Journal of Water Process Engineering 6 (2015) 193202

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Journal of Water Process Engineering

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Statistical optimization of Basic Blue 41 dye biosorption by

Saccharomyces cerevisiae spent waste biomass and photo-catalytic
regeneration using acid TiO2 hydrosol
Nour Sh. El-Gendy a, , Radwa A. El-Salamony a , Salem S. Abu Amr b , Hussein N. Nassar a
a
b

Egyptian Petroleum Research Institute, Nasr City, Cairo 11727, Egypt

School of Civil Engineering, Engineering Campus, Universiti Sains Malaysia, Nibong Tebal, Penang 14300, Malaysia

a r t i c l e

i n f o

Article history:
Received 29 November 2014
Received in revised form 11 April 2015
Accepted 14 April 2015
Keywords:
Biosorption
Photo-catalytic degradation
Basic Blue 41
Saccharomyces cerevisiae
Acid TiO2 hydrosol

a b s t r a c t
Factorial experiments with ve factors; stirring rate, process time and temperature, initial dye concentration and biosorbent dosage at three levels were conducted based on central composite design of
experiments to investigate their effect on Basic Blue 41 dye biosorption onto bioethanol fermentation
spent waste biomass of Saccharomyces cerevisiae. A highly statistically signicant quadratic model at 95%
2
condence level (p < 0.0001,R2 0.9612andRadj
0.9386) was developed to charcterize the inuence of these
variables on biosorption effeciency. Response surface methodology was employed to optimize the process, recording maximum biosorption % of 94% (23.5 mg/g) under static condition, within 14 h at 20 C
using 0.6% biosorbent in an initial dye solution of 150 mg/L. Approximately 92% of adsorbed dye was desorbed by elution with self-clean acid TiO2 hydrosol (pH 2) and the regenerated biosorbent was employed
for four successive cycles. The photo-catalytic degradation of the desorbed dye under ultraviolet illumination (8 W) followed the pseudo rst order kinetic model (R2 0.9687) with apparent rate constant Kapp
of 0.0043 min1 . The proposed integrating biosorption with self-clean desorption and photo-catalytic
degradation process, resulted in no secondary pollution in the form of any concentrated wastes, thus has
important environmental and economic aspects.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
Discharge of industrial efuents contaminated with dyestuffs
in the waterways is a matter of concern for both toxicological
and esthetical reasons [1]. Azodyes are the mostly used colorants
(6070%), the chemical groups constructing these dyes; azo group
( N N ), aromatic rings and auxo-chromes ( OH, SO3 , etc.) make
their bioremediation from efuents a sophisticated matter [2].
The conventional methods for treating dye-containing wastewater; occulation, coagulation, precipitation, membrane ltration,
ozonation, elctrochmical techniques and biodegradation are usually ineffective for complete removal of these compounds [3]. The
most widely used and effective method is adsorption using activated carbon, zeolite, grapheme oxide and nano-porpus silica, etc.,
but its running costs are expensive [4,5]. Implementation cost of
an efuent treatment plant is of prime consideration inuencing
decision-making; this led many researchers to search for alterna-

Corresponding author. Tel.: +20 1001443208.

E-mail address: nourepri@yahoo.com (N.Sh. El-Gendy).
http://dx.doi.org/10.1016/j.jwpe.2015.04.007
2214-7144/ 2015 Elsevier Ltd. All rights reserved.

tive readily available and cheap biomass for biosorption application

[6].
To minimize the process cost and the disposal problem of
pollutant-loaded biosorbents; priceless spent waste biosorbent as
by-products of other industrial processes can be applied in biosorption technique and used biosorbents should be regenerated and
reused repeatedly. Regeneration of cationic dye-loaded biosorbents
with dilute acid solution is well known [7], but these regeneration methods leave concentrated solutions of dyes, which bring a
secondary pollution to the environment that should be eliminated.
In this study; response surface methodology RSM as an assemblage of central composite design of experiments and multiple
regression-based methods was applied to study the inuence of
stirring rate, process time and temperature, initial dye concentration, and biosorbent dosage on the biosorption of a recalcitrant
stable textile industry widely used water-soluble cationic azo-dye;
Basic Blue-41 (BB-41). In an aspect of re-use of one of the waste
by-products of industrial production process to treat industrial
wastewater efuent in a cost effective process; bioethanol fermentation spent waste biomass of Saccharomyces cerevisiae was used
as a biosorbent. A self-clean eluent-acid TiO2 hydrosol with a high

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N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

Table 1
Experimental range and levels of independent process variables for BB-41 removal.
Range and levels
Independent variables

+1

Stirring rate (A, rpm)

Process time (B, h)
Process temperature (C, C)
Initial dye concentration (D, mg/L)
Biosorbent dosage (E, % w/v)

0
4
15
100
0.2

50
12
25
150
0.4

100
20
35
200
0.6

Fig. 1. Chemical structure of Basic Blue 41 dye.

photo-catalytic activity was prepared and rstly used as an eluent to desorb BB-41 from the sorbent, and then its photo-catalytic
degradation capabilities on the desorbed dye was studied, in an
attempt that, the hydrosol could be continuously used in desorption
and photo-degradation process, which consequently would economize large volume of the eluent and would not bring secondary
pollution.
2. Materials and methods
2.1. Biosorbent
Fermentation spent waste biomass of S. cerevisiae, purchased
from Petroleum Biotechnology lab, Egyptian Petroleum Research
Institute was used in this study.
The biomass was washed three times with distilled water, dried
overnight at 60 C, then grinded in a mortar, sieved to constant size
(0.070.08 mm) and stored for further use.
2.2. Adsorbate
The dye used in this study was, C.I. Basic Blue 41 (Synonyms: Panacryl; Blue X-GRL; Maxilon Blue GRL 300%; Sevron
Blue GR; Kayacryl Blue GRL; Anilan Blue GRL; Synacril Blue
G; Abcol Blue GRL 900%; Aizen Cathilon Blue GRHL; Basacryl
Blue Z-3GL or Astrazon Blue FGGL) is a cationic azo textile dye, and was purchased from Ciba Specialty Chemicals Inc. [color index C.I. number: 11,105, C.B. number:
CB0499107, molecular formula: C20 H26 N4 O6 S2 , molecular weight:
482.57, IUPAC name: 2-[N-ethyl-4-[(6-methoxy-3-methyl-1,3benzothiazol-3-ium-2-yl)diazenyl]anilino]ethanol;methyl sulfate.
It is also named as benzothiazolium, 2-((4-(ethyl(2-hydroxyethyl)
amino)phenyl)azo)-6-methoxy-3-methyl-, methyl sulfate (salt).
The structure of Basic Blue-41 (BB-41) is illustrated in Fig. 1.
A stock solution of BB-41 with a concentration of 200 mg/L was
prepared in distilled water, which was further diluted according to
the experimental conditions.
2.3. Analysis of dye
The max 608 nm of BB-41 was determined on a doublebeam JASCO UV/Vis/NIR spectrophotometer model V-570 (JASCO
Analytical instruments, 8649 Commerce Drive, Easton, Maryland
21,601-9903, USA). Standard curve for different concentrations of
the dye solution (2.5100 mg/L) was established as the concentration range of BB-41 adhering to Beers law within 0100 mg/L and
distilled water was used as the blank.
2.4. Batch biosorption experiments
Each batch adsorption experiment was conducted by contacting
100 mL (pH 7) of different initial concentrations of BB-41 (adsorbate) with known dose of dried biomass as an adsorbent in 250 mL
Erlenmeyer asks closed with PARAFILM M to prevent evaporative loss and placed in a rotary shaking incubator set at different

speeds and temperatures according to the experimental conditions.

Adsorbent was separated from the solution at predetermined time
intervals by centrifugation at 2000 rpm for 15 min, and then collected for the regeneration experiments. The absorbance of the
supernatant solution at 608 nm was measured to determine the
residual dye concentration and to calculate the percentage of dye
removal. Negative controls (with no biosorbent) were carried out
to ensure that any sorption effect of dye onto the wall of the conical
asks is ruled out. The dye concentration of the control was used as
the initial concentration (Co mg/L) for calculations of dye removal,
using the following equation:
Dye removal =

C C 
o
t
Co

100

(1)

where, Ct mg/L is the residual dye concentration at different time

intervals (t,h). All experiments were conducted in triplicates and
the listed data are the average of the obtained results, with a standard deviation SD of 0.5.
2.5. Factorial design of experiments
Response surface methodology (RSM) was used to optimize and
investigate the inuence of different process variables on the batch
biosorption process. The central composite design CCD was applied.
The experimental runs were carried out according to a 25 full factorial design for ve independent variables; stirring rate (A,rpm),
process time (B,h), process temperature (C, C), initial dye concentration (D, mg/L), and biosorbent dosage (E, % w/v), with low (1)
and high (+1) levels. The total number of experiments is given by
the following formula:[50 = 2k + 2 (k) + 8], where k is the number
of independent variables (k = 5), this includes; 32 factorial point
from 42 full factorial CCD augmented with 8 replicates at the center point to assess the pure error. The response selected, was the
percentage dye removal. The levels were selected, based on preliminary study results. The design factors with low (1), high (+1) and
center (0) levels for the experimental design are listed in Table 1.
2.6. Statistical analysis
Once the experiments were preformed, the next step was to perform a response surface experiment to produce a prediction model
to determine curvature, detect interactions among the design factors (independent variables), and optimize the process, that is,
determine the optimum values of independent variables leading to
high biosorption percentage. The model used in this study to estimate the response surface is the quadratic polynomial represented
by the following equation:
Y = 0 + ni=l i xi + n1
nj=i+1 xi xj + ni=l ii xi2
i=1
ij

(2)

where, Y is the dye percentage removal, n is the number of factors,

0 is the intercept term, i , ij and ii are the linear, interactive and
quadratic coefcients, respectively, while, xi  s are the levels of the
independent variables (factors) under study.
The statistical software Design Expert 6.0.7 (Stat-Ease Inc., Minneapolis, USA) was used for design of experiments, regression and

N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

195

Table 2
The central composite design matrix for ve coded independent variables along with experimental and predicted BB-41 dye removal percentage.
Run
number

A
Stirring rate (rpm)

B
Time
(h)

C Temperature
( C)

D
Initial dye
concentration (mg/L)

E Biosorbent
dosage % (w/v)

Dye removal %
Experimental

Predicted

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50

1
1
1
0
+1
0
0
1
1
1
1
+1
+1
1
0
+1
+1
+1
+1
0
+1
1
0
+1
0
+1
+1
0
+1
+1
1
1
0
0
1
0
0
+1
1
0
1
0
1
1
+1
0
1
+1
+1
0

1
1
0
0
+1
+1
0
1
1
1
+1
+1
+1
1
0
+1
1
0
1
0
+1
+1
0
1
0
1
1
0
+1
1
+1
1
0
0
+1
0
1
+1
1
0
+1
0
+1
+1
1
0
+1
+1
1
0

+1
+1
0
0
0
0
0
1
+1
1
1
1
+1
+1
0
1
+1
0
1
0
+1
1
+1
1
0
+1
+1
0
+1
+1
+1
1
0
0
+1
0
0
1
1
0
+1
0
1
1
1
0
+1
+1
1
1

1
+1
0
0
+1
0
0
1
+1
+1
1
1
+1
1
0
+1
1
0
+1
0
-1
-1
0
-1
-1
+1
-1
0
-1
+1
+1
-1
0
+1
+1
0
0
1
+1
0
-1
0
+1
+1
-1
0
-1
+1
+1
0

+1
1
0
0
+1
0
0
+1
+1
1
1
+1
1
1
+1
1
+1
0
+1
1
1
+1
0
+1
0
+1
1
0
+1
1
+1
1
0
0
1
0
0
1
+1
0
+1
0
+1
1
1
0
1
+1
1
0

58.49
46.25
92.00
88.33
93.11
84.50
88.00
74.37
72.67
71.49
83.84
72.87
82.48
59.22
86.10
87.50
61.41
95.69
91.56
84.62
88.86
77.78
78.44
65.35
79.45
77.85
61.29
88.23
90.14
68.0
86.33
73.57
88.13
87.48
69.56
88.48
71.39
77.66
85.81
88.33
91.71
88.33
83.19
76.14
74.70
88.36
88.12
95.32
84.16
83.85

62.16
49.84
90.25
87.13
90.19
85.50
87.13
71.88
68.20
75.06
80.92
73.13
85.65
57.40
89.02
84.86
62.01
96.24
92.61
83.98
91.09
78.68
80.05
66.06
82.23
79.68
63.29
87.13
89.43
67.35
85.85
73.74
87.13
86.98
67.87
87.13
72.95
81.41
86.81
87.13
89.30
87.13
84.11
72.75
73.96
87.13
84.93
97.60
86.90
84.52

graphical analyses of the data obtained, and statistical analysis of

the model to evaluate the analysis of variance (ANOVA).
2.7. Preparation of acid TiO2 hydrosol
The TiO2 hydrosol was prepared according to Wang et al.
[8]; 1 mL titanium iso-propoxide was dissolved in 10 mL absolute
ethanol, and then the solution was added drop-wise into 30 mL
doubly distilled water under a vigorous stir. After that, the suspension was kept stirring under 70 C for about 45 min to ensure
complete hydrolysis. Then, 100 mL 0.04 mol/L HNO3 was added,
and the mixture was continuously stirred at 70 C for 4 h in an
air proof condition. Transparent hydrosol was obtained, and it was
diluted with water to 100 mL. The pH value of the hydrosol was
about 2. The obtained hydrosol was examined by a double-beam
JASCO UV/vis/NIR spectrophotometer model V-570 and transmission electron microscopy (TEM, Jeol Jem 2100F, 80 to 200 kV,

Japan). The phase identication was done using dispersive Raman

spectrometer (BRUKER-SENTERRA, Germany) equipped with an
integral microscope (Olympos). The particle size of TiO2 was determined using dynamic light scattering DLS technique (Zetasizer
version 6.32, Malvern Instruments Ltd., UK).
2.8. Biosorbent regeneration
The dye-loaded biomass was added into 20 mL of the acid TiO2
hydrosol. After stirring at room temperature for 20 min, most of
the adsorbed dyes were desorbed and transferred into the acid
TiO2 hydrosol solution system. The biomass was then separated
from the hydrosol by centrifugation, and washed in turn with NaOH
solution (0.01 M) and distilled water. The regenerated neutralized
biomass was then used in the next adsorption experiment. The
TiO2 hydrosol containing the desorbed dye was used in the next
photo-degradation experiment.

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N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

2.9. Photo-degradation of BB-41

The photo-catalytic reaction was carried out in a cylindrical
Pyrex reactor. The TiO2 hydrosol containing BB-41 was irradiated
directly under UV 254 nm (8 watt) VILBER-LOURMAT T-8C lamp at
room temperature 25 1 C. Samples were collected at prescribed
time intervals to determine photo-degradation rate of the dye by
measuring the absorbance at max 608 nm.
3. Results and discussion
3.1. Elucidation and validation of regression model
The CCD has the advantage that it permits the use of relatively
few combinations of variables for the determination of complex
response function [9]. The statistically designed experiments were
used to obtain a quadratic model consisting of 32 trials plus 8 center points involving ve independent important variables; stirring
rate rpm (A), process time h (B), process temperature C (C), initial dye concentration mg/L (D), and biosorbent dosage wt%, w:v
(E), affecting the percentage dye removal. The objective of the
data analysis is to t a regression model equation, specify regression coefcients and recognize the signicant model terms, and
nally determine the factors optimum levels which would lead to a
maximum response, i.e. maximum percentage of dye removal. The
obtained experimental results were evaluated and a second order
polynomial regression model equation relating the percentage dye
removal and process parameters was developed and represented
as follows:
Y = 87.82 + 2.19A + 7.18B 2.46C + 2.44D + 2.75E + 6.48A2
4.78B2 6.22C 2 3.90D2 2.00E 2 + 0.050AB + 1.22AC
+ 3.02AD 1.31AE + 5.28BC 2.49BD 0.29BE 2.51CD
+1.64CE + 3.69DE

(3)

Where, positive sign in front of the terms indicates synergetic effect,

whereas negative sign indicates antagonistic effect.
Table 2 illustrates the statistical combinations of variables with
the experimental and predicted response values.
It was observed that within the studied variables range, the percentage dye removal increased with increase of adsorbent dosage.
This may be due to the increase in the available active surface sites
of the adsorbent, i.e. increase in active surface area. At low biosorbent concentration; <0.4 wt%, the biosorption percent decreased
with increase of initial dye concentration, due to the saturation of
sorption sites on biosorbent. But at higher biosorbent concentration; >0.4 wt%, the biosorption percent increased with increase in
the initial dye concentration, this might be attributed to the large
number of available vacant sites on the biosorbent surface for BB41 molecules, which consequently would increase the driving force
of the concentration gradient between adsorbate in solution and
adsorbate in the adsorbent [10,11]. The increase in stirring rate
increased to some extent the dye uptake especially at higher initial dye concentration, indicating more effective contact between
biomass and dye solution at higher stirring rate (100 rpm). Ong et al.
[12] reported that, a boundary layer is surrounding the biomass
and a decrease in its effect is observed with increasing the stirring
rate. The adsorption of BB-41 on spent waste biomass of S. cerevisiae decreased as the temperature increased from 2535 C, this
indicates that with rise in temperature, afnity of the binding sites
on the biomass for the dye decreases and the process seems to be
exothermic in nature.
The validity of the tted model was evaluated and its statistical signicance was controlled by F-test. The analysis of variance
(ANOVA) for the response surface full quadratic model is given in

Table 3. It can be indicated that the model is highly statistically signicant at 95% condence level, with F-value of 36.3 and very low
probability p-value of <0.0001, i.e., there is less than 0.01% chance
that this error is caused by noise. The values of the determination
2 which measure the model tting reliabilcoefcients, R2 andRadj
ity for model (Eq. (3), were calculated to be 0.9612 and 0.9386,
respectively. This suggests that, approximately 96.12% of the variance is attributed to the variables and indicated a high signicance
of the model. Thus, only 3.88% of the total variations cannot be
explained by the model which ensures the good adjustment of the
above model to experimental data. Conrmation of the adequacy
of the regression model was reected also by the good agreement
between experimental and predicted values of response variables
as shown in Table 2. Where, the actual experimental biosorption
percentage ranged from 46.25 to 95.69% and its corresponding predicted values are 49.84 and 96.24%, respectively. Adeq Precision
measures the signal to noise ratio. A ratio greater than 4 is desirable
[13]. The ratio of 28.42 indicated an adequate signal. This model is
reliable and can be used to navigate the design space. The standard
deviation SD and coefcient of variance were low recording 2.8 and
3.46, respectively.
The relationship between the predicted and experimental values of biosorption efciency (% dye removal) is shown in Fig. 2a. It
can be seen that there is a high correlation (R2 = 0.9701) between
the predicted and experimental values indicating that the predicted and experimental values were in reasonable agreement. It
means that the data t well with the model and give a convincingly good estimate of response for the system in the experimental
range studied.
Fig. 2b shows the normal probability plots of the standardized residuals for biosorption efciency. The standardized residuals
measure the number of standard deviations separating the actual
and predicted values. A normal probability plot indicates if the
residuals follow a normal distribution, in which case the points will
follow a straight line. Since some scattering is expected even with
the normal data, as shown in Fig. 2b, it can be assumed that the
data is normally distributed. Thus, indicates a good validity for the
approximation of the quadratic regression model.
Fig. 2c shows standardized residual versus predicted values for
% dye removal. In this research, points of observed runs were scattered randomly within the constant range of residuals across the
graph. Thus, it revealed no obvious pattern and unusual structure.
i.e., the model is adequate and there is no reason to suspect any
violation of the independence or constant variance assumption in
all runs. The standardized residuals versus run plot represented in
Fig. 2d, shows randomly scattered points ranged between 3; the
errors were normally distributed and insignicant.
Analysis of variance (ANOVA) of the regression model was
carried out to nd the statistical signicance of the main and interacting effects of different studied parameters on the biosorption
process at 95% condence level. The signicance of each coefcient was determined by F-values and p-values (Table 3). The larger
the magnitude of the F-value and the smaller the p-values, the
more signicant is the corresponding coefcient. This implies that
the main effect of stirring rate, process time, initial dye concentration and biosorbent dosage have a highly positive statistically
signicant effect on dye removal (p < 0.0001), i.e., high dye removal
efciency occurs at high levels of those factors. But process temperature has a highly negative statistically signicant effect on
dye removal (p < 0.0001), i.e., a reduction in dye removal efciency
occurs at high temperatures. It is difcult to explain clearly the
adsorption process with respect to a single factor, hence, overall
interactive effects of various studied parameters were considered
in the model to determine the dye removal response and optimization of multiple interacting factors was done to improve the

N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

197

Fig. 2. Validation of model.

biosorption process. The positive interactive effect of stirring rate

and process time and the negative interactive effect of process time
and biosorbent dosage have a statistically non-signicant effect on
dye removal at 95% condence limits (p > 0.1). The positive interactive effect of stirring rate and process temperature and the negative
interactive effect of stirring rate and biomass dosage have a statistically possible-signicant effect on dye removal at 95% condence
limits (p < 0.1). The positive interactive effect of the process temperature and biosorbent dosage has a statistically signicant effect
on dye removal at 95% condence limits (p < 0.01). But the positive interactive effect of stirring rate and initial dye concentration,
process time and temperature and that of initial dye concentration and biomass dosage have highly statistically signicant effect
on dye removal (p < 0.0001). While the interactive effect of process
time and initial dye concentration and that of process temperature
and initial dye concentration have highly statistically signicant
negative effect on dye removal (p < 0.0001).

The perturbation plot Fig. 3 shows the comparative effects of

all independent variables on the percentage dye removal (biosorption efciency). The curvatures show that within the studied range
of parameters, the sensitivity of the dye removal toward the positive synergetic effect of independent variables can be ranked in the
following decreasing order process time > biomass dosage initial
dye concentration stirring rate. But high levels of process temperature have antagonistic effect on dye removal.

3.2. Response surface methodology to study the interactve effects

of process variables
Three-dimensional response surface graphical diagrams of the
regression Eq. (3) were plotted to understand the interactive relationship between the independent variables and % dye removal (i.e.
biosorption %).

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N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

Table 3
Analysis of variance of the tted quadratic regression model Eq. (3).
Source

SSa

dfa

MSa

F-value

p-value

Remarks

Model
A
B
C
D
E
A2
B2
C2
D2
E2
AB
AC
AD
AE
BC
BD
BE
CD
CE
DE
Residual
Pure error
Corrected total

6.00E + 003
164
1.75E + 003
206
202
257
104
56.5
95.7
37.6
9.94
0.0810
47.7
291
55.1
893
198
2.72
202
85.7
437
239
0.16
6240.74

20
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
29
7
49

300
164
1.75E + 003
206
202
257
104
56.5
95.7
37.6
9.94
0.0810
47.7
291
55.1
893
198
2.72
202
85.7
437
8.26
0.022

36.3
19.8
212
24.9
24.5
31.1
12.6
6.84
11.6
4.55
1.20
0.00981
5.78
35.3
6.67
108
23.9
0.329
24.5
10.4
52.9

<0.0001
0.000116
<0.0001
<0.0001
<0.0001
<0.0001
0.00135
0.0140
0.00196
0.0414
0.282
0.922
0.0229
<0.0001
0.0151
<0.0001
<0.0001
0.570
<0.0001
0.00314
<0.0001

Highly signicant
Highly signicant
Highly signicant
Highly signicant
Highly signicant
Highly signicant
Signicant
Possibly signicant
Signicant
Possibly signicant
Non signicant
Non signicant
Possibly signicant
Highly signicant
Possibly signicant
Highly signicant
Highly signicant
Non signicant
Highly signicant
Signicant
Highly signicant

SS: sum of squares, df: degree of freedom, MS: mean square.

Fig. 3. Perturbation plot for percentage dye removal.

Fig. 4a represents the interactive effect of stirring rate rpm and

initial dye concentration mg/L on the dye removal, at constant
process time, temperature, and biomass dosage of 12 h, 25 C and
0.4%, respectively. It is obvious from the contour lines and the RSM
plot that it is a highly statistically signicant positive interactive
effect. Where, under static conditions, the biosorption was low and
decreased at high initial dye concentration. However, the biosorption percentage increased with increase of stirring rate. Not only
this, but at high stirring rate; 100 rpm, the dye removal increased
with increase of initial dye concentration, recording maximum
biosorption percentage of 97% at 200 mg/L. Fig. 4b represents
the highly positive interactive effect of process temperature C and
time on dye removal at constant stirring rate, initial dye con-

centration and biomass dosage of 100 rpm, 150 mg/L and 0.4%,
respectively. It is obvious from Fig. 4b that biosorption percentage
increased with increase in process time and temperature reaching its maximum 97% within 2530 C and 20 h, but it decreased
with further increase in temperature. The rate of biosorption was
nearly sustained within 1420 h, indicating equilibrium. Fig. 4c
represents the RSM plot and contour lines of the highly statistically signicant negative interactive effect of process time and
initial dye concentration on biosorption process at constant stirring rate 100 rpm, process temperature 25 C and biomass dosage
0.4%, respectively. At short process time, the biosorption was low;
however, it increased with time reaching equilibrium within 12 h
and remained nearly sustained between 1420 h. The maximum
biosorption percent 97% was achieved at initial dye concentration
of 150 mg/L within 12 h and remained nearly sustained thereafter,
but further increment of initial dye concentration decreased the
biosorption percentage, even at long process time. The RSM plot
and contour lines, Fig. 4d, conrm the highly statistically negative
interactive effect of process temperature and initial dye concentration on biosorption % at constant stirring rate 100 rpm, process
time 12 h and biomass dosage 0.4%. At low temperature 15 C and
initial dye concentration 100 mg/L the biosorption was relatively
low recording 78%. But it increased with increase in temperature
reaching its maximum between 25 and 30 C and decreased with
further temperature increment. Generally, at high initial dye concentration >150 mg/L the biosorption decreased. Fig. 4e illustrates
the positive interactive effect of process temperature with biomass
dosage at constant stirring rate 100 rpm, process time 12 h and
initial dye concentration 150 mg/L. It is obvious that dye removal
was low at low biosorbent concentration and temperature, but,
increased with the increase of temperature and biomass dosage,
recording 97% dye removal at 25 C and 0.4%, respectively. However, the increase in temperature; >30 C lower the biosorption
efciency. Fig. 4f represents the interactive effect of initial dye concentration and biosorbent dosage at constant stirring rate, process
time and temperature 100 rpm, 12 h and 30 C, respectively. The
RSM plot represents a highly statistically signicant positive interactive effect, which is also revealed by the contour lines. It is obvious
that at low biomass the biosorption was low, but with the increase

N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

Fig. 4. Response surface and contour plots of biosorption process.

199

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N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

Table 4
Optimum conditions solutions for BB-41 biosorption by spent waste biomass of Saccharomyces cerevisiae.
Factors

Desirability

Number of trials

A(rpm)

B(h)

C( C)

D(mg/L)

E(wt%)

1
2
2

100
0
0

18
14
18

30
20
25

200
150
150

0.6
0.6
0.5

1
1
1

Biosorption %
Predicted

Experimental

100%
96%
96%

98%
94%
94%

Perecent error

Standard deviation

2%
2.08%
2.08%

0.023094

Fig. 5. UVvis spectrophotometer scan (a), Raman spectrum (b), size distribution number (c), TEM image (d) of the TiO2 hydrosol and SEM image of the biosorbent (e).

N.Sh. El-Gendy et al. / Journal of Water Process Engineering 6 (2015) 193202

201

Table 5
Efciency of spent waste biomass of Saccharomyces cerevisiae for biosorption of different dyes.
Dye

Operation conditions

Biosorption percentage

References

Basic Blue 41

150 mg/L initial dye concentration, 0.6 wt%

biomass dosage, 14 h process period, 20 C
process temperature, initial pH 7, under static
condition and contact solution of 100 mL.
100 mg/L initial dye concentration, 0.6 wt%
biomass dosage, 13 h process period, 15 C
process temperature, initial pH 7, 90 rpm and
contact solution of 500 mL.
120 mg/L initial dye concentration, 0.6 wt%
biomass dosage, 5 h process period, 25 C
process temperature, initial pH 7, 150 rpm and
contact solution of 100 mL.

Biosorption percentage 94%

Biosorption capacity 23.5 mg/g

This work

Biosorption percentage 62%

Biosorption capacity 51.7 mg/g

[11]

Biosorption percentage 69%

Biosorption capacity 13.8 mg/g

[14]

Reactive Red 88

Acid Red 14

of the biosorbent dosage; the percentage dye removal increased

reaching its maximum 97% at 200 mg/L initial dye concentration
and 0.6% biosorbent dosage.

3.3. Optimization of biosorption process

The optimization process was carried out to determine the optimum value of maximum percentage of dye removal (biosorption %),
using the Design Expert 6.0.7 software. According to the software
optimization step, the desired goal for each operational condition (A stirring rate, B incubation period, C process temperature,
D initial dye concentration and E biomass dosage) was chosen
within the studied range. The response (biosorption percentage)
was dened as maximum to achieve the highest performance. The
program combines the individual desirability into a single number,
and then searches to optimize this function based on the response
goal. Accordingly, the optimum working conditions with respective
predicted biosorption percentage were established, and the experimental results are illustrated in Table 4. The desirability function
value was found to be 1.000 for these optimum conditions. The
laboratory experimental results agree well with the predicted values. The standard deviation and percent error were calculated for
validation of experiments. Recording average of 0.023 and 2.05%,
respectively, indicating that process optimization by CCD was capable and reliable to optimize biosorption of Basic Blue-41 using spent
waste biomass of S. cerevisiae.
The fermentation spent waste biomass of S. cerevisiae was
reported as an effective biosorbent in different batch biosorption
processes (Table 5).

3.4. Characterization of prepared TiO2

The prepared hydrosol showed a sharp peak at 300 nm , indicating the purity of the prepared titania hydrosol (Fig. 5a). Raman
spectra of the hydrosol TiO2 sample (Fig. 5b) revealed major
peaks at 70, 465, 563, 778.7 and 800 cm1 [15]. These display the
Raman spectra of anatase and rutile TiO2 . The main bands of water
were also detected at 1095, 2380, 2414, 3242, 3384, 3427 and
3464cm1 [16]. The average size of the prepared TiO2 hydrosol
recorded 11 nm, as depicted from the statistical analyses of the
TiO2 hydrosol using DLS technique (Fig. 5c). This was also conrmed
by the TEM image of the sol particles (Fig. 5d), where a good dispersibility of the crystalline particles was obtained and the size of
the sol particles was in the range of 615 nm. The scanning electron
microscope SEM image of the yeast using SEM, JEOL-model JSM53000, Japan (Fig. 5e), showed that its diameter size was 5 m,
i.e. it is approximately several hundred times greater than that of
the sol particles. Thus it was very easy to separate and collect the
biosorbent and the photo-catalyst after the regeneration step.

3.5. Regeneration and reuse of biosorbent

In the viewpoint of energy consumption, process time and cost,
the biosorption batch process was performed under static condition, using the following selected optimum conditions; 150 mg/L
initial dye concentration, 0.6 wt% biomass dosage, 14 h process
period, 20 C process temperature. The maximum biosorption
percentage of 94% with biosorption capacity of 23.5 mg/g was
obtained. The prepared acid TiO2 hydrosol (pH 2) showed good desorption capacity. The elution efciency was about 92%, and most
of the loaded dye was desorbed from the biosorbent and transferred from the biomass surface into the acid hydrosol solution.
The functional groups on the surface of the spent waste biomass
of S. cerevisiae are carboxyl, hydroxyl and amide [17]. The surface of yeast would become positively charged at low solution pH,
thus increasing the electrostatic repulsive force between the BB-41
dye and the biomass. The biosorbent was easily separated by centrifugation, washed by NaOH solution (0.01 M) then distilled water
several times, until neutralization, and then reused in the next cycle
of adsorption experiment.
When applying the regenerated biomass for removal of the
remaining dye from the rst batch, approximately complete dye
removal occurred of 99% with total biosorption capacity of
24.75 mg/g.
Upon applying, the regenerated biomass for new three successive adsorption-desorption cycles of new batches with initial dye
concentration of 150 mg/L, biosorption % recorded, 94, 90%, and
55% with biosorption capacity of 23.5, 22.5, and 13.75 mg/g, respectively.

3.6. Photo-degradation of desorbed BB-41 with acid TiO2

hydrosol
A batch photo-catalytic-decolorization of the desorbed dye
(131 mg/L) was performed, under UV-illumination. The dye
was gradually degraded, recording 56.76% decolorization, after
120 min. The pseudo-rst-order kinetic equation (Eq. (4) was
applied to determine the photo-degradation rate of the collected
desorbed dye solution in a UV/TiO2 process.

ln

C 
o

Ct

= Kapp t

(4)

where Co and Ct denote the BB-41 concentrations at t = 0 and t = t,

respectively, and Kapp is the apparent pseudo-rst-order rate constant.
The kapp value of 0.0043 min1 was obtained from the slope
in the linear regression analysis of the photo-decolorization curve
with time (R2 0.9687).

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4. Conclusion
This study proved that factorial design analysis based on central
composite deign CCD of experiments and response surface methodology RSM are reliable and powerful tools for modeling, optimizing
and studying the interactive effects of ve important parameters
(stirring rate, process time and temperature, initial dye concentration and biosorbent dosage) in a batch biosorption process for
Basic Blue 41 dye using spent waste biomass of S. cerevisiae, that
recorded, maximum adsorption capacity of 23.5 mg/g and percentage dye removal of 94%.
The experiments showed that integration of the biosorption
with photo-catalytic oxidation using self-clean eluent; acid TiO2
hydrosol would be interesting for practical application, but detailed
techno-economic analysis should be performed. Firstly, the cationic
dyes would be removed effectively from aqueous efuents by readily available, low cost biosorbent, then the dye was desorbed and
photo-degraded by the same eluent; acid TiO2 hydrosol under UVillumination. Thus, would economize great volume of solvents and
it would not bring secondary pollution to the environment.
However, there are still more details needed to be investigated
in further study; the changes occurred on the biomass surface
after regeneration, optimization of the photo-degradation process,
reusability of the hydrosol and the applicability of this integrated
process for biotreatment of polluted water with different types of
dyes.
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