (Kumar et al., 2008;Gupta et al., 2011), and ferric reducing power was
determined according to the method of Mau et al. (2002).
2.4.3. Glycaemic index and nutritional evaluation of dough
meals from plantain, tigernut and defatted soybean composite
flour
2.4.3.1. Experimental animals
Forty white albino rats (Rattus novergicus) of both sex weighing between 120 and
200 g was obtained from the Animal House Unit of the Department of Biochemistry,
Federal University of Technology, Akure. The rats were divided into eight groups (5
rats per group) and were kept in clean plastic cages and maintained under standard
laboratory conditions (temperature, 22 3C; photo period, 12 h natural light and
12 h dark; humidity, 40e45%) (Lawal et al., 2015). The animals were maintained on
standard animal feeds and water ad libitum.
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2.4.4. Statement of animal rights
The study protocol was approved by the Ethical Committee for Laboratory Animals
of School of Agriculture and Agricultural Technology, Akure, Nigeria (FUTA/
SAAT/2017/033). The experiments on animals were conducted in accordance with
the force laws and regulations as regards animal use and care as contained in the Ca-
nadian Council on Animal Care Guidelines and Protocol Review (CCAC, 1993).
2.4.5. Determination of glycaemic index and glycaemic load of
flour blends
2.4.5.1. Experimental animals
Twenty-five Wistar Albino rats of body weights between 140-150g were divided into
5 groups (5 rats/group), and the rats were housed individually in metabolic cages in a
climate-controlled environment with free access to feed and water. The rats were al-
lowed to acclimatize to the new environment for 7 days. After the adaptation period,
the animals were reweighed and fasted for 12 h (overnight fasting). The blood
glucose of the animals were taken at zero time from the tail vein before feeding
them with the test products and glucose (a control) in a portion size that was calcu-
lated to contain 2.0 g of available carbohydrate, which were consumed within 25 min.
After the consumption, the serum glucose levels of the animals were measured using
an automatic glucose analyzer (‘Accu-chek Active’Diabetes monitoring kit; Roche
Diagnostic, Indianapolis, USA) at 0, 30, 60, 90 and 120 min intervals. The glycaemic
response was determined as the Incremental Area under the Blood Glucose Curve
(IAUC) measured geometrically from the blood glucose concentration-time graph
ignoring area beneath the fasting level (Wolever et al., 1991).
2.4.5.2. Measurement of blood glucose response
Blood glucose curves were constructed from blood glucose values of animals at time
0, after 15, 30, 45, 60, 90 and 120 min intervals after consumption of the glucose
(control) and experimental food samples of each group. The Incremental Area Under
the Curve (IAUC) was calculated for reference food (glucose) by the trapezoidal rule
in every rats in each group separately as the sum of the surface of trapezoids between
the blood glucose curve and horizontal baseline going parallel to x-axis from the
beginning of blood glucose curve at time 0 to the point at time 120 min to reflect
the total rise in blood glucose concentration after eating the reference food (glucose).
The Incremental Area Under the Curve (IAUC) from the animals fed with the formu-
lated food samples were similarly obtained. The glycaemic Index (GI) for each diet
was calculated by ratio of Incremental Area Under two hours of blood glucose
response or Curve (IAUC) for each diet to the IAUC for glucose solution standard
according to the method of Wolever et al. (1991) using the equation below, and
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classified as follows: Low-GI ¼<55%, Medium-GI ¼56e69%, and High-GI ¼
>70% (Dona et al., 2010).
GI ¼Incremental area under 2h blood glucose curve for food samples ð2:0gÞ
Incremental area under 2h blood glucose curve for glucose ð2:0gÞX100
2.4.5.3. Calculation of glycaemic load (GL)
Glycaemic Load (GL) for each food sample was determined by the method of
Salmer
on et al. (1997). In each individual glycaemic load was calculated by tak-
ing the percentage of the food’s carbohydrate content in a typical serving food
and multiplying it by its glycaemic index (GI) value. The following formula
was used:
GL ¼Net Carbohydrate ðgÞxGI
100
Net Carbs ¼Total Carbohydrates in the food sample served. The GL was classified
as follows: Low-GL ¼<10, Medium-GL ¼11e19 and High-GL ¼>20 (Dona
et al., 2010).
2.4.6. Growth performance and protein digestibility in rats of the
dough meals
Rats in each group were fed on experimental food samples and water ad libitum
for 28 days. All diets were served to the rats in ground form after proper mixing.
Records were kept on the weight and length changes, and total food intake. The
total faeces and urine voided for the last 7 days of the trial were collected contin-
uously for 24 h. The faeces were oven dried at 60 C, while urine was preserved
in 10 mL of 10% sulfuric acid to eliminate microbial activities and prevent nitro-
genlossesbyevaporationofammonia,andthesampleswerestoredinadeep
freezer (-20 C) prior to nitrogen determination. The feed efficiency, protein effi-
ciency ratio, biological value and feed conversion ratio were computed (Agbede
and Aletor, 2003).
2.4.7. Determination of haematological and biochemical
properties of albino rats fed on dough meals from plantain,
tigernut and defatted soybean composite flour
2.4.7.1. Collection of blood sample
At the end of experimental period, that is, 28 days, the Albino rats were fasted over-
night with access to water ad libitum and sacrificed underchloroform anaesthesia.
The blood samples were collected via cardiac puncture with syringe and poured
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into heparinised and non-heparinised tubes. The non-heparinised tubes were allowed
to clot and were centrifuged at 3000 xg for 25 min to obtain the sera, and the blood
samples were stored in a deep freezer (-20 C) prior to haematological and biochem-
ical analyses at the Medical Laboratory Unit of University Health Centre, Federal
University of Technology, Akure (Agbede and Aletor, 2003;Shittu et al., 2013).
The rats’organs, that is, whole liver, two kidney and heart were excised, freed of
fat, blotted with clean tissue paper and then weighed.
2.4.7.2. Haematological and biochemical determination
The Automated Haematologic Analyzer (Sysmex,KX-21, Systmex Corporation,
Kobe,Japan) was used to analyze the haematological parameters, that is, red blood
cells (RBC), pack cell volume (PCV), haemoglobin concentration (Hbc), white
blood cells (WBC), neutrophil (NEU) and lymphocytes (LYM) using methods
described by Dacie and Lewis (2002). Mean corpuscular haemoglobin (MCH),
mean corpuscular haemoglobin concentrated (MCHC) and mean corpuscular vol-
ume (MCV) were calculated from values obtained from RBC, PCV and Haemoglo-
bin (Hbc) content (Dacie and Lewis, 2002).
The biochemical parameters were analysed using methods described by Jasper et al.
(2012). The blood sample was first centrifuged at 1,500 x gfor 10 min at ambient
temperature. The serum was then separated and used for liver function assessment
employing measurements of the enzymes aspartate aminotransferase (AST), alanine
aminotransferase (ALT) and Alkaline Phosphate (ALP). Renal function was evalu-
ated using serum concentrations of urea and creatinine. These tests were performed
using disposable kits obtained from LabtestDiagnostica S.A. (Lagoa Santa, Minas
Gerais, Brazil).
2.4.7.3. Evaluation of antidiabetic potential of formulated dough
meals from plantain, tigernut and defatted soybean composite flour
in streptozocin-induced diabetic rats
The anti-diabetic potential of dough meals from plantain, tigernut and defatted soy-
bean composite flour was determined. The baseline blood glucose levels of animals
were measured before being induced with streptozocin. Forty Wistar albino rats were
induced by single intraperitoneal injection of freshly prepared solution of streptozocin
(150 mg$kg
-1
body weight) dissolved in physiological saline in overnight fasted rats
(Abu et al., 2010). The rats were allowed to drink 5% glucose solution to avoid hy-
poglycaemic effects of the drug. Blood glucose levels in animals were measured 72 h
after streptozocin administration through tail tipping using glucometer (Accu-Chek,
Active, Roche Diagnostic’s, Indianapolis, IN, Lot No 115764) The diabetic induced
rats with serum glucose 250 mg$dL
-1
level were used (Ramdas and Balakrishnan,
2012;Parks et al., 2013), and randomly divided into 8 groups containing 5 rats per
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group. Six of the groups were fed on experimental dough meals and control sample (a
commercial dough meal flour). The remaining two groups were fed on basal diet with
and without Acabose (a synthetic antidiabetic agent). The animals were fed with the
diets for 28 days and blood glucose levels measured in the morning at regular inter-
vals by drawing blood from each rat through tail tipping and blood glucose level
measured using Accu checkÒGlucometer kit (Meiton, 2006).
2.5. Statistical analysis
Data were subjected to analysis of variance using SPSS (IBM version. 20.0, SPSS
Inc., Quarry Bay, Hong Kong)and presented as means (SEM). Comparisons be-
tween different groups was done using Analysis of Variance (ANOVA) and Dun-
can’s Multiple Range Test (DMRT). Values of p<0.05 were considered as
statistically significant as described by Yalta and Talha (2008).
3. Results and discussion
3.1. Nutrient composition and energy values of dough meals from
plantain, tigernut and defatted soybean composite flour and
control samples
The proximate composition of improved plantain-based dough meals enriched with
tigernut and defatted soybean and control sample is presented in Table 1. The range
values of crude protein and energy of the food samples were 6.20 0.03 g/100g in
PLT to 25.82 0.01 g/100g in PTS and 357.6 3.11 kcal/100g in PTS to 422.1 
2.02 kcal/100g in TNT, respectively. Whereas, the crude fibre content ranged be-
tween 1.52 0.01 g/100g in PLT and 9.17 0.10 g/100g in TNT. Comparatively,
the protein content of enriched plantain-based dough meals were higher than tradi-
tional plantain-based dough (PLT, a 100% plantain flour), commercial dough meal
Table 1. Proximate composition (g/100 g) and energy values (kcal/100g) of dough meals from plantain,
tigernut and defatted soybean and control samples.
Samples PLT TNT TNS PSB PTS CNT
Moisture 16.95
a
0.06 9.01
c
0.19 7.53
e
0.11 8.75
d
0.09 9.07
c
0.21 10.46
b
0.20
Ash 1.59
e
0.03 2.31
c
0.01 3.73
a
0.07 3.35
b
0.01 2.34
c
0.06 1.79
d
0.01
Fat 9.37
c
0.11 20.81
a
0.42 17.32
b
0.40 2.50
f
0.23 2.67
e
0.06 4.07
d
0.03
Fibre 1.52
c
0.01 9.17
a
0.07 7.984
b
0.03 1.53
c
0.04 2.52
c
0.01 0.76
d
0.01
Protein 6.20
f
0.03 7.29
e
0.10 23.93
b
0.10 15.55
c
0.22 25.82
a
0.02 13.50
d
0.07
Carbohydrate 64.37
c
0.04 51.41
e
0.33 39.51
f
0.22 68.32
b
0.19 57.58
d
0.26 69.39
a
0.30
Energy 366.61
c
0.81 422.1
a
2.02 409.6
b
2.89 357.9
d
3.11 357.6
d
4.34 368.2
c
2.03
Means (SEM) with different alphabetical superscripts in the same row are significantly different at P <0.05.
Key: PLT: 100% Plantain; TNT: 100% Tigernut; CNT: 100% Commercial dough meal; TNS: Tigernut: Defatted soybean
(59.83:40.17%); PSB: Plantain: Defatted soybean (64.46:35.54%); PTS: Plantain: Tigernut: Defatted soybean (51.07:11.50:37.43%).
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flour (CNT) (13.50 0.07 g/100g), and value obtained for plantain-tigernut com-
posite flour reported by Adegunwa et al. (2017). This observation agreed with the
reports that plantain is a good source of calories, however, it is deficient in protein
(Odenigbo et al., 2013), hence, there is a need to complement plantain flour with
inexpensive staples, such as legumes, to further improve the nutritional quality of
the plantain-based products (Famakin et al., 2016). The fibre content of food sam-
ples, particularly TNT and TNS, was significantly higher than PLT and CNT, this
observation further establishedthe nutritional qualities of the present study food sam-
ples. Recently, epidemiological studies reported on health benefits that associated
with dietary fibre intakes such as inhibiting the absorption of glucose and lipids in
small intestine, slows gastric emptying, maintaining levels of satiety and contrib-
uting towards less weight gain (James et al., 2003;Lunn and Buttriss, 2007). Soluble
fibre and resistant starch molecules are additionally fermented by bacteria in the
large intestine, producing short chain fatty acids, which help to reduce circulating
cholesterol levels (Slavin et al., 1999). For instance, consumption of different types
of dietary fibre (DF) hasbeen demonstrated to increase satiety, lower energy density,
delay of gastric emptying, and/or modulation of gastrointestinal hormones (Smith
and Tucker, 2011). Hence, dietary fibre is used to manage obesity and diabetes
(Roberfroid et al., 2010;S
anchez et al., 2012).
3.2. Antioxidant properties of dough meal from plantain, tigernut
and defatted soybean composite flour and control samples
The results of antioxidant properties of dough meals made from plantain, tigernut and
defatted soybean composite flour and control samples are presented Fig 1aee. The
ability of improved plantain-based composite flour and control flour samples to scav-
enge free radical against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay ranged from
50.64% in PSB to 57.76% in PTS, and the values were significantly (p <0.05) higher
than PLT (50.00%) and CNT (40%). The ability of improved plantain-based compos-
ite flour samples to scavenge free radical against 2, 2-Azino-bis (3-
ethylbenzthiazoline-6-sulphonic acid) (ABTS) ranged between 0.0172 and 0.0182
mg TEAC/g for PBS and PTS, respectively; and free radical scavenging potentials
was higher in plantain-based composite flour samples than CNT (0.0149 mg
TEAC/g) and PLT (42 mg TEAC/g). The ferric reducing antioxidant power
(FRAP) of the improved composite flour samples ranged from 1.66 mg AAE/g in
PSB to 11.29 mg AAE/g in PTS, while that of CNT and PLT were 0.98 mg AAE/
g and 0.60 mg AAE/g, respectively. The Fe
2þ
chelation antioxidant power of
improved flour samples ranged between 50.86 mg/mL in PSB and 63.43 mg/mL
in PTS, while PLT sample was 37.86 mg/mL and CNT was 16.86 mg/mL. The ability
of composite flour samples to prevent free hydroxyl radicals ranged from 64.57% in
PSB and 72.22% in PTS, and the values were higher in composite flour than PLT
(44.08%) and CNT (55.92%).
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The antioxidant activities and free radical scavenging abilities of the composite flour
samples were comparatively higher than control sample (CNT). This finding could
be attributed to variations in food composition and bioactive components like phy-
tochemicals, fibres and bioactive proteins, which were significantly presents in these
experimental food samples than control sample. In the last decades, several studies
have demonstrated the significant of antioxidant in diseases prevention and manage-
ments (Valabhji et al., 2001;Polidori et al., 2001). For instance, studies have
ab
cd
e
Fig. 1. (aee). Antioxidative activities of composite flour from plantain, tigernut tuber and defatted soy-
bean flour [PLT: 100% Plantain; TNT: 100% Tigernut; CNT: 100% Commercial dough meal; NS: Tiger-
nut: Defatted Soybean (59.83:40.17%); PSB: Plantain: Defatted soybean (64.46:35.54%); PTS: Plantain:
Tigernut: Defatted soybean (51.07:11.50:37.43%)].
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reported on the contributions of antioxidants in diabetes and its complications
(Pickup, 2004;Chertow, 2004).
3.3. Glycaemic index (GI) and glycaemic load (GL) of formulated
dough meal and control samples
The in-vivo blood glucose concentrations, glycaemic index and glycaemic load of
Albino Wistar rat fed on improved plantain-based dough meals, control sample
and glucose are presented in Fig. 2 (aec). The result showed that blood glucose con-
centration of experimental rat fed on improved plantain-dough meals and control
sample ranged as follows: 69e113 mg/dL in TNS, 106e130 mg/dL in PSB,
99e152 mg/dL in PTS, 89e135 mg/dL in PLT and 101e140 mg/dL in CNT, while
those rats fed on glucose ranged between (99e333 mg/dL) after food samples were
administered (Fig. 2a).
a
b
c
Fig. 2. (aec). Blood Glucose concentration, Glycaemic index (%) and Glycaemic load of Albino rats fed
on formulated dough meals and control samples [PLT: 100% Plantain; TNT: 100% Tigernut; CNT: 100%
Commercial dough meal; NS: Tigernut: Defatted Soybean (59.83:40.17%); PSB: Plantain: Defatted soy-
bean (64.46:35.54%); PTS: Plantain: Tigernut: Defatted soybean (51.07:11.50:37.43%)].
12 https://doi.org/10.1016/j.heliyon.2019.e01504
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Dietary glycaemic index (GI) is an indicator of carbohydrate quality that reflects the
effect on blood glucose, foods and usually classified into three, that is, high (>70%),
medium (56e69%) and low GI (<55) depending on the rate at which blood sugar
level rises, whereas glycaemic load is an indicator of both carbohydrate quality
and quantity in food (Wolever et al., 1991;Salmer
on et al., 1997). The glycaemic
load of experimental food samples ranged from 6.51% in TNS to 15.10 in PSB,
while that of CNT was 14.82%. For glycaemic index, the values ranged from 36.
05% in TNS to 42.95% in PLT, while CNT (a commercial dough meal flour) was
45.67%. This finding showed that the GI and GL of improved plantain-based dough
meals enriched with tigernut and defatted soybean were lower than 55% and 20%,
hence, these dough meals could be classified as low glycaemic index and load foods,
hence, the food samples may be suitable for diabetic patients. Findings have re-
ported on the benefits inherent in low GI and GL food intakes. For instance, it is
evident that low-GI foods usually reduce the risk of diabetes (Salmer
on et al.,
1997), increase insulin sensitivity (Rizkalla et al., 2004), reduce food intake and
body weight, and may reduce serum cholesterol (Warren et al., 2003;McMillan-
Price et al., 2006). Whereas, the higher the glycaemic load (GL) value, the greater
the elevation in blood glucose and in the insulinogenic effect of the food. It is
evident that long-term consumption of a diet with a relatively high GI and GL is
associated with an increased risk of type 2 diabetes and coronary heart disease
(Foster-Powell et al., 2002). Hence, the low GI and GL values that were observed
in the present study food samples may not elevate the blood glucose of their con-
sumers. This observation agreed with other scientific studies which established
that consumption of unripe plantain did not elevate the blood glucose of its con-
sumers (Menezes et al., 2010;Dan, 2011).
3.4. Nutritional evaluation of dough meals prepared from
plantain, tigernut and defatted soybean composite flour in Wistar
Albino Rat
3.4.1. Growth patterns of Albino Wistar rat fed on dough meal
and control sample
The growth pattern of Wistar Albino rat fed on the improved plantain-based dough
meals and control sample are presented in Fig. 3 aec. The nutritional status of the
experimental rats showed that the rats fed on improved plantain-based dough meals
had better growth performance when compared with those rats fed on 100% plantain,
but were comparable to those rats fed on CNT. However, the rats fed on TNS had the
highest growth performance when compared to other formulated dough meals in
terms of weight-for-age (underweight, a measure of combination of chronic and
acute malnutrition), length-for-age (LFA) (stunting, a measure of past nutritional sta-
tus) and weight-for-length (WFL) (Wasting, a measure of short-term fluctuation in
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nutritional status) indices. Nutritionally, from this present study, it could be deduced
that these improved plantain-based food samples, particular TNS, may be suitable as
a functional food to prevent malnutrition among the young children and adults.
Studies have reported on the increase of protein-energy malnutrition (PEM) in devel-
oping countries due to low nutritional qualities of traditional foods (Ikujenlola and
Adurotoye, 2014;Adepoju and Etukumoh, 2014). Hence, a low cost food that is
high in protein and energy-density such as TNS may be a desirable substitute for
expensive imported foods and low qualities local foods (Ijarotimi and Olopade,
2009;Ijarotimi and Keshinro, 2012).
3.4.2. Protein quality and relative organ weight of experimental
rat fed on formulated dough and control samples
The protein quality and relative organ weight of experimental rats fed on formulated
and control dough meals are presented in Table 2. The range values of biological
values (BV), net protein utilization (NPU) and protein efficiency ratio (PER) of
improved dough meal samples were 81.07e93.03%, 42.38e70.80% and
a
c
b
Fig. 3. aec. Weight-For-Length (wasting), Length-For-Age (stunting) and Weight-For-Age (under-
weight) of Albino rats fed on formulated dough meals and control samples. Key: [PLT: 100% Plantain;
TNT: 100% Tigernut; CNT: 100% Commercial dough meal; NS: Tigernut: Defatted Soybean
(59.83:40.17%); PSB: Plantain: Defatted soybean (64.46:35.54%); PTS: Plantain: Tigernut: Defatted soy-
bean (51.07:11.50:37.43%)].
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0.22e3.36, respectively. Comparatively, the biological value (BV), Net protein uti-
lization (NPU) and Protein efficiency ratio (PER) of the improved plantain dough
meal samples were higher than 100% plantain (PLT) (BV ¼41.46%; NPU ¼
12.43% and PER ¼2.39). However, these values agreed with the values obtained
for the control sample (CNT) (BV ¼86.25%; NPU ¼50.12% and PER ¼3.06)
and far above FAO/WHO (1989) recommendations (BV, 70%; PER, 2.7). This
observation indicates that the protein content of the formulated dough meals may
be able to support children and adults physiological needs. The relative weight or-
gans of the experimental rats, that is, kidney, liver and heart, fed on plantain-
based dough meals ranged as follows: 0.70%e1.08%, 2.56e4.93% and 0.38%e
0.68%, respectively, and these values were comparatively higher than PLT (kidney
¼0.67%; liver ¼3.38% and heart ¼0.39%) and CNT (a commercial market sample)
(kidney ¼0.70%; liver ¼2.96 and heart ¼0.35%). The bio-efficacy of these exper-
imental dough meals in rats further indicate the nutritional quality of these food sam-
ples, and also their suitability as functional food to support growth and body
maintenance. This finding agreed with other works that reported on the nutritional
qualities of foods that were formulated from the combinations of two or more
plant-based food materials (Okpala and Okoli, 2011;Ijarotimi and Keshinro, 2012).
3.4.3. Haematological and biochemical properties of Albino
Wistar rat fed on formulated dough meal and control samples
Haematological properties of Albino Wistar rat fed on plantain-based dough meals
and control sample are presented in Table 3. The range values of packed cell volume
Table 2. Nutritional quality and relative organ weight of rat feed with formulated
dough meals and control samples.
Parameters PLT TNT TNS PSB PTS CNT
Weight gained (g) 2.13 35.75 60.03 38.73 35.58 43.75
Food intake (g) 298.50 400.25 465.45 383.15 484.00 465.45
Feed Efficiency Ratio 0.13 0.09 0.13 0.01 0.07 0.09
Nitrogen Retention (%) 0.96 2.89 13.46 2.88 16.30 5.33
Biological Value (%) 41.46 81.07 89.14 83.96 93.03 86.25
Net Protein Utilization (%) 12.43 42.38 68.76 49.68 70.80 50.12
Protein Efficiency Ratio 2.39 3.36 2.28 0.22 1.20 3.06
Relative organs weight (%)
Kidney 0.67 0.77 0.70 1.08 0.71 0.70
Liver 3.38 2.65 2.56 4.93 3.44 2.96
Heart 0.39 0.42 0.38 0.68 0.39 0.35
Key: PLT: 100% Plantain; TNT: 100% Tigernut; CNT: 100% Commercial dough meal; TNS: Tigernut:
Defatted soybean (59.83:40.17%); PSB: Plantain: Defatted soybean (64.46:35.54%); PTS: Plantain: Ti-
gernut: Defatted soybean (51.07:11.50:37.43%).
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(PCV), haemoglobin concentration (Hbc) and white blood cells (Wbc) were
33.50e42.00%, 11.20e14.10 g/dL and 2.86e6.10 10
3
mm
3
, respectively. While
that of red blood cells concentrations (Rbc) and Lymphocytes were 3.67e4.65 
10
3
mm
3
and 46.0e64.0%, respectively. The high concentration of PCV, Hbc,
RBC and lymphocytes of rats fed on experimental dough meals further established
nutritional quality of these products. This finding agreed with the report of Roberts
et al. (2000) who established that diets containing quality protein and iron usually
enhance production of haemoglobin and immunity in animals). In contrary, low
PCV and Hbc that were observed in rats fed on 100% plantain dough meal could
be attributed to low protein quality of the food sample, which may lead to poor pro-
duction of haemoglobin, hence, anaemia (Osundahunsi and Aworh, 2002;Ijarotimi
and Keshinro, 2012). The range values of mean cell haemoglobin concentration
Table 3. Haematological and Biochemical properties of Albino rats fed on formulated dough meals made
from plantain, tigernut and defatted soybean flour using rats.
Parameters PLT TNT TNS PSB PTS CNT *NR
PVC% 34.50 0.86
bc
39.50 1.44
ab
33.50 0.86
c
34.33 0.33
bc
42.00 1.73
a
38.00 3.46
abc
37.6e50.6
Hb (g/dl) 11.50 0.28
b
13.20 0.51
ab
11.20 0.28
b
11.30 0.11
b
14.10 0.57
a
12.65 1.18
ab
11.5e16.1
WBC (x10
3
mm
3
) 4.65 0.37
ab
5.45 0.37
ab
4.00 0.23
bc
2.86 0.03
c
6.10 0.23
a
4.75 1.18
ab
6.6e12.6
RBC (x10
3
mm
3
) 3.77 0.10
b
4.35 0.14
ab
3.67 0.10
b
3.70 0.15
b
4.65 0.20
a
4.20 0.40
b
6.76e9.75
MCHC (g/dL) 33.3 0.00
b
33.35 0.08
ab
33.40 0.00
ab
33.26 0.06
b
33.50 0.00
a
33.25 0.08
b
28.2e34.1
MCH (pg) 30.45 0.08
ab
30.30 0.17
ab
30.70 0.11
a
29.23 0.23
b
30.35 0.08
ab
31.55 0.89
a
16.0e23.1
MCV (fl) 91.40 0.17
a
90.75 0.31
ab
91.15 0.14
ab
89.40 0.00
c
90.30 0.17
b
90.55 0.49
ab
50.0e77.8
Neutrophils (%) 48.67 0.33
a
60.00 1.15
bc
48.00 0.00
a
50.33 0.33
a
36.00 0.57
c
54.66 5.48
ab
5.3e38.1
Lymphocytes (%) 50.00 0.00
cd
60.00 1.15
ab
50.00 1.15
cd
46.00 0.57
d
64.00 0.57
a
54.67 5.47
bc
56.7e93.1
Monocytes (%) 0.00 0.00
b
0.00 0.00
b
2.00 1.00
a
3.00 0.00
a
0.00 0.00
b
2.00 1.00
a
0.00e7.7
Eosinophils (%) 1.67 0.33
a
0.00 0.00
a
0.00 0.00
b
0.00 0.00
a
0.00 0.00
a
0.00 0.00
b
0.0e3.4
Basophils (%) 0.00 0.00 0.00 0.00 0.00 0.00 0.0e0.4
Creatinine (mg/dL) 44.36 1.77
b
57.66 6.56
a
30.16 0.87
c
43.15 4.24
b
14.44 0.84
d
28.63 3.33
c
0.2e0.8
Urea (mg/dL) 4.18 0.11
b
7.61 1.91
a
2.22 0.64
bc
4.10 0.27
b
0.28 0.09
c
3.17 0.21
b
7e20
Total protein (g/dL) 2.79 0.05
b
3.18 0.05
ab
3.15 0.15
ab
3.38 0.07
a
1.28 0.13
c
2.96 0.32
ab
5.6e7.6
Albumin (g/dL) 4.24 0.17
d
6.46 0.17
b
3.81 0.26
d
5.70 0.27
c
4.31 0.10
d
8.33 0.20
a
3.8e4.8
Globulin (g/dL) 1.45 0.12
d
3.71 0.13
b
1.21 0.91
d
2.32 0.19
c
1.05 0.02
d
5.87 0.24
a
-
AST (m/L) 34.16 0.83
d
68.88 0.62
b
77.83 1.02
a
44.78 1.33
c
48.50 1.95
c
25.22 1.75
e
45.7e80.8
ALT (m/L) 136.34 18.26
a
122.14 2.65
b
117.97 0.55
c
110.95 1.31
d
115.67 1.83
c
116.56 2.52
c
17.5e30.2
ALP (m/L) 131.81 3.61
c
214.27 0.96
a
123.37 1.75
d
16.45 1.09
e
121.95 3.39
d
195.75 1.95
b
56.8e128
AST/ALT ratio 0.25
d
0.00 0.56
b
0.01 0.66
a
0.01 0.40
c
0.00 0.42
c
0.01 0.22
e
0.00 <1.0
Means (SEM) with different alphabetical superscripts in the same row are significantly different at P <0.05.PLT: 100% Plantain; TNT:
100% Tigernut; CNT: 100% Commercial dough meal; TNS: Tigernut: Defatted soybean (59.83:40.17%); PSB: Plantain: Defatted soy-
bean (64.46:35.54%); PTS: Plantain: Tigernut: Defatted soybean (51.07:11.50:37.43%). *Normal range (NR): Giannini et al. (1999);
Diana (2007).
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(MCHC), mean cell haemoglobin (MCH) and mean cell volume (MCV) of rats fed
on the experimental foods were 33.26 0.06e33.50 0.01 g/dL, 29.23 
0.23e30.70 0.11 pg and 89.40 0.01e91.40 0.17 fL, respectively, while
that of CNT were 33.25 0.08 g/dL, 31.55 0.89 pg and 90.55 0.49 fL, respec-
tively. For the neutrophils, the values ranged between 36.00 0.57 and 50.33 
0.33% for the experimental foods, while that of CNT was 54.66 1.48%. The
MCHC, MCH and MCV are a useful indices of the average Hb concentration of
the red blood cells, and low concentration of these haematological parameters in an-
imals is indicative of haemolytic anaemia, while their increased also indicate
massive intravascular haemolysis (Sridhar Prasad et al., 2018).
The total blood proteins (1.28e3.38 g/dL) and serum albumin concentrations
(3.81e5.70 g/dL) of rats fed on formulated dough meals were comparatively lower
than normal range values for serum protein (6e8 g/dL) and albumin (3.5e5.0 g/dL)
(Giannini et al., 1999;Diana, 2007), this could be as a result of the nature of protein
source, that is, from plant. It well known that the bioavailability of plant proteins is
usually lower compare to animal-based proteins, and that the concentration of
plasma proteins, especially albumin, depends on the amount of protein intakes
and its quality (Fujita et al., 1978).
The creatinine value of rat fed on the formulated dough meal ranged from 14.44 mg/
dl in PTS to 57.66 mg/dL in TNT, while that of CNT sample was 28.63 mg/dL and
PLT was 44.36 mg/dL. The Urea concentration of rats fed on experimental dough
meals ranged from 0.28 mg/dL in PTS to 7.61 mg/dL in PSB group, and were com-
parable to those rats fed on CNT (3.17 mg/dL). Comparatively, the creatinine level
of the rat fed on the formulated dough meal and control samples were within the
normal ranged of Kunitoshi et al. (1997), which implies that both the formulated
dough meal and the control samples had no negative side effect on the kidney func-
tionality. Healthy kidneys remove creatinine and urea nitrogen from blood, but the
level of creatinine and urea in blood rises with kidney failure and function, that is, the
higher the creatinine and urea value the less effective the kidney function (RusulArif
and Haider, 2014).
Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and Alkaline
Phosphatase (ALP) are all enzyme markers used to determine the functionality of
the liver. The AST values of the formulated dough meal ranged from 44.78 m/L
in PSB to 77.83 m/L in TNS, and were significantly (p <0.05) higher than 100%
plantain (PLT) (25.22 m/L) and control sample (CNT) (34.16 m/L). However, the
AST values for experimental food samples were within the normal range values
(45.70e80.50 m/L) Giannini et al. (1999);Diana (2007). For ALT, the values ranged
from 110.95 m/L in PSB to 122.14 m/L in TNT, while that of CNT was 116.56 m/L
and PLT was 136.34 m/L. Statistically, the ALT value of rat fed on 100% plantain
flour dough meals (PLT) was significantly (p <0.05) higher than those rats fed
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on formulated dough meal and control sample (CNT). The ALP values ranged from
106.45 m/L in PSB to 123 m/L in TNS, while that of PLT and CNT were 131.81 m/L
and 195.75 m/L, respectively. The AST/ALT ratios of experimental food samples
ranged between 0.25 and 0.66 for PLT and TNS, respectively, and were lower
than that of CNT (commercial dough meal flour) (0.22) and normal value (<1.0).
This observation implies that the formulated diets are suitable for consumption,
and that consumption of these food samples would not damage liver cells. Scientific
study has reported that high concentration of AST or ALT in the blood is an indica-
tion of liver function and damage (Al-Mamary et al., 2002;Aniagu et al., 2005;
Aliyu et al., 2007).
3.4.4. Trend of blood glucose concentration of streptozotocin-
induced diabetic rats fed on dough meals and control samples
The percentage of blood glucose concentration reduction of streptozotocin-induced
diabetic rat fed on formulated dough meals and control sample is presented in
Fig. 4. The percentage of blood glucose reduction in streptozotocin-induced dia-
betic rats fed on experimental food samples were in this order TNS >PSB >
PTS >TNT, that is, TNS (71.9%) had the highest blood glucose reduction; while
TNT (60.5%) had the lowest percentage reduction. In comparison, this study
showed that the experimental dough meals (TNS >PSB >PTS >TNT) had higher
potentials to lower blood glucose in streptozotocin-induced diabetic rats than 100%
plantain (41.6%) and CNT, a commercial dough meals (46.1%), but comparable to
that of BDA (an acarbose, synthetic anti-diabetic agent). This observation could be
Fig. 4. Percentage reduction of blood glucose (%) in streptozocin-induced diabetic rats fed on formu-
lated dough meals and control samples [PLT: 100% Plantain; TNT: 100% Tigernut; CNT: 100% Com-
mercial dough meal; NS: Tigernut: Defatted Soybean (59.83:40.17%); PSB: Plantain: Defatted
soybean (64.46:35.54%); PTS: Plantain: Tigernut: Defatted soybean (51.07:11.50:37.43%); BDS:
Normal Rodent Basal Diet Pellet; BDA: Normal Rodent Basal Diet Pellet administered with Acarbose
(2 mg/day)].
18 https://doi.org/10.1016/j.heliyon.2019.e01504
2405-8440/Ó2019 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
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Article Nowe01504

due to the activities of bioactive compounds such as dietary fibres, phytochemicals
and peptides that are presents in these experimental dough meals, which mighty
have inhibited the activities of apha-amylase and apha-glucosidase, delaying the
gastric emptying rate and reducing active transport of glucose across intestinal
brush border membrane (Kobayashi et al., 2000;Heilbronn et al., 2004). Also,
several studies have equally reported on antidiabetic potentials of plantain-based
food products (Shodehinde and Oboh, 2013;Iroaganachi et al., 2015;Eleazu and
Okafor, 2015).
4. Conclusion
The study established that the formulated dough meals made from plantain, tigernut
and defatted soybean contained appreciable amount of protein and energy values.
The dough meals also exhibited good free radical scavenging abilities, better nutri-
tional qualities in terms of growth performances, high biological values (>70%) and
low glycaemic index (33e50%). These experimental dough meals, especially TNS
and PTS showed significant reduction in blood glucose of diabetic-induced rats,
when compared with 100% plantain flour, control (a commercial dough meal flour)
and acarbose (anti-diabetic drug). However, out of the formulated dough meals, the
PTS (plantain, 51.07%; tigernut, 11.5% & defatted soybeans flour, 37.43%) was
rated best in terms of nutritional composition, antioxidant properties and blood
glucose reducing potential, hence, this food sample may be suitable as functional
food in managing diabetic mellitus.
Declarations
Author contribution statement
Oluwajuyitan Timilehin: Conceived and designed the experiments; Performed the
experiments; Contributed reagents, materials, analysis tools or data.
Oluwole Ijarotimi: Conceived and designed the experiments; Analyzed and inter-
preted the data; Wrote the paper.
Funding statement
This research did not receive any specific grant from funding agencies in the public,
commercial, or not-for-profit sectors.
Competing interest statement
The authors declare no conflict of interest.
19 https://doi.org/10.1016/j.heliyon.2019.e01504
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(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Article Nowe01504

Additional information
No additional information is available for this paper.
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