J Food Sci Technol
https://doi.org/10.1007/s13197-020-04558-4
ORIGINAL ARTICLE
Extrusion of fermented rice-black gram flour for development
of functional snacks: Characterization, optimization and sensory
analysis
Poonam Rani1 • Amit Kumar1 • Soumya Ranjan Purohit2,3 • Pavuluri Srinivasa Rao1
Revised: 4 May 2020 / Accepted: 28 May 2020
 Association of Food Scientists & Technologists (India) 2020
Abstract Considering the health benefits of fermentation,
extrusion of fermented rice black gram mix flour can lead
to developing nutritionally rich expanded snack. Hence,
present study details the effect of various extrusion process
variables, barrel temperature (100, 110, 120 C), screw
speed (250, 300, 350 rpm) and die opening diameter (3,
3.5 mm) on physicochemical and bioactive properties of
extrudates from fermented rice-black gram flour (Rice
flour: black gram flour = 3:1). Results indicated that tem-
perature rise led to the production of a more expanded
product with higher water solubility index (WSI) and lower
water absorption index (WAI). Higher puffing, WSI, and
bioactive content were observed in 3 mm die opening
extrudates. Interestingly, extrusion cooking of fermented
flour has shown a remarkable increment in phenolic con-
tent and antioxidant activity from 63.47 up to 210.3 and
7.28 up to 13.889 mg GAE/100 g, respectively. However,
rise in barrel temperature showed a negative impact on
bioactive attributes. Further, the optimal conditions deter-
mined by numerical optimization method for development
of fermented flour-based extrudates having superior func-
tional and enhanced bioactive properties were found to be
100 C temperature, 289 rpm of screw and 3 mm die
& Poonam Rani
poonam.fpe@gmail.com
1
Agricultural and Food Engineering Department, Indian
Institute of Technology Kharagpur, Kharagpur,
West Bengal 721302, India
2 1.4% ash (Sawant et al. 2013). Rice-based extruded snacks
Department of Chemical Engineering, Vignan’s Foundation
for Science Technology and Research, Guntur,
Andhra Pradesh 522213, India
3
Amity Institute of Food Technology, Amity University Uttar
Pradesh, Noida, Uttar Pradesh, India
diameter. Principal component analysis indicated that bulk
density-moisture content and phenolic content-antioxidant
activity were highly positively correlated, while expansion
ratio-bulk density was negatively correlated. The
microstructure of extrudates showed continuous and com-
pact structure lacking starch granules. The fuzzy logic
analysis revealed that fermented flour-based extruded
product had superior overall acceptability over unfer-
mented flour extrudates.
Keywords Extrusion  Rice-black gram  Fermentation 
Microstructure  Principal component analysis  Fuzzy
logic
Introduction
Extrusion cooking has been developed extensively for the
continuous processing of traditional and new product
development. During extrusion processing, raw food
material is exposed to thermal and shear energies that
transform it into modified intermediate and finished prod-
ucts (Ilo et al. 1996). It is considered as a high-temperature
short time (HTST) process that prevents the degradation of
the essential temperature-sensitive nutrients.
Ready to eat extruded snacks product are majorly pre-
pared basically with a high starch cereal grain like rice as
the primary raw material so that high expansion can be
achieved. Rice (Orzya sativum) contains about 7.37%
protein, 2.2% fat, 64.3% carbohydrates, 0.8% fibre, and
products are found to have good mouthfeel and texture;
however, they are deficient in other nutrients like proteins
and other vital nutrients. Ever-increasing consumption of
snacks has resulted in an increasing demand for healthy
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and value-added snacks products. Incorporation of the high
pulse such as black gram in the formulation while pro-
cessing through extrusion could result in the development
of nutritionally rich product snacks. Black gram (Phaseolus
mungo) lentil, contain high protein (25%), high fibre (18%)
and is one of the most abundant sources of the vitamin B
(Priyanka and Sudesh 2015). It has been reported that
fermentation of cereals and pulses provide high nutritive
value, better organoleptic characteristics, and enriches the
food with a substantial amount of proteins and vitamins
(Padmashree et al. 2009). Along with the essential nutri-
ents, bioactive compounds of food have a vital role to play
in protecting human against many chronic and degenera-
tive diseases (Brennan et al. 2011). Fermentation is known
for improving the bioactive properties of food products. Lu
et al. (2005) carried out the natural fermentation of rice
flour at 25 C for a duration of 144 h. The fermented dried
rice was used for the development of rice noodles, which
presented favorable chewy mouthfeel and better sensory
acceptability than noodles with unfermented rice. Fer-
mentation of brown rice flour for 6.26 h using 1% yeast led
to a significant increase in the protein, total ash, insoluble
fibre, soluble fibre, minerals, phenolics, antioxidants,
riboflavin, resistant starch level (Ilowefah et al. 2014). One
of the study also revealed that the fermentation of black
gram for 45 h caused 13.3% hydrolysis of phytate phos-
phorous, which helped in increasing of the bioavailability
of multivalent cations such as Ca??, Mg??, Zn??,
Fe??, and Fe??? (Reddy and Salunkhe 1980). More-
over, fermentation also contributes in improving the
availability of amino acids (Padhye and Salunkhe 1978).
Considering these health benefits, extrusion of fermented
rice black gram mix flour can be an effective way to pro-
duce nutritionally rich expanded snack with higher con-
sumer acceptability. However, a very few studies have
been carried out on the combination of extrusion and fer-
mentation for development of food products using the
fermented maize-soybean blend (Ojokoh et al. 2015) and
fermented maize-finger millet blend (Onyango et al. 2004).
Study carried out by Rani et al. (2018) showed that fer-
mentation significantly improved the bioactive properties,
protein content of the rice-black gram flour and extruded
ready to eat snacks produced using fermented flour shown
good sensory acceptability.
Along with the raw materials, extruder operating con-
ditions also affect the physicochemical properties of the
puffed extruded snacks. As reported in many studies, the
critical quality characteristics of expanded ready to eat
products such as expansion, bulk density, water absorption
and water solubility index as well as bioactive properties
are greatly influenced by the moisture content of raw
material, temperature and pressure developed in the
extruder and screw speed along with die diameter (Samyor
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J Food Sci Technol
et al. 2018; Singh et al. 2019). Hence, analysis of the effect
of process variables followed by extrusion process opti-
mization is an important step during the development of
any food product. There has been no study on evaluation of
extrusion operating parameters on physicochemical prop-
erties of puffed snack prepared from fermented rice-black
gram mix.
Hence, the purpose of the current work was to investi-
gate the effect of extrusion processing conditions on the
physicochemical and bioactive properties of a developed
fermented rice-black gram flour-based extruded product
and further optimizing the extrusion process. In addition to
it, the fuzzy logic approach has been used for sensory
analysis of the developed product to ensure its
acceptability.
Material and methods
Material procurement and sample preparation
Polished rice flour, black gram and dry yeast were pur-
chased from the local market, IIT Kharagpur. The black
gram was further ground into very fine flour. The polished
rice flour and black gram flour were mixed in the ratio 3:1.
The dough was prepared using the flour mix with the yeast
suspension which was made with 3% yeast, 4% sugar in
85% water to allow the growth and metabolic activities of
the yeast during solid-state fermentation. Yeasts are com-
monly used for the solid-state fermentation due to their
characteristics ability to grow in low moisture content
environment (Martins et al. 2011). Moreover, yeast pro-
duces higher secondary metabolites and health-promoting
compounds. Beside, fermentation is faster as compared to
natural fermentation, and hence yeast fermentation is pre-
ferred (Rani et al. 2018). The prepared dough was allowed
to ferment under controlled temperature (35 C) environ-
ment inside a BOD incubator for 6 h. For the production of
expanded/puffed material using the extrusion, the moisture
content of the raw material should be 10–15%. Therefore,
the fermented dough material was spread in a thin layer of
0.5 cm on an aluminium tray and dried in the hot air oven
dryer maintained at temperature 45 C (Ojokoh et al.
2015). The material was dried till it reached the final
moisture content of 10% wb, which was attained after a
drying of about 7 h. The dried material was ground and
passed through standard ASTM50 sieve to achieve flour of
particle size less than 0.3 mm. Before the extrusion oper-
ation, the moisture of the prepared fermented flour was
again checked and adjusted to predetermined level of
10% wb by addition of calculated amount of water or
removal of extra water through drying.
J Food Sci Technol
Extrusion processing
A laboratory co-rotating twin-screw extruder (Make:
BTPL; Model: EB-10) having L/D ratio of 14.4:1 was used
for conducting the experiments. The main drive was pro-
vided with 10 HP motor (400 V, 3 phase, 50 Hz). The
barrel consists of two parallel co-rotating screws of inter-
meshing types, driven by the drive assembly. The die
opening size, temperature of the die zone, and screw speed
of the extruder were varied according to the experimental
design. The fermented flour was introduced to the twin-
screw feeder (L/D = 4.5:1) of the extruder at a mass flow
rate of 11 kg/h. The puffed extrudates sample were col-
lected, stored in the polyethylene bags and used for further
analysis work.
Experimental design
A full factorial experimental design was selected for opti-
mization of the extrusion process for improved physical
and bioactive properties. The experimental design was
consisted of 3 independent variables; barrel temperature
(100, 110, 120 C), screw speed of extruder (250, 300,
350 C) and diameter of die opening (3 mm, 3.5 mm),
providing a set of 18 experiments. The levels of each
variable were decided based on literature and preliminary
experiments for smooth extrusion cooking. Dependent
variables were moisture content of the extrudates, expan-
sion ratio, bulk density, hydration properties, texture profile
analysis, bioactive properties. All the experiments were
done in triplicates and the average of the replicates has
been reported.
The optimization of the extrusion processing conditions
was carried out according to Rani et al. (2018). Each
response was transformed into dimensionless desirability
function (di), ranges between 0 and 1. Maximization of any
response was denoted by a desirability function (G), which
was calculated by combining the individual desirability (di)
values of all the variables.
G ¼ ðd1  d2  . . .. . .. . . dn Þ1=n ð1Þ
where di indicates the desirability of individual ith response
and n is the number of responses.
Experimental methodology
Moisture content
Moisture content (MC) of the cylindrical extruded samples
(just after emerging from the die) was estimated by the
convection drying method (Rani et al. 2018).
Expansion ratio
Expansion is the most important functional parameter of
the extruded product, which is also related to the product’s
water solubility, water absorption, crispiness, and hardness.
Expansion ratio (ER) was determined using Eq. 2. The
diameter of the extrudate was measured at 10 different
random positions of the extrudates by using digital Ver-
nier’s calliper (Rathod and Annapure 2017).
De
ER ¼ ð2Þ
Dd
where ER: expansion ratio; De: cross-sectional diameter of
extrudates (m); Dd: diameter of circular die (m).
Bulk density
Bulk density of the extrudate was determined by measuring
the actual dimensions of the extrudate and its weight. The
measured value of diameter, length and dimension of a
piece of extrudate were used to calculate the density using
the Eq. 3, assuming the extrudates of cylindrical shape
(Rathod and Annapure 2017).
4W
BD ¼ ð3Þ
pD2e l
where BD: bulk density of the extrudate; W: weight of the
extrudate (kg); De: diameter of the extrudate (m); l: length
of the extrudate (m).
Ten pieces of extrudates were randomly selected and
bulk density of the each extrudate was determined, and the
average was taken.
Water absorption index (WAI) and water solubility index
(WSI)
Hydration properties (WAI and WSI) of the extruded
samples were determined using the method explained by
Kaur et al. (2014). Briefly, 2 g of ground dry sample was
suspended and mixed with 20 ml distilled water followed
by stirring for 30 min. Further, the solution was centrifuged
at 8000 rpm for 15 min and further, separating and drying
the supernatant and sediment. The WAI (g/g) and WSI (%)
were estimated by the following given Eqs. 4 and 5.
Wws
WAI ¼ ð4Þ
Wds
Wsolid; supernatant
WSI ¼ ð5Þ
Wdry; sample
where Wws: weight of wet sediment; Wds: weight of dry
sediment; Wsolid,supernatant: weight of solid in supernatant,
Wdry,sample: weight of the dry sample.
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Texture Analysis
The textural properties of the puffed extrudates were measured
using a textural analyzer (Make: Stable micro system Texture
Technology corp., UK; Model: TA-XT2i texture analyzer).
Texture analyzer was fitted with a stainless-steel probe of
6 mm diameter, and extrudates were punctured at a pre-test
speed of 10 mm/s, test speed and post-test speed of 0.5 mm/s,
up to 30% of the extrudate height with a load cell of 5 kg. The
crispiness was calculated as per the methods described by
Sawant et al. (2013) and using the given equations.
n then transferred to the microscope for examination at dif-
Nsr ¼ ð6Þ
d ferent magnification.
s
Fcr ¼ ð7Þ
d
Fcr
Crispinesswork ¼ ð8Þ
Nsr
where Nsr: spatial frequency of rupture (mm-1); Fcr:
average crushing force (N); n: number of peaks in force–
time curve; s: area under the curve (mm2); d: probe travel
distance (mm).
Total phenolic content
Ethanolic extracts of the samples used for estimation of
TPC and AA was prepared using the method described by
Rani et al. (2018). Determination of the TPC was done
using Folin Ciocalteu Reagent according to the method
explained by Rani et al. (2018).
Antioxidant activity
Total antioxidants activity (AA) was determined using
DPPH (2,2-diphenyl-1-picrylhydrazyl) assay according to
the method described by Rani et al. (2018).
Sensory evaluation of the product
Sensory evaluation was done for extrudates prepared from
unfermented and fermented flour at optimized extrusion
conditions. Product was developed with added Maggie
masala (2.5%) in the feed flour and then coating the dried
extrudates with commercial chat masala (5%) with the help
of soybean oil (5%). A semi-trained panel of 16 members
(aged 22 to 30 years from non-smokers category) was
selected for the test. Judges were asked to mark the pro-
duct’s colour, taste, texture, mouthfeel, aftertaste and
overall acceptability according to fuzzy scale with 5-point
hedonic scale (1-poor, 2-fair, 3-good, 4-very good, 5-ex-
cellent) (Fadlallah et al. 2010). The set of observations
were analyzed using the fuzzy analysis, as explained by
Das (2005) and Debjani et al. (2013).
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Scanning electron microscopy
The microstructure of the puffed extrudates was examined
under the scanning electron microscopy (EVO 60, Carl
ZEISS SMT, Germany) at an accelerating voltage of 30 kV
under vacuum. The extruded samples were prepared for
microstructure analysis by cutting the extrudates into
rectangular pieces of thickness (1–2 mm) using a sharp
blade without damaging the structure. This was done to
expose the sample surface to an electron beam (Rani et al.
2019). Further, samples were coated with gold coating and
Statistical analysis
All the chemical analyses were performed in triplicate, and
average values are presented with respective standard
deviation. Analysis of variance (ANOVA) and numerical
optimization was done using the Design Expert V7.1,
states, U.S. software. Further, multivariate unsupervised
method of chemometry, principal component analysis
(PCA) and clustering methods were used to study the
similarities in the samples using R platform software
(Granato et al. 2018). Significant principal components
(PCs) are selected based on Kaiser criterion, i.e. eigenvalue
higher than 1 (Kaiser 1960). Before performing PCA, the
Bartlett test of sphericity was conducted, and it was found
that variables are significantly correlated with p-
value \ 0.0001.
Results and discussions
Influence of the extrusion process parameters on various
physicochemical and bioactive components has been
investigated and reported in the following sub-sections.
Statistical significance of the independent terms was
examined by the Analysis of Variance (ANOVA) for each
response. The ANOVA table (Table 1) presents the Fisher
value (F-value) and higher is the F value, more significant
is the term, or it can be said the terms are significant at
p \ 0.05.
Effect of extrusion processing on physicochemical
and bioactive properties of fermented flour-based
puffed product.
Moisture content of the extrudates
The moisture content of the extruded product was found to
be in the range of 2.84–5.89% db, which made the devel-
oped extruded products self-stable for storage. Moisture
J Food Sci Technol

Table 1 ANOVA
Effect MC ER BD TPC AA HD CW WSI WAI
table showing F values for
extrusion process of fermented DD (A) 1.77 30.08* 3.21 2.36 0.11 0.51 1.39 0.81 10.62
flour
Temp (B) 0.03 0.03 1.87 40.51* 2.41 0.51 2.70 0.08 0.96
SS (C) 6.28* 11.93* 8.92* 0.80 0.87 0.45 0.05 0.14 0.003
AB 0.83 6.45* 5.81* 4.73 1.35 0.03 0.85 1.38 0.57
AC 1.23 3.28 8.88* 1.60 2.28 0.07 0.19 0.29 0.36
BC 2.20 0.001 2.85 6.66* 3.08 2.86 0.01 0.12
A2
B2 2.25 12.96 13.9*
C2 0.71
R2 0.53 0.86 0.74 0.87 0.71 0.29 0.30 0.20 0.53
ER expansion ratio, BD bulk density, TPC total phenolic content, AA antioxidant activity, HD hardness, CW
crispiness work, WSI water solubility index, WAI water absorption index, DD die diameter, Temp tem-
perature, SS screw speed
*indicates most significant terms at p \ 0.05

content (MC) of the product was found higher when they
were extruded through the die of 3 mm diameter. The
effect of barrel temperature and screw speed on moisture
content (MC) of produced extrudate from different die has
been presented in Fig. 1a, b. Speed of screw rotation was
found to have significant negative (p \ 0.05) effect on the
moisture retention in product for all barrel temperature, or
in other words, the extrudates produced at higher screw
speed contained lesser moisture for all barrel temperatures.
This might be caused due to the frictional heating of the
screw, which aids the heat content resulting in more
moisture evaporation. At high screw speed, the difference
between the moisture retention of products produced from
different die was reduced. Moreover, increase of barrel
temperature also caused a reduction in the moisture of
product in most of the experimental trials, which might be
due to enhanced moisture evaporation because of more
availability of heat at a higher temperature. The influence
of die diameter and barrel temperature and the interaction
terms was not statistically significant.
Expansion ratio
Expansion is one of the important characteristics of the
puffed product, which is controlled by the feed conditions
and processing variables. Expansion ratio (ER) of the
extrudates produced was found to lie in a range of 2.7–4.1.
The extrudate produced with the 3 mm die opening gained
higher puffing as compared to products from 3.5 mm as can
be observed from Fig. 1c, d. This might be because of the
more pressure generated inside the extruder in the case of
3 mm die opening, which facilitated a large pressure dif-
ference between inside and outside of the extruder. Table 1
showed that expansion of extrudates was observed to be
significantly influenced by the diameter of die, screw speed
and interaction of die diameter with barrel temperature
(p \ 0.05). Faster screw speed resulted in a significant
improvement in the expansion of the extruded product.
Increasing of screw speed causes higher absolute pressure
development inside the extruder. Therefore, the exit of
molten raw material from the die results into a sudden high-
pressure drop with the evaporation of more water, and con-
sequently, high expansion is achieved. At higher barrel
temperature (120 C), availability of more heat to convert
water into steam led to the greater expansion and effect of
increasing screw speed was diminished. The degree of
superheating of water in the extruder increases at a higher
temperature, leading to greater expansion in the product
improved expansion was also observed with increased screw
speed (p \ 0.05). The expansion takes place because of the
water vaporization, resulting from a sudden drop in the
pressure as the melted material exits from the die. Hence the
water flashes off, causing bubble formation in the extruded
material and making it porous (Sawant et al. 2013). Leonel
et al. (2009) also stated that under low moisture conditions,
an increase in barrel temperature led to greater expansion.
Bulk density
The bulk density of the puffed extrudates was varied
between 89 and 244.6 kg/m3. From the Table 1, it can be
clearly observed that density was significantly affected by
the screw speed of the extruder, interaction of die diameter
with temperature and screw speed (p \ 0.05). From the
Fig. 1e, f, it can be observed that increase in the temper-
ature from 110 to 120 C caused the bulk density to
decline, which could be attributed to the higher expansion
at a higher temperature for a product produced from 3 mm
die. Rising of barrel temperature results into increasing of
water superheating inside the extruder and causes higher

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Fig. 1 Effect of screw speed (a) 3 mm 100 °C (b) 3.5 mm 100 °C 110 °C 120 °C
and barrel temperature on a, 7 4.5

Moisture content (%db)

110 °C

Moisture content (%db)

b Moisture content; c, 6 4
d Expansion ratio; e, f Bulk 120 °C 3.5
5
density; g, h Water absorption 3
index (WAI); i, j Water 4 2.5
solubility index (WSI); k, 3 2
l Hardness; m, n Crispiness 1.5
2
work; o, p Total phenolic 1
content (TPC); q, r Antioxidant 1 0.5
activity (AOC) of the extrudates 0 0
250 300 350 250 300 350
Screw Speed (rpm) Screw Speed (rpm)

5 4
(c) 3 mm 100 °C 110 °C 120 °C (d) 3.5 mm 100 °C 110 °C 120 °C

4
3
Expansion rao

Expansion rao
3
2
2

1
1

0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

300 160
(e) 3 mm 100 °C (f) 3.5 mm 100 °C
140 110 °C
250 110 °C
Bulk density (kg/m3)

120 °C Bulk density (kg/m3) 120 120 °C

200
100
150 80
60
100
40
50
20
0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

12 11
(g) 3 mm 100 °C 110 °C 120 °C (h) 3.5 mm 100 °C 110 °C 120 °C
10 10.5

8 10
WAI (g/g)
WAI (g/g)

6 9.5

4 9

2 8.5

0 8
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

bubble formation along with a reduction in melt viscosity
which further contributes to declining of product density.
Moreover, speeding up of screw speed considerably
reduced the density because of high-pressure development
leading to greater expansion and lower density. In the case
of products from 3.5 mm diameter die, increase in tem-
perature alone didn’t affect the bulk density significantly,
however, simultaneous increase of temperature and screw
speed produced the product of lower bulk density. The
possible reason could be less residence time and high
sudden pressure drop creating highly expanded extrudates.
A similar trend of decrease in bulk density with tempera-
ture rise was also reported in many studies (Kaur et al.
2014; Ojokoh et al. 2015).

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Fig. 1 continued 30 100 °C 25

(i) 3 mm (j) 3.5 mm 100 °C 110 °C 120 °C
110 °C
25 20
120 °C
20
15

WSI(%)
WSI(%)
15
10
10
5
5

0 0
250 300 350 250 300 350

Screw speed (rpm) Screw speed (rpm)

4000 4500 (l) 3.5 mm

(k) 3 mm 100 °C 110 °C 120 °C 100 °C
3500 4000 110 °C
3000 3500 120 °C
Hardness(g)

Hardness(g)
3000
2500
2500
2000
2000
1500
1500
1000 1000
500 500
0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

4000 6000
(m) 3 mm 100 °C 110 °C
(n) 3.5 mm 100 °C
3500 120 °C
Crispiness work (g-mm)

110 °C
Crispiness work (g-mm)
5000
3000
4000 120 °C
2500

2000 3000
1500
2000
1000
1000
500

0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

250 180 100 °C

(o) 3 mm 100 °C
160
(p) 3.5 mm
110 °C
TPC (mg GAE/100 g)

TPC (mg GAE/100 g)

200 110 °C
140
120 °C 120 °C
120
150
100
80
100
60
50 40
20
0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

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 J Food Sci Technol

Fig. 1 continued 16 100 °C 110 °C 16

(r) 3.5 mm 100 °C 110 °C
(q) 3 mm
14 120 °C 14 120 °C

AOC (mg GAE/100 g)

AOC (mg GAE/100 g)

12 12
10 10
8 8
6 6
4 4
2 2
0 0
250 300 350 250 300 350
Screw speed (rpm) Screw speed (rpm)

Hydration properties (WAI and WSI)
Water absorption index (WAI): Extrudates from larger die
opening found to have higher WAI as can be observed
from Fig. 1g, h. From Table 1, it can be observed that only
diameter of the die was found to be a significant factor
affecting the WAI. The changes caused by other individual
and interaction terms were not significant. The WAI of
extrudates was varied from a minimum value of 6.1 to a
maximum value of 10.5 g/g. The products from 3 mm die
diameter showed decreased WAI on at higher temperature,
due to thermal breaking down of the starch into shorter
chain leading to decreased water holding capacity of the
molecules. However, increasing the screw speed led to
increasing the water absorption index at a barrel tempera-
ture of 100 and 110 C. As far products of 3.5 mm die
diameter are concerned, increasing speed (low shearing)
reduced the WAI was observed at a lower temperature, but
it was increased at a higher temperature because of the
higher damaged polymer chain and hydrophilic bonds at a
higher temperature and higher speed that reduced the
binding ability of molecules.
Water Solubility Index (WSI): Higher WSI of the pro-
duct was observed in the extrudates of 3 mm die diameter
as compared to that of 3.5 mm die diameter. Table 1
showed that die diameter, temperature and screw speed and
their interaction affected the WSI, but the influence was not
statistically significant. For the products made out of 3 mm
die opening, rise in screw speed and temperature resulted
in increased WSI. This indicated increased starch degra-
dation led by elevated temperature and increased screw
speed (Fig. 1i, j). Higher starch degradation resulted in
reduced entanglement and easily soluble molecules. Ojo-
koh et al. (2015) also highlighted a similar rise in WSI with
increased temperature in the extrudates of fermented maize
soybean blend. Lowest WSI was found at lower barrel
temperature with lower screw speed, and the highest value
of WSI was observed at the highest level of temperature
and screw speed as depicted in the figure. For extruded
products with 3.5 mm, WSI was increased with
temperature and seems constant with screw speed. Higher
WSI is often correlated with enhanced stickiness of
extruded products; hence, products with very high solu-
bility are less acceptable by consumers.
Hardness
Hardness and crispiness of the extrudates were not signif-
icantly affected by the independent variables and their
interaction terms (p [ 0.05) as shown in Tables 1. Increase
in the screw speed (Fig. 1k, l) caused a slight decrease in
the hardness (HD) at a lower temperature because of
lowered melt viscosity of the mix, which resulted in a
softer and less dense product. However, at the higher
temperature (120 C), result presented a small variation in
hardness of extrudate produced at different screw speed. In
the case of 3.5 mm products, at lower screw speed,
increase in barrel temperature showed a negative effect on
the hardness of the product. Further, it can also be observed
that higher temperature and increased screw speed impar-
ted an incremental effect on hardness. Moreover, the
hardness of the extrudates with 3 mm die opening was
found to be higher, as compared to the extrudate out of
3.5 mm die.
Crispiness work
Crispiness work (CW) of the products, extruded through
both the die was found to be reduced remarkably with the
rise in temperature for all screw speeds (Fig. 1m, n).
Combination of temperature and speed had a positive
interaction toward the decrement of crispiness. However,
according to some studies, crispiness increases with
increasing temperature (Brennan et al. 2011). Lowest
crispiness work was observed at maximum barrel temper-
ature and maximum screw speed, and on the other hand,
maximum CW was found at the lowest screw speed and
minimum temperature. Maximum crispiness work was
found in the product passed through die with 3.5 mm
diameter, as far as the die opening is concerned.

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J Food Sci Technol
Table 2 Predicted and
Name Goal
experimental values of
responses of fermented flour
extrudate under optimized Die diameter (mm) In range
process conditions
Temperature (C) Min
Screw speed (rpm) Min
Expansion ratio Max
Bulk density (g/cm3) In range
TPC (mg GAE/100 g) Max
AA (mg GAE/100 g) Max
WSI (%) In range
WAI (g/g) In range
ER expansion ratio, BD bulk density, TPC total phenolic content, AA antioxidant activity, HD hardness, CW
crispiness work, WSI water solubility index WAI water absorption index, DD die diameter, Temp tem-
perature, SS screw speed
Total phenolic content
Extrusion of the pre-fermented flour led to a sharp increase
in the total phenolic content (TPC) of the extrudate.
Extrusion processing enhances the total phenol content
from 63.47 mg GAE/100 g (fermented dough) to a maxi-
mum value of 210.3 mg GAE/100 g obtained at 100 C
temperature with screw speed 300 C and 3 mm die
diameter. In the case of 3.5 mm die diameter, the extrudate
produced under extrusion operation of 100 C temperature
with screw speed 350 C, presented a maximum value of
TPC content as 156 mg GAE/100 g. This significant
increase in the extruded product’s TPC is probably due to
liberation of bound phenolic compounds resulting from the
cell wall and cellular constituent’s breakdown. Greater
liberation of the phenolic compounds is obtained from the
extrudates with 3 mm die diameter probably due to higher
pressure. It can be observed from the Fig. 1o, p that
increasing of barrel temperature led to a significant
reduction (p \ 0.05) in TPC, which might be due to dis-
integration of phenolic compounds or change in the
chemical reactivity, reduced extractability because of a
certain degree of polymerization. The figure also revealed
that increased TPC at higher Screw speed was found in the
extrudates from 3.5 mm die diameter. This is probably due
to high shearing and tissue disintegration action. Table 1
revealed that along with barrel temperature, a combination
of barrel temperature and screw speed was also found to be
a significant term affecting the TPC content of extrudates.
Simultaneous increase of temperature and screw speed
shows higher negative effect on the phenolic content than
individual effect. Research carried out by Rathod and
Annapure (2017) also revealed that the extrusion process of
lentil-orange peel powder increased the TPC and Antioxi-
dant content by 70.4% and 60.6%, respectively. Altan et al.
(2009) also found a remarkable reduction in TPC content at
the higher temperature (140 C, 150 C) in the extrusion
Importance Predicted values (PV) Experimental values (EV)
(D = 0.85)
3 3 3
4 100 100
3 289 289
4 3.37 3.476
4 0.167 0.111
4 159.86 161.43
4 12.93 9.8
3 16.61 18
3 8.66 8.55
cooking of barley-fruit product at a screw speed of
150 rpm and 200 rpm.
Antioxidant activity
Effect of extrusion on AA was similar to the effect of
extrusion on TPC; however, the results were not signifi-
cantly affected by the independent variables as can be
observed from Table 1. Antioxidant activity of the fer-
mented dried flour was 7.41 mg GAE/100 g, which
increased up to a maximum value of 13.89 mg GAE/100 g
in product extruded from 3.5 mm die at 120 C and
250 rpm. The extrudates produced from 3 mm die with a
barrel temperature of 100 C and a screw speed of 250 rpm
showed an antioxidant content of 13.10 mg GAE/100 g.
Hence, findings indicated an improvement in antioxidant
activity upon extrusion. However, increasing of barrel
temperature and screw speed caused a decline in antioxi-
dant activity as observed in Fig. 1q, r and presented a
noticeable reduction with simultaneous increase of screw
speed and temperature as compared to their individual
effect. Increased antioxidant activity was observed with
increased screw speed at lower barrel temperature, for the
products made out of 3.5 mm die diameter. This variation
might be caused due to greater mechanical energy input,
which led to heat generation and subsequent degradation of
antioxidant elements. The decline in antioxidant activity
was also reported in various food materials, as processing
temperature increases (Altan et al. 2009; Charunuch et al.
2014).
Principal component analysis and clustering
The principal component analysis of variables indicated
that observations of different variables were found to be
relatively spherical as three PCs were observed to be sig-
nificant with a cumulative explained variance of about
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 J Food Sci Technol

Fig. 2 Principal component

analysis, a Loadings plot of
different variables;
b Dendrogram plot for cluster
analysis; Sample Notation:
sample no. Sijk stands for
sample produced at ith level of
temperature (100, 110, 120 C),
jth level of Screw speed (250,
300, 350 rpm) and kth level of
die diameter (3 & 3.5 mm)

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J Food Sci Technol
Table 3 Combined effect of the fermentation and extrusion on physicochemical properties of rice-black gram flour extrudates
S. No Variable Unfermented flour
1 ER
2 BD (g/cm3)
3 WSI (%) 6.8
4 WAI (g/g) 2.95
5 TPC (mg GAE/100 g) 35.625
6 AA (mg GAE/100 g) 6.53
ER expansion ratio, BD bulk density, TPC total phenolic content, AA antioxidant activity, WSI water solubility index, WAI water absorption
index
86%. From the principal component analysis, it was
observed that only 32.57% of the total variance in the
samples was explained by 1st principal component (PC1)
and 30% by 2nd principal component (PC2), however,
studied variables were significantly correlated (Bartlett test,
p \ 0.0001). Loading plot shown in Fig. 2a, revealed that
TPC–AOC and BD–MC were highly positively correlated,
while WSI–WAI and MC–ER were negatively correlated.
The moisture content in the extrudates increases its bulk
density because water will be entrapped within the inter-
cellular voids in the sample. Higher expansion leads to an
increase of volume due to more number of air bubbles
formed that results in lowering of the bulk density. Since
major phenolic compounds show antioxidant activity hence
TPC and AOC were found to be positively correlated.
Negative correlation existing between WSI and WAI
indicates the increase of solubility index with a reduction
of water absorption capacity of the extrudates. In addition
to it, TPC, AOC and WAI were found to be uncorrelated
with BD, ER or MC, whereas, WSI has a correlation with
ER as well as TPC and AOC. Further, to classify all the
samples based on their physicochemical properties, as PC1
and PC2 alone can’t be used for a confirm interpretation,
hierarchical clustering of samples was made using three
principal components all together which explained about
85.6% variation. A dendrogram plot with euclidean dis-
tance-centroid linkage method was drawn, as shown in
Fig. 2b. Considering the sample S111 and S231 outliers, all
the samples can be grouped into two classes; one for 3 mm
dia and another for 3.5 mm diameter die as represented in
the dendrogram. Thus, it can be inferred that die diameter
has a potential effect on the physicochemical properties of
extrudates.
Optimization and validation of the extrusion
conditions
The quality of the puffed extruded product depends upon
the raw materials composition as well as extruder operating
conditions. Hence numerical optimization of process
Fermented flour Unfermented extrudate Fermented flour extrudate
3.662 3.476
0.1081 0.1117
9 15.76 18
2.84 9.172 8.545
63.47 52.57 161.43
7.41 9.19 9.8
variables was done to achieve desired physical properties
and improved functional properties of extruded products
(Singh et al. 2015). Since lower temperature caused
remarkable extraction of TPC and antioxidant activity. So
minimum temperature and screw speed were targeted in the
optimization process. The constraints used for the opti-
mization of the process, along with optimized and pre-
dicted data had been reported in Table 2. The verification
experiments were done under the optimum extrusion con-
ditions to validate the results predicted by software.
Comparison between fermented flour extrudates
and unfermented flour extrudates properties
The produced extrudates using fermented, and unfermented
flour as raw material at optimized conditions (100 C BT,
289 rpm SS and 3 mm die opening) were analyzed for
physicochemical properties. Expansion, bulk density, WSI
and WAI of both extrudates differed with a slight differ-
ence (Table 3). The remarkable observation was higher
TPC and AA obtained from fermented flour extrudate than
unfermented flour extrudate. This is resulted because of the
fermentation process. Fermentation also improved water
solubility, however reduced the WAI. Reduction in bulk
density and increase in WAI was stated by Ojokoh et al.
(2015) in the fermented maize soya bean extrudates when
compared to unfermented blend extrudates. Higher WSI in
the fermented flour represents the accelerated degradation
of the starch molecules during extrusion. Hence increased
WSI and decreased WAI is the combined effect of the
fermentation and extrusion. From the results, it was also
cleared that extrusion also improves the extraction of the
bio-actives compounds.
Sensory evaluation of the puffed extrudates
The sensory scores and importance to quality parameters
given by the panel were used for calculation of similarity
values using the fuzzy logic approach. The unfermented
flour extrudates and fermented flour extrudates have been
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 J Food Sci Technol

named as S1and S2 respectively. The brief of stepwise judges’ preference to importance of quality attributes were
results obtained from the fuzzy analysis of sensory scores calculated.
of both types of products has been shown below. QC ¼ ð48:43; 25; 25Þ

Triplets associated with the sensory scales QTs ¼ ð76:56; 25; 18:75Þ
QTx ¼ ð82:81; 25; 12:5Þ
Triplets is a set of three numbers, representing triangular
QM ¼ ð65:62; 25; 23:43Þ
membership function. For a 5-point hedonic scale, triplets
are as follows: poor/not at all important (0, 0, 25); Fair/ QA ¼ ð67:18; 25; 20:31Þ
Somewhat important (25, 25, 25); Good/Important (50, 25,
25); very good/ highly important (75, 25, 25); excellent/
Triplets for Relative weightage of quality attributes
extremely important (100, 25, 0).
For the calculation of the triplets for relative weightage,
Triplets for sensory score of extruded samples
sum (Qsum) of first digits of the triplets of QC, QTs, QTx, QM,
QA was obtained.
For the colour of sample 1 (S1C), triplets are calculated
using the following Eq. 9.

½0ð0; 0; 25Þ þ 0ð25; 25; 25Þ þ 5ð50; 25; 25Þ þ 9ð75; 25; 25Þ þ 2ð100; 25; 0Þ
S1C ¼ ð9Þ
0þ0þ5þ9þ2

In the similar way, the triplets for all the quality attri- Qsum ¼ ð48:43 þ 76:56 þ 82:81 þ 65:62 þ 67:18Þ
butes for both the samples were determined, which has ¼ 340:625 ð10Þ
been listed as follow:
Triplets for relative weightage for colour:
S1C ¼ ð70:31; 25; 21:87Þ
S1c
S1Ts ¼ ð68:75; 25; 21:87Þ QCrel ¼ ð11Þ
Qsum
S1Tx ¼ ð68:75; 25; 20:375Þ QCrel ¼ ð0:1421; 0:0733; 0:0733Þ
S1M ¼ ð73:43; 25; 20:31Þ QTsrel ¼ ð0:2247; 0:0733; 0:0550Þ
S1A ¼ ð68:75; 25; 21:87Þ QTxrel ¼ ð0:2431; 0:0733; 0:0366Þ
S2C ¼ ð71:87; 25; 20:31Þ QMrel ¼ ð0:1926; 0:0733; 0:0688Þ
S2Ts ¼ ð78:125; 25; 18:75Þ QArel ¼ ð0:1972; 0:0733; 0:0596Þ
S2Tx ¼ ð81:25; 25; 15:625Þ
S2M ¼ ð81:25; 25; 12:5Þ Table 4 Similarity values of extrudates from unfermented flour (S1)
and fermented flour (S2) obtained from fuzzy logic analysis of sen-
S2A ¼ ð73:43; 25; 15:625Þ sory data
S1 denotes the sample 1 (extrudates from unfermented Scale S1 S2
flour) and S2 denotes the sample 2 (extrudates from fer-
Poor (F1) 0.000 0
mented flour); C: Colour; Ts: Taste; Tx: Texture; M:
Fair (F2) 0.11248 0.062
Mouthfeel; A: Aftertaste.
Satisfactory (F3) 0.3956 0.28609
Triplets for judges’ preference to importance of quality Good (F4) 0.7200 0.655
attributes Very good (F5) 0.65038 0.668
Excellent (F6) 0.230 0.284
Using the similar method as followed in ‘‘Triplets for Bold represents the maximum similarities values of samples
sensory score of extruded samples’’ section, the triplets for S1, unfermented flour extrudate; S2, fermented flour extrudate;
F1,F2,…F6 represents membership function for similarity values

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J Food Sci Technol

Fig. 3 Scanning electron microscopy images of a Unfermented rice- 5000 X, respectively; e, f Fermented flour extrudate at magnifications
black gram flour at 5000 X; b Fermented rice-black gram flour at at 600 X and 4500 X, respectively
5000 X; c, d Unfermented flour extrudate at magnifications 650 X and

123

Triplets for overall sensory score of extrudates
For an overall sensory score of extrudates, triplets (SO1)
and (SO2) were estimated using the following equation.
The following rule (Eq. 12) was adopted for the multipli-
cation of the triplets:
ða; b; cÞ  ðd; e; f Þ ¼ ða  d; a  e þ d  b; a  f þ d  cÞ
ð12Þ
SO1 ¼ S1c  QCrel þ S1Ts  QTsrel þ S1Tx  QTxrel þ S1M
 QMrel þ S1A  QArel
ð13Þ
SO1 ¼ ð69:8735; 50:6888; 41:8168Þ
SO2 ¼ ð77:6710; 53:3264; 38:9207Þ
Values of membership function of standard fuzzy logic
scale
A set of 10 values defines values of the membership
function of F1–F6.
F1 ¼ ð1; 0:5; 0; 0; 0; 0; 0; 0; 0; 0Þ
F2 ¼ ð0:5; 1; 1; 0:5; 0; 0; 0; 0; 0; 0Þ
F3 ¼ ð0; 0; 0:5; 1; 1; 0:5; 0; 0; 0; 0Þ
F4 ¼ ð0; 0; 0; 0; 0:5; 1; 1; 0:5; 0; 0Þ
F5 ¼ ð0; 0; 0; 0; 0; 0; 0:5; 1; 1; 0:5Þ
F6 ¼ ð0; 0; 0; 0; 0; 0; 0; 0; 0:5; 1Þ
Values of overall membership function of sensory scores
on the standard fuzzy scale
For a given value of x on the abscissa, the value of
membership function Bx is expressed as shown by Eq. 14.
½ x  ð a  bÞ 
Bx ¼ for ða  bÞ\x\a
b ferent from that of raw material as illustrated in Fig. 3a, b.
½ða þ cÞ  x ð14Þ
¼ for a\x\ða þ cÞ
c processing showed undamaged starch granules along with
¼ 0 for all other values of x
x ¼ ð0; 10; 20; 30; 40; 50; 60; 70; 80; 90; 100Þ
B1 ¼ ð0; 0:159; 0:21; 0:41; 0:60; 0:80; 1; 0:99; 0:75; 0:51Þ
B2 ¼ ð0; 0; 0:105; 0:293; 0:481; 0:668; 0:856; 1; 0:94; 0:683Þ
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J Food Sci Technol
Similarity values for extruded samples
Membership function for both the extrudates are defined by
B1, B2 provided in the Sect. 3.4.8. The similarity values
(Sm) for the samples were calculated by the Eq. 15.
SmðF; BÞ ¼ FoB0 =maximum of ðFoF; BoB0 Þ ð15Þ
0
FoB = product of matric F with the transpose of matric
B.
FoF = product of matric F with the transpose of matric
F.
BoB0 = product of matric B with its transpose.
The calculated similarity values of both the samples
have been listed in Table 4. The maximum similarity value
of a sample defines the quality of that sample. From the
table, the maximum similarity values for the sample 1 was
0.72 (under good scale factor), and for the sample 2 was
0.668 (under very good scale factor). This showed that the
product from the fermented flour (sample 2) had higher
overall acceptability than unfermented flour extrudates.
Scanning electron microscopy analysis
Micrographs of the raw materials, i.e., fermented and
unfermented rice-black gram flour and extruded products
developed at optimized extrusion conditions are shown in
Fig. 3. It can be observed from Fig. 3a, that unfermented
sample’s particles were strongly embedded, compact and
presents a smooth morphology at a magnification of 5000
X. Fermented sample showed a presence of irregular, rough
and porous morphology as can be clearly visualized from
Fig. 3b. At a magnification of 600–650 X, SEM images
showed the porous open-celled and sheared structures as
depicted in Fig. 3c–e. Porous internal structures of extru-
dates prepared from unfermented and fermented flour is
due to the formation of air voids during the transformation
of water into steam. Micrographs obtained at a magnifi-
cation of 4500–5000 X (Fig. 3d–f), represented the com-
pact embedded surfaces of the gelatinized starch.
Morphology of the extruded product was very much dif-
Micrographs of raw material obtained before the extrusion
pores and cavities. However, after the extrusion cooking,
the microstructure of extrudates at 5000 X magnification
revealed a continuous and compact structure with the lack
of starch granules. This indicates the complete melting of
starch because of exposure to high temperature (110 C)
along with severe shearing and rupturing due to high screw
speed (300 rpm) during extrusion cooking. Although, as
such, no much difference is observed between fermented
and unfermented extrudates at same extrusion conditions.
J Food Sci Technol
Conclusion
In this study, fermented rice-black gram flour was used for
the development of ready to eat puffed snack. It was found
that the combination of the fermentation and extrusion
process improved the extractability of the bioactive com-
pound. Increase in the barrel temperature effects the
bioactive compounds negatively, and higher screw speed
helps in better extractability of phenolic compounds.
Principal component analysis loading plot revealed that
TPC–AOC and BD–MC were highly positively correlated,
while WSI–WAI and MC–ER were found to be negatively
correlated. Extrusion of the rice-black gram flour led to
complete melting of the starch as represented by the
compact and continuous microstructure of the extruded
products. The optimum conditions for the development of
the puffed product from 10% moisture fermented rice black
gram flour as obtained through numerical optimization
were found to be 100 C barrel temperature, 289 rpm
screw speed and 3 mm die opening. Moreover, higher
similarity value of extrudates of fermented flour over
unfermented flour extrudates, obtained from the fuzzy logic
approach indicated higher overall acceptability of fer-
mented flour-based snacks. Hence, present research
demonstrated that the potential of fermented rice-black
gram flour for development of nutritionally rich and heal-
thy ready to eat snack product through extrusion
processing.
Acknowledgements The authors also like to acknowledge the
assistance provided by Mr Nirmal Dolai for experiments and extruder
operation during the course of this work.
Compliance with ethical standards
Conflict of interest There are no conflicts of interest to report.
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