J Food Sci Technol
https://doi.org/10.1007/s13197-019-03916-1
ORIGINAL ARTICLE
Optimal fermentation parameters for processing high quality
African locust bean condiments
Bidossessi Pélagie Agbobatinkpo1 • Gbètognon Marc Tossou1 • Laurent Adinsi1
Houédougbé Noèl Akissoe1 • Djidjoho Joseph Hounhouigan1
Revised: 19 February 2019 / Accepted: 2 July 2019
 Association of Food Scientists & Technologists (India) 2019
Abstract Fermented African locust bean (ALB) condi-
ments are widely used to enhance the aroma and the taste
of various dishes in West Africa. They are a good source of
proteins. Sonru is one of the ALB condiments processed in
Benin using a traditional starter called yanyanku. The
variability in sonru processing unit operations (e.g. fer-
mentation) presumably affects the quality (microbiological,
biophysical and sensorial) of the final product. This study
aimed at optimizing the fermentation step of sonru based
on response surface methodology including three-factors
[i.e. proportion of yanyanku, fermentation duration and
thickness/depth of the layer of ALB (TALB)] and three-
level factorial Box Behnken design with four central point
replicates. The pH, softness of fermented cotyledons,
Bacillus spp. and aerobic mesophilic bacteria count of
sonru were evaluated. In addition, consumer acceptability
was assessed by 7-hedonic box scale and Just About Right
analysis. The pH had an upward trend with the increasing
of all factors, and particularly Bacillus spp. count had
similar trend with the fermentation duration and TALB
whereas maximal penetration force (referred as the soft-
ness) had a downward trend with the fermentation duration.
The optimal fermented ALB condiment resulted from
processing conditions set at 1% of yanyanku, 24 h of fer-
mentation and 8.5 cm for TALB. The optimized sonru
appeared to be ideal in color, softness and taste. These
findings can be used to improve fermented ALB condi-
ments processing and better control the product quality in
& Bidossessi Pélagie Agbobatinkpo
vodabpelagie@gmail.com; abpelagie@yahoo.fr
1
Faculté des Sciences Agronomiques, Université d’Abomey-
Calavi, 03 BP 2819 Jericho, Cotonou, Benin
•
African countries. Further studies are needed to improve
the odor of sonru.
Keywords African locust bean  Sonru  Process
optimization  Consumer acceptability  Penalty analysis
Introduction
Traditional fermented foods are an important part of the
diet in Africa (El Sheikha and Montet 2014). The fer-
mented condiments obtained from ALB (Parkia biglobosa)
are largely used in the diet of people in West Africa (Ouoba
et al. 2008). They are known as soumbala in Burkina Faso
(Ouoba et al. 2003), dawadawa in Ghana and Nigeria
(Parkouda et al. 2009; El Sheikha and Montet 2014) and
afitin, iru or sonru in Benin (Azokpota et al. 2006). They
are used to enhance the flavor of many dishes including
soup and sauces because of their sensory characteristics
and high nutritional value (Azokpota et al. 2006; Parkouda
et al. 2009). They are significant low-cost sources of pro-
tein and lipid with high levels of free amino acids, such as
cysteine, methionine, leucine, isoleucine, tyrosine, pheny-
lalanine, and even lysine, which is limiting in plant foods
(Agbobatinkpo et al. 2011). The fermentation of ALB is
spontaneous and generally there are no national standards
for the product in West African countries, leading to
variations in stability, nutritional and hygienic quality of
the final product (Azokpota et al. 2006; Parkouda et al.
2009). The fermentation duration depends on the local
conditions and the consumer demand/specification (Odunfa
1988; Azokpota et al. 2006). The success of the fermen-
tation indeed depends on several parameters including the
development of Bacillus spp. which is closely affected by
the temperature of food matrix. Equipment with specific
123

surface characteristics such as semi-porous clay, charred
wood, gourds, etc., is one of a critical challenge to the
improvement of some traditional fermentation processes
(El Sheikha 2018). In Benin, the fermentation is conducted
in wooden tray where the dehulled ALB are laid down in
layer of approx. 2 cm allowing a temperature of around
30–36 C inside the product during the fermentation. A
high thickness of the layer of ALB cotyledons in the fer-
mentation tray could allow optimal growth temperature for
Bacillus spp. which are the main microorganisms involved
in ALB fermentation (Ouoba et al. 2004); consequently
improve safety of the fermented product.
To improve the fermentation process of indigenous fer-
mented products, traditional starters were developed.
Example of these starters are yanyanku and iku-iru (Azok-
pota et al. 2006), ikpiru (Agbobatinkpo et al. 2011) used for
ALB fermentation, daqu used for alcoholic fermentation
(Zheng et al. 2011); loog-pang for traditional drink (Kishida
et al. 2009). Yanyanku is a fermented product based on
Hibiscus sabdariffa seeds used as traditional starter to pro-
duce sonru, an ALB fermented condiments widely con-
sumed in Benin (Azokpota et al. 2006). Yanyanku played a
role at softening the dehulled ALB during fermentation
(Agbobatinkpo et al. 2012) and the rate of softening depends
on the quality of yanyanku and the ratio yanyanku/dehulled
ALB (Agbobatinkpo et al. 2011). Indeed, the softening of
ALB cotyledons is one of the quality criterion used tradi-
tionally to check the success of fermentation. Agbobatinkpo
et al. (2011) reported variability in the proportion of
yanyanku used by sonru processors. El Sheikha (2018)
mentioned that only when conditions such as time, temper-
ature, pH, substrate pretreatment, and inoculum-substrate
ratio are controlled that fermentation can be optimized. This
study aims to evaluate the effect of the ratio yanyanku/ALB
seeds, the fermentation duration and the thickness/depth of
the layer of ALB cotyledons in the fermentation tray through
Response Surface Methodology (RSM) design on selected
quality parameters of sonru to define the optimal conditions
of ALB dehulled beans processing to produce sonru suit-
able for consumers acceptance.
Fig. 1 a Wooden tray for sonru
fermentation; b plain sonru a b
10.5cm
4 cm
123
J Food Sci Technol
Materials and methods
Material
Malcavene beans (H. sabdariffa) and ALB (P. biglobosa)
were used to produce yanyanku and sonru, respectively.
Seeds of H. sabdariffa and P. biglobosa were bought from
a farmer at Parakou (North of Benin) during August 2017.
The fermentation of ALB was conducted in wooden tray
(length: 40 cm, wide: 25 cm and height varying from 4 to
17 cm) (Fig. 1a).
Yanyanku and sonru production
Yanyanku (traditional starter) was produced by cooking
malcavene bean in ash filtrate solution as described by
Agbobatinkpo et al. (2011). For sonru preparation, raw
African locust beans (ALB) were processed according to
the traditional method reported by Azokpota et al. (2006).
To summarize, ALB seeds were boiled for 12 h and left
overnight in the boiling water. The testa of ALB was
removed by fulling and the beans were washed and cooked
again for 1 h (Fig. 2). Yanyanku was added to the ALB
cotyledons 15 min before the end of the second cooking.
Cotyledons were spread into close wooden tray for fer-
mentation. The proportion of yanyanku (PY), the fermen-
tation duration (FD) and the thickness of the layer of ALB
cotyledons in tray fermentation (TALB) varied according
to the experimentation (Table 1).
Experimental design
A Box–Behnken response surface methodology (RSM)
design as described by Myers et al. (2009) and Ying et al.
(2014) was used to determine the optimal conditions for
ALB fermentation during sonru processing. Therefore,
three factors including proportion of yanyanku (8.4–120 g
for 100 kg of ALB cotyledons), fermentation duration
(6–30 h) and thickness of the layer of ALB cotyledons in
fermentation tray (2–15 cm) were experimented on the
17cm
J Food Sci Technol

Dried African locust beans

Sorting and washing

First cooking 12 h

Dried Hibiscus
Cooling About 30 min
sabdariffa seeds

Fine sand Dehulling

Washing
Washing mud
Ash filtrate solution: Sorting in water
(ash/water, ½) 330 ± 117 Cooking (1 h) + testa
mL/kg dried seeds)
Cotyledons + whole beans
Draining
Mud soil
Fermentation (30 h) Sorting in water Whole beans
solution
Drying Dehulled beans

Pounding
Washing

Yanyanku Seconde cooking 1 h

Cooked cotyledons

Fermentation 30 h

Sonru

Fig. 2 Flow sheet of sonru processing. Source: Azokpota et al. (2006) and Agbobatinkpo et al. (2011)

selected quality parameters of sonru, i.e. pH, texture
(softness indices) of fermented cotyledons, Bacillus spp.
and aerobic mesophilic bacteria count. The levels of factors
were defined based on previous studies (Azokpota et al.
2006; Agbobatinkpo et al. 2012) with increase of thickness
of the layer of ALB cotyledons in order to give better
condition for the growth of Bacillus spp. being the main
microorganisms for ALB fermentation. The Box–Behnken
RSM design provided 16 trials (Table 1) with four central
point replicates were conducted in laboratory scale for
sonru production.
Microbiological and biophysical analyses
The count of Aerobic mesophilic bacteria (AMB) was
assessed on Plate Count Agar medium (Oxoid, CM 463,
Basingstoke, Hampshire, England) after incubation for
72 h at 30 C. For Bacillus spp. count, 5 mL of inoculum
was heated at 80 C for 10 min, serially diluted and enu-
merated on Nutrient Agar (Oxoid, CM 003, Basingstoke,
Hampshire, England) after incubation for 48 h at 37 C.
The pH was measured with a pH meter (Inolab 730)
according to Nout et al. (1989). The maximal penetration
forces (referred as softness indices) of fermented cotyle-
dons were determined with uniaxial penetration test which
were performed on fermented cotyledons samples using a
Stevens texture analyzer (Stevens-LFRA texture analyzer,
Harlow, UK). Each cotyledon was compressed with a
conical probe (Clear Plastic 28 mm diameter TA16 for
penetration) moving at 0.2 mm/s over total displacement of
2 mm.

123
 J Food Sci Technol

Table 1 Variable combinations in experimental runs and effect of independent variables on penetration force, pH and AMB and Bacillus spp.
counts of sonru
Experiments Variables Reponses
Fermentation TALB Proportion of Yanyanku Texture pH AMB count Bacillus spp. count
duration: (FD) (h) (cm) (10-3)/g ALB (penetration force) (log CFU/g) (log CFU/g)
(N)

1 6 2 0.64 2.47 6.4 6.4 7.5

2 30 2 0.64 0.08 8.4 9.8 10
3 6 15 0.64 2.31 6.4 8.1 8.8
4 30 15 0.64 0.45 6.5 9.7 10.3
5 6 8.5 0.08 1.33 6.4 7.8 8.1
6 6 8.5 1.2 2.22 6.8 8.2 8.8
7 30 8.5 0.08 0.3 7.2 11.2 12.1
8 30 8.5 1.2 0.09 7.9 11 11.4
9 18 2 0.08 0.23 6.9 10.1 9.2
10 18 15 0.08 0.43 6.7 10.4 10
11 18 2 1.2 0.18 7.6 10.3 10.6
12 18 15 1.2 0.63 6.8 9.9 11
13 18 8.5 0.64 0.41 6.9 10.8 11.1
14 18 8.5 0.64 0.41 6.9 11 10.1
15 18 8.5 0.64 0.41 6.8 9.3 10.5
16 18 8.5 0.64 0.24 6.9 8.9 10.8
SD# 0.06 0.03 0.9 0.32
: central point, SD# standard deviation of four replicates central point

Ethical assessment and consent
A brief description of the sensory tests was made to con-
sumers in order to collect their notice of consent. Only
those who have given their agreement participated in these
sensory tests and they are free to withdraw at any time.
Consumer acceptability
A ranking test was performed to complete the softness
indices (texture) of fermented cotyledons. Ten (10) of
sixteen (16) experimental samples were randomly selected.
Ten (10) street food sellers who frequently used sonru in
food preparation ranked the softness of ten samples on
scale from 1 to 10 by using toothpick.
Furthermore, the sonru produced from optimal condition
was evaluated by 76 consumers in two forms: plain sonru
(Fig. 1b) and sonru incorporated into cooked rice. The
plain sonru and rice (ratio sonru/rice: 6/10) were cooked
together with tap water without salt.
The evaluation was carried out through a 3-point Just-
About-Right (JAR) scale to determine intensity of attri-
butes (i.e. too weak, just-about-right, too strong) that can
affect the overall acceptance of sonru. The descriptors
color (color characteristic of sonru), odor (odor
characteristic of sonru) and softness (means fermented
cotyledons softening: if it is too or less soften or good
for consumers) were evaluated on plain sonru whereas
taste (taste known for sonru) and flavor (flavor charac-
teristic of sonru perceptible after tasting) were evaluated
on cooked sonru with rice. After JAR test, the consumers
evaluated the overall acceptability of sonru on a 7-point
hedonic scale (1 = dislike very much and 7 = like very
much).
After testing the products, the consumers were inter-
viewed for gathering information on gender, age, occupa-
tion and frequency of consumption.
Statistical analysis
The experimental data collected from Box–Behnken
design were analyzed by the response surface regression
procedure using second-order polynomial equation. All
these statistical analyses and the graphical presentations
were performed using Statistica software (version 7.1,
Stat Soft France 2006). The data collected from overall
liking were analyzed for the consumer acceptability while
those from JAR test were analyzed by penalty analysis
(XLstat—pro, Addinsoft 2012) to determine the ideal
quality product.

123
J Food Sci Technol

Results and discussion
Effect of fermentation duration (FD), thickness/
depth of the layer of ALB cotyledons (TALB)
and proportion of yanyanku (PY)
The responses on dependent variables (Bacillus spp. count,
AMB count, pH and texture of fermented cotyledons) were
tested in the three dimensions model. The coefficient of
determination (R2) is defined as the proportion of variation
in the response attributed to the model rather than to ran-
dom error, with R2 being higher than 80% for a good fitted
model (Krishnaiah et al. 2011). In this study, the models
produced high coefficients of determination (86–99%) for
the Bacillus spp. count, the AMB count, the pH and the
texture of fermented cotyledons indicating that the
regression models explained very well the experimental
design (Table 2).
Aerobic mesophilic bacteria (AMB) count
The model explained 86% of the variations in AMB count,
with related formula describing the behavior of the AMB.
AMB ¼ 3:8 þ 4  102 X1 7  103 X21 þ 3
 102 X2 102 X22 5  101 X3 þ 8  101 X23
5  103 X1 X2 2  102 X1 X3 5  102 X2 X3
where X1 (fermentation duration: FD), X2 (thickness of the
layer of ALB cotyledons: TALB) and X3 (proportion of
yanyanku: PY) = independent variables.
The fermentation duration (FD) was the only factor that
significantly influenced the AMB count. This factor had
positive value for both linear (p = 0.003) and quadratic
(p = 0.037) effect (Table 2). The absence of the proportion
of yanyanku (PY) effect on AMB count was not expected
because the yanyanku was used with the aim to increase the
microorganisms’ development. However, previous works
(Agbobatinkpo et al. 2012) that compared the sonru pro-
duction with yanyanku and without yanyanku showed
significant effect of yanyanku on AMB count. Thus, the use
of yanyanku had significantly influenced the development
of AMB count in sonru but the increase of yanyanku
proportion has no effect as long as it is greater than 0.6 g
for 100 kg of ALB. Likewise, the results showed no effect
of thickness/depth of the layer of ALB cotyledons (TALB)
on AMB count. In this study, the thickness of the layer of
ALB cotyledons in tray fermentation varied from 2 to
15 cm allowing different profiles of temperature. However,
the increase of temperature did not show any effect on
AMB count. During the first 12 h of fermentation, the
temperature of the batch varied from 30 to 36 C and from
30 to 47.5 C for 2 and 8.5 cm of thickness of the layer of
ALB cotyledons in fermentation tray respectively. No
significant interaction was observed between independent
variables (Table 2). A response surface plot (Fig. 3a)
showed similar AMB count whatever the PY, for the same
FD. The maximum count of AMB was reached around
16–18 h of fermentation. This result is consistent with
Agbobatinkpo et al. (2012) who reported that during ALB
fermentation, AMB reached maximun count (10.08 Log10
UFC/g) at 18 h of fermentation.
Bacillus spp. count
The fitted model equation obtained for Bacillus spp. count,
with high determination coefficient (R2 = 91%), indicated
the adequacy and good precision of the statistical model:

Table 2 Coefficient of
Bacillus spp. count AMB count pH Texture (penetration force)
determination (R2) and
probabilities indicating the R2 0.91 0.86 0.97 0.99
effects of factors on dependent
variables of sonru Fermentation duration (FD)
LE 0.003(?) 0.003(?) \0.001(?) \0.001(-)
(?) (?) (-)
QE 0.037 0.037 0.502 \0.001(-)
Proportion of Yanyanku/g ALB (PY)
LE 0.225(?) 0.966(-) 0.03(?) 0.183(?)
(-) (-) (-)
QE 0.433 0.196 0.133 0.131(?)
TALB (EB)
LE 0.164(?) 0.544(?) \0.001(-) 0.167(?)
QE 0.06(?) 0.304(?) 0.896(?) 0.125(-)
FD 9 TALB 0.427(-) 0.311(-) \0.001(-) 0.215(?)
(-) (-) (?)
FD 9 PY 0.281 0.727 0.464 0.029(-)
(-) (-) (-)
TALB 9 PY 0.824 0.642 0.140 0.557(?)
Probabilities in bold indicated significant effect
LE linear effect, QE quadratic effect, (?)positive effect of factor on dependent variable, (-)negative effect of
factor on dependent variable

123
 J Food Sci Technol

a Bacillus spp (log cfu/g) b Aerobic mesophilic bacteria (log cfu/g)

16 1,4

14
1,2

12

Ratio yanyanku / ALB (10-3)

1,0

10
TALB (cm)

0,8
8

0,6
6

4 0,4

2
11 0,2
10 11
9 10
0
8 9
4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 0,0
7 8
Fermentation duration (h) 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32
7
Fermentation duration (h)

c pH d Texture: Penetration force (N)

1,4 1,4

1,2 1,2
Ratio yanyanku / ALB (10-3)

[Ratio yanyanku/ALB] (10-3)

1,0 1,0

0,8 0,8

0,6 0,6

0,4 0,4

0,2 0,2
2,5
8 2
7,6 1,5
7,2 1
0,0 6,8 0,0 0,5
4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32
6,4 0
Fermentation duration (h) Fermentation duration (h)

Fig. 3 Surface plots showing the effect of dependents variables on pH, penetration force and Bacillus spp. and AMB count of sonru

Bacillus spp: count ¼ 4:04 þ 3  101 X1 5  103 X21 by considering that Bacillus spp. were in spore forms
1 2 during the first 12 h of ALB fermentation (Agbobatinkpo
þ 4  10 X2 10 X22 þ 5
et al. 2012) and of others beans fermentation such as
 101 X3 þ 8  101 X23 roselle beans (Parkouda et al. 2009). Indeed, Bacillus spp.
3  103 X1 X2 5  102 X1 spores require transformation into vegetative state and the
duration of endospore transformation into vegetative state
X3 102 X2 X3
is environmental condition, specie and strain dependent
ðX1 ¼ FD; X2 ¼ TALB and X3 ¼ PYÞ (Sneath 1986; Agbobatinkpo et al. 2012). Bacillus spp.
reached maximum count (11 Log10 cfu/g) at 14 h of fer-
Fermentation duration (FD) and thickness/depth of the mentation when the TALB was beyond 6 cm versus at 16 h
layer of ALB cotyledons in fermentation tray (TALB) had of fermentation when the TALB was below 6 cm but this
significantly and positively quadratic effect on Bacillus interaction was not significant (p = 0.427).
spp. count. In addition, FD had also a positive linear effect The proportion of yanyanku had no significant effect on
(p \ 0.005) (Table 2). The Bacillus spp. count was lower Bacillus spp. count. High Bacillus spp. count observed
than 9 Log10 cfu/g for FD and TALB less than 10 h and during the ALB fermentation was pointed out by many
6 cm, respectively (Fig. 3b). This result can be explained

123
J Food Sci Technol
studies (Beaumont 2002; Ouoba et al. 2004). Similarly,
high Bacillus spp. count was reported during others legu-
minous seed fermentation such as soybean (Dakwa et al.
2005) and roselle bean (Agbobatinkpo et al. 2012). Legu-
minous seed fermentation is generally an alkaline fer-
mentation. For most alkaline fermented foods, raw
materials undergo a long cooking time prior to fermenta-
tion and this contributes to elimination of non-spore-
forming bacteria (Parkouda et al. 2009) favoring the pre-
dominance of Bacillus spp. during the ALB fermentation.
pH
Experimental data of pH of fermented ALB were used to
generate the following regression equation:
pH ¼ 5:6 þ 7  102 X1 þ 3  104 X21 þ 7  102 X2
þ 2  104 X22 þ 5  102 X3 þ 4  101 X23
6  103 X1 X2 þ 8  103 X1 X2 3  102 X2 X3
ðX1 ¼ FD; X2 ¼ TALD and X3 ¼ PYÞ
The model explained 97% of variation. All the independent
variables showed a significant linear effect on pH
(Table 2), with the fermentation duration (FD) and the
thickness of the layer of ALB cotyledons in fermentation
tray (TALB) being the most important independent vari-
ables. Indeed, the pH increased significantly (p \ 0.001)
with the fermentation duration whereas the inverse effect
was observed with the TALB. Whatever the FD, the pH
remained lower than 7.5 as long as the TALB is lower than
6 cm (Fig. 3c). The pH increased with the fermentation
duration (from 6.4 to 8.4) (Table 1). This observation is
consistent with previous works (Agbobatinkpo et al. 2012;
Pelig-Ba 2009 and Azokpota et al. 2006). The increase in
pH during ALB fermentation can be attributed to the
abundant production of ammonia from the desamination of
amino acids (Pelig-Ba. 2009). It has been reported that
ammonia production commonly accompanies the fermen-
tation of high protein containing food like dawadawa,
soumbala, sonru (Omafuvbe et al. 2000; Beaumont 2002,
Azokpota et al. 2006).
Penetration force (softness indices)
Penetration force (PF) expresses the softening of ALB
which is an important criterion for sonru quality. The
relationship between PF and independent variables was
established as follows:
Penetration force ¼ 323:03  26:3X1 þ 5  101 X21 9:4X2
þ 4  101 þ 148:1X3  54:3X23
þ 101 X1 X2 4:1X1 X3 þ 1:6X2 X3:
ðX1 ¼ FD; X2 ¼ TALB and X3 ¼ PYÞ
The model could be adequately used as predictor model
due to 99% of determination coefficient. The PF decreased
significantly as the fermentation progressed. The fermen-
tation duration had a negative linear (p \ 0.001) and
quadratic (p \ 0.001) effect. The proportion of yanyanku
(PY) did not show any significant effect on PF. Irrespective
of the PY and the TALB, the PF decreased 60–70% during
the first 12 h of fermentation (Fig. 3d). Accordingly, the
ALB cotyledons softness increased as the fermentation
progressed. In addition, high and positive correlation
(r = 0.85) was observed between data from ranking test
and penetration force measured. The increase of ALB
cotyledons softness during the fermentation is in agreement
with previous findings reported by Agbobatinkpo et al.
(2012) who reported that the ALB fermented using
yanyanku were significantly softer than those fermented
without yanyanku. This suggested that yanyanku plays an
important role in the softening of ALB cotyledons during
the fermentation. The ability of Bacillus subtilis to soften
the tissue of African beans has clearly been established as
the strains of species produce enzymes that readily
hydrolyzed pectin and complex protein of the beans
(Mbajunwa et al. 1998; Njoku et al. 1990). B. subtilis is
known as the main species in condiments from fermented
ALB as soumbala, dawadawa, sonru… (Ouoba et al. 2004;
Olasupo et al. 2010) and in yanyanku used as traditional
starter for sonru fermentation (Agbobatinkpo et al. 2013).
Correlation between penetration force, pH
and microorganisms count
The penetration force was highly and negatively correlated
with Bacillus spp. count (r = -0.78, p \ 0.05) and pH
(r = -0.57, p \ 0.05). Bacillus spp. was also correlated
with the pH (r = 0.47, p \ 0.1). The correlation between
penetration force and Bacillus spp. count suggested that
Bacillus spp. were strongly involved in the softening of
ALB during the fermentation. The correlation between
Bacillus spp. and the pH was expected as Bacillus spp.
were responsible of the degradation of proteins which
produced ammonia following by pH increase during ALB
fermentation.
Optimal conditions and experimental validation
The optimal processing parameters were determined in
order to maximize Bacillus spp. and AMB count and pH
123

and to minimize the penetration force. The measured val-
ues lay within a 77% mean confidence interval of the
predicted value for Bacillus spp. and AMB count, pH and
penetration force. This high coefficient (77%) shows the
predictability of the model for the ALB fermentation in the
experimental condition. The model describes the optimal
conditions for the ALB fermentation as: FD of 24 h, 100 g
of yanyanku for 100 kg of ALB (1%) and TALB of
8.5 cm. The predicted responses from these optimal con-
ditions are 10.7 log CFU/g for AMB count, 11.4 log CFU/g
for Bacillus spp. count, 7.3 for pH and 0.09 N for pene-
tration force. The predicted values of AMB count, pH and
penetration force were similar to those reported by Agbo-
batinkpo et al. (2012); nevertheless, the predicted value of
Bacillus spp. count was higher than values reported by
these authors. This difference can be explained by the fact
that in this study, the ALB fermentation was conducted in
fermentation wooden tray instead of in basket. The use of
tray for ALB fermentation would be probably improved
fermentation conditions. Experimental trials were con-
ducted in four replicates using the above optimal condi-
tions for the model validations. The experimental response
(7.4 ± 0.2, 11.5 log CFU/g ± 0.5, 11.2 log CFU/g ± 0.6
and 0.16 N ± 0.04 for pH, Bacillus spp. count, AMB count
and penetration force respectively) were similar to pre-
dicted values (7.3, 11.4 log CFU/g, 10.7 log CFU/g and
0.09 N for pH, Bacillus spp. count, AMB count and pen-
etration force respectively).
Penalty analysis based on JAR test to identify
drivers of liking and directions for sonru
Consumer acceptability of sonru
The consumer acceptance was evaluated on sonru prepared
using optimal conditions for ALB fermentation. The panel
included seventy-six (76) consumers (75% of women and
25% of men) with 18–60 years old who use fermented ALB
condiments at less once per week. The consumers included
street food sellers (31%), producers and sellers of fermented
ALB condiment (21%) and students (18%). Globally, the
consumers gave good score for acceptability of sonru. The
mean score was 6.0 using a 7-point hedonic scale (from
dislike very much = 1 to like very much = 7). They were
classified into three groups following the scores acceptabil-
ity. The group 1 (32% of consumers) gave 7.0 as score, the
group 2 (43%) and group 3 (25%) gave 6.0 and 4.5 respec-
tively. All consumer groups gave a score over 4 (neither like
nor dislike) which indicated a good acceptance of sonru.
123
J Food Sci Technol
JAR test and penalty analysis
In order to measure the deviation from ideal levels per
attribute (Agudelo et al. 2015), the Just-About-Right (JAR)
test was performed on sonru presented on two forms:
(a) plain sonru and (b) sonru incorporated into cooked rice.
Sonru was closed ideal as regard to color (80% of the
consumers), odor (66%), flavor (64%) and taste (60%).
This could be explained considering the existence of a halo
effect. As discussed by Ares et al. (2009), when consumers
like a product, they tend to like all the liking attributes
considered and vice versa. However, only 42% of con-
sumers reported that the softening was closed ideal versus
43% that judged too soft.
Penalty analysis was performed to identify potential
directions for future improvement of organoleptic quality
of sonru (Agudelo et al. 2015). Based on the Pareto prin-
ciple, the 20% cutoff theory on the percentage of not-JAR
consumer is used as general rule for penalty analysis
(Plaehn 2012; Agudelo et al. 2015). Figure 4a shows how
many points of liking were lost for having a sonru not JAR
(too strong or much and too weak or low depending on
variables). Irrespective of sonru form (plain or with rice),
all attributes received significant and higher decreases
(0.8–1.3) in overall liking when they were judged too
weak; but the softening and color too weak was reported by
less than 20% of consumers. However, the odor too weak
was reported by more than 20% of consumers. This weak
odor can be explained by the fermentation duration which
was 24 h according to the optimal conditions. Indeed,
traditionally, ALB seeds were fermented for 48 h during
sonru production. Previous studies revealed that concen-
tration of most of the volatile compounds identified in ALB
fermented condiments increased with the fermentation time
(Azokpota et al. 2010). Concerning the characteristic ‘‘too
much’’, only the taste significantly decreased in overall
liking, as observed by less than 20% of consumers. Thus,
taste fell under the bipolar condition (i.e., a portion of the
population considered significantly the level ‘‘too weak’’
and portion of the population thought it was significantly
‘‘too strong’’). This can be explained by the fact that sonru
in cooked food is a versatile commodity because its sen-
sory characteristics can be adjusted to conform to con-
sumer demand/specifications. In addition, the proportion of
sonru used to prepare the rice for the test probably
explained values observed. Figure 4b summarizes the
results from penalty analysis which is used in sensory data
to identify potential directions for product improvement.
Sonru has been highly appreciated by consumers and the
selected sensory attributes have not greatly penalized the
overall acceptance. Indeed, the penalty varies from 0 to 0.3.
In their study on wine, Cadot et al. (2010) reported the
penalties from 0.1 to 1.2 and they considered the
J Food Sci Technol
(a)
No significant
Penalty significant and less than 20% consumers
Penalty significant and more than 20% consumers
Fig. 4 Mean drops for the ‘‘too strong’’ and ‘‘too weak’’ levels (a) and representation of the significant penalties according to the proportion of
consumers (b)
descriptors highly penalized the acceptance when the
penalty was higher than 0.5 and reported by more than 30%
of the consumers. Horne et al. (2010) reported that in the
case where the overall acceptance was evaluated using
9-hedonic scale, the descriptors with penalty higher than
0.3 should highly penalize the overall acceptance. In our
study, we used 7-hedonic scale and the descriptor that
penalized the overall acceptance was mainly aroma
because the penalty was about 0.3 for 30% of consumers.
Conclusion
The response surface methodology (RSM) design used
highlighted the optimal conditions for ALB fermentation
for sonru production, which can be set at 24 h of fermen-
tation duration, 1% of yanyanku (100 g of yanyanku for
100 kg of ALB) and 8.5 cm for the TALB. The derivative
sonru was sensory scored by consumers with the mean
score of 6.0 using a 7-point hedonic scale. Most of con-
sumers (60–80%) found that the sonru was very good
(JAR) based on intensity of attributes except for the soft-
ening of the sonru. 30% of consumers considered that the
odor of sonru was significantly too weak, thus providing
direction in sonru improvement. Further investigation on
odor of sonru and the drying conditions of sonru for better
conservation of the product in general and the organoleptic
parameters specifically would be necessary.
Acknowledgements The authors would like to thank the Interna-
tional Foundation for Sciences (IFS) (Grant No. 5849-1) for his
financial support.
(b)
References
Agbobatinkpo BP, Azokpota P, Akissoe N, Kayodé P, da Gbadji R,
Hounhouigan DJ (2011) Indigenous perception and characteri-
zation of Yanyanku and Ikpiru: two functionnal additives for the
fermentation of African locust bean. Ecol Food Nutr
50(2):101–114
Agbobatinkpo PB, Dabadé SD, Laleyè F, Akissoe N, Azokpota P,
Hounhouigan DJ (2012) Softening effect of Ikpiru and
Yanyanku, two traditional additives used for the fermentation
of African locust bean (Parkia biglobosa) seeds in Benin. Int J
Biol Chem Sci 6(3):1281–1292
Agbobatinkpo PB, Thorsen L, Nielsen D, Azokpota P, Akissoe N,
Hounhouigan DJ, Jakobsen M (2013) Biodiversity of aerobic
endospore-forming bacterial species occurring in Yanyanku and
Ikpiru, fermented seeds of Hibiscus sabdariffa used to produce
food condiments in Benin. Int J Food Microbiol 163:231–238
Agudelo A, Varela P, Fiszman S (2015) Fruit fillings development:
multiparametric approach. LWT Food Sci Technol 61:564–572
Ares G, Barreiro C, Giménez A (2009) Comparison of attribute liking
and jar scales to evaluate the adequacy of sensory attributes of
milk desserts. J Sens Stud 24:664–676
Azokpota P, Hounhouigan DJ, Nago MC (2006) Microbiological and
chemical changes during the fermentation of African locust bean
Parkia biglobosa, to produce Afitin, Iru and Sonru, three
traditional condiments produced in Benin. Int J Food Microbiol
107:304–309
Azokpota P, Hounhouigan DJ, Annan NT, Odjo T, Nago MC,
Jakobsen M (2010) Volatile compounds profile and sensory
evaluation of Beninese condiments produced by inocula of
Bacillus subtilis. J Sci Food Agric 90:438–444
Beaumont M (2002) Flavouring composition prepared by fermenta-
tion with Bacillus spp. Int J Food Microbiol 75:189–196
Cadot Y, Caillé S, Samson A, Barbeau G, Cheynier V (2010) Sensory
dimension of wine typicality related to a terroir by quantitative
description analysis, Just About Right analysis and typicality
assessment. Anal Chim Acta 660:53–62
Dakwa S, Sakyi-Dawson E, Diako C, Annan NT, Amoa-Awua WK
(2005) Effect of boiling and roasting on the fermentation of
123

soybeans into dawadawa soy-dawadawa. Int J Food Microbiol
104:69–82
El Sheikha AF (2018) Revolution in fermented food. From Artisan
household technology to era of biotechnology. In: El Sheikha
AF, Levin RE, Xu J (eds) Molecular techniques in food biology:
safety, biotechnology, authenticity and traceability. Wiley,
Chichester, pp 241–260
El Sheikha AF, Montet D (2014) African fermented food: historical
roots and real benefits. In: Ray RC, Montet D (eds) Microor-
ganisms and fermentation of traditional foods. Food biology
series. CRC Press Taylor & Francis, Boca Raton, pp 248–282
Horne J, Plaehn D, and Stucky G (2010) Understanding penalty
analysis. http://www.marketresearchworld.net/index.php?option=
com_content&task=view&id=3233&Itemid=74. Accessed July
2018
Kishida M, Seike Y, Kawasaki H (2009) Identification and charac-
terization of a psychrophilic yeast strain newly isolated from
fermentative starter (Loog-pang) of a traditional drink in
Thailand. Biocontrol Sci 14:119–122
Krishnaiah D, Sarbatly R, Nithyanandam R (2011) Optimization of
spray for drying Morinda citrifolia L. fruit extract. J Appl Sci.
https://doi.org/10.3923/jas.2011
Mbajunwa OK, Akingbala JO, Mulongoy K, Oguntimein G (1998)
Starter culture evaluation for the production of ugba from
African oil bean seed Pentaclethra macrophylla. J Sci Food
Agric 77:127–132
Myers RH, Montgomery DC, Anderson-Cook CM (2009) Response
surface methodology, process and product optimization using
designed experiments. Wiley, Hoboken
Njoku HO, Ogbulie JN, Nnubia C (1990) Microbiological study of the
traditional processing of African oil bean Pentaclethra macro-
phylia, for ugba production. Food Microbiol 7:13–26
Nout MJR, Rombouts FM, Havelaar A (1989) Effect of accelerated
natural lactic fermentation of infant food ingredients on some
pathogenic microorganisms. Int J Food Microbiol 8:355–361
Odunfa SA (1988) Review: African fermented foods: from art to
science. MIRCEN J 4:259–273
Olasupo NA, Odunfa SA, Obayori OS (2010) Ethnic African
fermented foods. In: Tamang JP, Kailasapathy K (eds) Fer-
mented foods and beverages of the world. CRC Press/Taylor &
Francis, FL, pp 323–352
123
J Food Sci Technol
Omafuvbe BO, Shonukan OO, Abiose SH (2000) Microbiological and
biochemical changes in the traditional fermentation of soybean
for ‘‘soy-daddawa’’ Nigerian food condiment. Food Microbiol
17:469–474
Ouoba LII, Rechinger KB, Diawara B, Traore AS, Jakobsen M (2003)
Degradation of proteins during the fermentation of African
locust bean (Parkia biglobosa) by strains of Bacillus subtilis and
Bacillus pumilus for production of Soumbala. J Appl Microbiol
94:396–402
Ouoba LLI, Diawara B, Amoa-Awua WK, Traore AS, Moller PL
(2004) Genotyping of starter cultures of Bacillus subtilis and
Bacillus pumilus for fermentation of African locust bean Parkia
biglobosa, to produce soumbala. Int J Food Microbiol 90:197–205
Ouoba IIL, Thorsen L, Varnam HA (2008) Enterotoxins and emetic
toxins production by Bacillus cereus and other species of
Bacillus isolated from Soumbala and Bikalga, African alkaline
fermented food condiments. Int J Food Microbiol 124:224–230
Parkouda C, Nielsen DS, Azokpota P, Ouoba LII, Amoa-Awua WK,
Thorsen L, Hounhouigan JD, Jensen JS, Tano-Debrah K,
Diawara B, Jakobsen M (2009) The microbiology of alkaline-
fermentation of indigenous seeds used as food condiments in
Africa and Asia. Crit Rev Microbiol 35(2):139–156
Pelig-Ba KB (2009) Effect of Ash, KOH and Millet on the
fermentation of Parkia biglobosa seeds to form condiment.
Pak J Nutr 8(10):1548–1554
Plaehn D (2012) CATA penality/reward. Food Qual Prefer
24:141–152
Sneath PH (1986) Endospore-forming gram-positive rods and Cocci.
In: Sneath HP, Mair SN, Sharpe ME, Holt GJ (eds) Bergey’s
manual of systematic bacteriology. vol 2. Williams & Wilkins,
Baltimore, London, pp 1104–1137
Ying B, Gaowa S, Wenli H (2014) Response surface methodology
(RSM) in evaluation of the vitamin C concentrations in
microwave treated milk. J Food Sci Techol. https://doi.org/10.
1007/s13197-014-1505-5
Zheng XW, Tabrizi MR, Nout MJR, Han BZ (2011) A traditional
Chinese liquor fermentation starter. J Inst Brew 17(1):82–90
Publisher’s Note Springer Nature remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations.