979 © IWA Publishing 2019 Water Science & Technology | 80.

5 | 2019

Coagulation/flocculation of textile effluent using a natural

coagulant extracted from Dillenia indica
Daniele Domingos Manholer, Maísa Tatiane Ferreira de Souza,
Elizângela Ambrosio, Thábata Karoliny Fornicoly de Souza Freitas,
Henrique César Lopes Geraldino and Juliana Carla Garcia

ABSTRACT

The aim of this study was to assess the efficiency of mucilage extracted from the fruit of Dillenia Daniele Domingos Manholer
Maísa Tatiane Ferreira de Souza
indica for enhancing coagulation in the treatment of textile effluent. The mucilage extraction was Elizângela Ambrosio
Thábata Karoliny Fornicoly de Souza Freitas
carried out in water at room temperature. The pH, concentration of coagulant FeCl3.6H2O, and Henrique César Lopes Geraldino
Juliana Carla Garcia (corresponding author)
concentration of mucilage solution were optimized with star-type central composite design (CCD). Department of Graduate Program in Chemistry,
We were able to analyze the synergistic effects between the FeCl3.6H2O and mucilage State University of Maringá,
Maringá-Paraná,
concentrations: the process of coagulation/flocculation (CF) for chemical oxygen demand (COD) Brazil
E-mail: jcgmoraes@uem.br
removal was more efficient at a low chemical coagulant concentration (8.00 mg L
1) and a higher
natural coagulant (NC) concentration (15.00 mg L
1). This demonstrated the potential of this
mucilage to treat textile effluents, with 67.66%% COD removal, 96.86% turbidity removal and 91.12%
apparent color reduction. The characterization of the mucilage of Dillenia indica was done using
Fourier transform infrared spectroscopy (FTIR) and solid-state cross-polarization magic angle
13
spinning carbon-13 nuclear magnetic resonance (CP/MAS C NMR), and the signals obtained
indicated the presence of polysaccharides, which are responsible for enhancing the CF process.
Key words | coagulation/flocculation, Dillenia indica, natural coagulant, response surface
methodology, textile effluent

INTRODUCTION

Industrial activities are the main polluters of water resources
because of the use of a high volume of water containing
species which are toxic or difficult to degrade, such as
colors (Garcia et al. ; Al-Ani & Li ; Wei et al.
). The textile sector is an industry in which the high con-
sumption of water and the characteristics of the effluent
generated cause great environmental concern. The intense
apparent color is the most notable visual characteristic of
the effluent generated, which is due to the colors which do
not fix into the fiber (Verma et al. ). This leads to effluent
with strong color, biochemical oxygen demand (BOD),
chemical oxygen demand (COD) and turbidity, in addition
to toxic chemical products. These not only affect the appear-
ance of the receiving surface waters, but reduce sunlight
penetration and gas solubilization in lakes, leading to dis-
solved oxygen depletion and alterations in biological cycles
that mainly influence the photosynthetic activities of the
aquatic biota (Pinheiro et al. ; Paschoal & Tremiliosi-
Filho ; Zahrim et al. ; Albuquerque et al. ).
This effluent needs to be treated, and in a worldwide
context the most commonly used technique to treat liquid
effluents is coagulation/flocculation (CF), which is not
only efficient but easy to operate (Prasad ; Aber et al.
; Khayet et al. ; Kim ).
At the coagulation stage a coagulant is added for the
neutralization of charges and formation of microflakes.
This is followed by the flocculation stage, in which the
microflakes aggregate and form larger macroflakes with
faster sedimentation (Lee et al. ; Zhao et al. ; Choy
et al. , ; Wang et al. ).
Many coagulants are used in CF, including inorganic
salts of aluminum and iron (de Souza et al. ); however,
the use of these salts generates a large volume of sludge; in
this context, there have been possible links between

doi: 10.2166/wst.2019.342

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 980 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

aluminum salts and Alzheimer’s disease; they are not known

to be the cause, but to accelerate the degenerative process
(Bhatia et al. ; Liang et al. ; Zhu et al. ). To
solve this issue, different studies have focused on natural
coagulants (NCs), such as tannin (Lopes et al. ), pine
cone extract (Hussai et al. ), Salvia hispanica (Tawak-
koly et al. ), Moringa oleifera (Camancho et al. ),
Abelmoschus esculentus (Freitas et al. ) and Cereus peru-
vianus cactus (de Souza et al. ) (in which the NC may
act as auxiliaries to the CF or be used alone). The use of
NCs has numerous advantages: the volume of sludge formed
is reduced by up to five times (Choy et al. ); the use of
organic salts is lower (Wang et al. ); they are environmen- Figure 1 | Dillenia indica fruit cut in half, the central region of the fruit has a gelatinous
tally friendly and biodegradable; and they improve CF by pulp (NC) that surrounds the seeds.

increasing the density of the flakes formed, making sedimen-

tation faster and more efficient (Yin ; Choy et al. ).
CF has some factors which may influence the process,
such as pH and dosage of coagulants, which require the
optimization of these factors to significantly increase the
efficiency of the process (Kim ). In order to maximise
the efficiency of the process, we applied the statistical tech-
nique of response surface methodology (RSM) to assess
each factor individually as well as in combination. RSM pro-
vides a better assessment of the data obtained with a smaller
number of experiments, which makes it an efficient, econ-
omic process (Neto ; Wei et al. ).
The NC chosen for this study was obtained from the
fruit of Dillenia indica, popularly known as ‘elephant
apple’ or ‘April flower’ (Muhit et al. ), as it is not
used as a significant food source or grown commercially,
in addition to its easy cultivation. The fruit is roughly
12.5 cm in diameter and contains many seeds covered in
a gelatinous pulp, from which the NC was extracted. The
compounds present in the Dillenia indica pulp that may
act in the CF process are different polysaccharides, arabi-
nogalactans, and tannins (Ghandi & Metha ). Other
studies had demonstrated their efficiency in CF (Choy
et al. ; Oladoja ).
MATERIALS AND METHODS
The textile effluent (TE) was obtained from an industrial
laundry for jeans located in the city of Astorga, Paraná in
Brazil, and was collected from the equalization tank. The
NC was extracted from mucilage, which is contained in
the gelatinous pulp present in the center of the Dillenia
indica fruit (Figure 1) About 1.0 g of pulp was soaked in
100.0 mL of water and stirred for 1 hour, after which the
separation of the mucilage from the fibers was carried out
by filtering the solution through a 500 micron stainless
steel filter (Freitas et al. ).
Mucilage characterization
The mucilage obtained from the Dillenia indica fruit was
characterized with Fourier transform infrared spectroscopy
(FTIR) using an infrared spectrometer (Bruker Vertex,
–70 V) at a resolution range of 4,000–400 cm
1 with 64
scans at 4 cm
1. The samples were pulverized before being
pressed into potassium bromide pellets (KBr, Merck) and
using CP/MAS 13C NMR), model Mercury plus 300, with
solids probe CP/MAS 7 mm – frequency of core resonance
1
H of 300.06 MHz and 13C of 75.458 MHz.
Process of coagulation/flocculation (CF) – jar test
The CF process was carried out in triplicate using a jar test
(model SP LABOR Microprocessed JT203.M/3/JT203.M/6).
A stock solution of 0.37 mol L
1 of the coagulant FeCl3.6H2O
(Synth) and stock solution of the NC extracted from the
mucilage were added to 250.0 mL of effluent from the
jeans laundry and stirred at 120 rpm (30 seconds) followed
by 20 rpm (15 min) (de Souza et al. ). In this test we
assessed the optimization of pH and dosages of the coagu-
lant FeCl3.6H2O and NC. After 60 min of stabilization, the
supernatant was collected for analysis.
Analytical methods
The following methods were used: for turbidity the nephelo-
metric method described in section 2130 B (APHA );

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 981 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

COD was determined by the colorimetric method employing the central point (Table 2). The number of experiments
closed reflux described in section 5220 D (APHA ) and was determined using the calculation: N ¼ 2k þ 2 k þ nc, in
total organic carbon (TOC) was measured according to the which N, k and nc are the number of experiments, the
high temperature combustion methodology described in number of factors, and the number of repetitions at the cen-
section 5310 B (APHA ; Greenberg et al. ) using tral point, respectively (Bajpai et al. ; Kim ; Heidari
Shimadzu TOC-L equipment. Apparent color reduction was et al. ).
evaluated by absorbance reduction (Freitas et al. ). Coded values for the high and low levels were obtained
using Equation (1) and the axial points using Equation (2):
Response surface methodology
Xi
X0
xi ¼ (1)
For this study we decided on the central composite design ΔX
(CCD) technique, which is the most popular type of p
4
ffiffiffiffiffi
23 (2)
response surface methodology. We used star-type planning,
which is an enlarged CCD planning of a factorial 23. We
also established the relationships between the COD and where xi is the coded value for each factor, Xi is the actual
TOC responses and the variable factors of pH, FeCl3.6H2O, value of an independent variable, X0 is the value at the cen-
dosage, and NC dosage, in addition to analyzing five levels tral point, and ΔX is the difference between these levels.
(Table 1) described as low (
1), high (þ1), central (0), and The data obtained were adjusted to a quadratic model
axial (
1.68 and þ1.68) (Figure 2), with each level corre- according to Equation (3) where β0 is the constant coeffi-
sponding to a coordinate in space (Martins et al. ). In cient, βi is the linear coefficient, βii is the quadratic
total, 18 experiments were conducted, including four at coefficient, and βij is the interaction coefficient. For each
variable, the individual effect on the process was investi-
gated.
Table 1 | Parameters and levels of CCD planning
Xk Xk Xn
2
y ¼ β0 þ i¼1
β i x i þ i¼1
β ii x i þ β x x þ ε (3)
1ij ij 1 j
Levels

The statistical significance of the model as well as of the

Parameter
1.68
1 0 þ1 þ 1.68
terms of the regression were assessed using analysis of var-
pH X1 2.32 3.00 4.00 5.00 5.68
iance (ANOVA), the F-test and RSM. All experimental

1
FeCl3 (mg L ) X2 0.91 5.00 11.00 17.00 21.09 data were compiled using the software Design Expert
NC (mg L
1) X3 1.59 5.00 10.00 15.00 18.41 7.1.3, beginning with the development of the planning
matrix followed by the development of the mathematical
model for the ANOVA and the optimization parameters.

RESULTS AND DISCUSSION

Characterization

We carried out the characterization of the mucilage from

Figure 2 | Schematic diagram of central composite design (CCD) and its factors X1 (pH),
X2 (FeCl3.6H2O) and X3 (NC), with four replicates at the central point.
the Dillenia indica fruit using FTIR (Figure 3) and CP/
MAS 13C NMR) (Figure 4).
The large absorption band in the 3,500–3,200 cm
1
range, with a central peak at roughly 3,400 cm
1 resulted
from the presence of –OH clusters, and the highest intensity
at 3,420 indicates the elongation of the OH of alcohol as
well as carboxylic acid groups (de Souza et al. ). The
peak at 2,900 cm
1 is characteristic of the –CH of aromatic
rings, the elongation of –OH, and the C ¼ O of carboxylic

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 982 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

Table 2 | CCD planning matrix obtaining as response removal of COD (Y1) and TOC (Y2)

1
1
Tests pH (X1) FeCl3.6H2O (mg L ) (X2) NC (mg L ) (X3) Y1 (%) current Y1 (%) predicted Y2 (%) current Y2 (%) predicted

1 3.00 (
1) 5.00 (
1) 5.00 (
1) 57 56 35 35

2 5.00 ( þ 1) 17.00 ( þ 1) 15.00 ( þ 1) 63 62 24 23
3 2.32 (
1.68) 11.00 (0) 10.00 (0) 75 74 15 17
4 3.00 (
1) 5.00 (
1) 15.00 ( þ 1) 56 57 9 8
5 4.00 (0) 11.00 (0) 10.00 (0) 65 66 31 33
6 5.00 ( þ 1) 5.00 (
1) 15.00 ( þ 1) 56 54 29 30
7 5.68 ( þ 1.68) 11.00 (0) 10.00 (0) 69 72 9 8
8 5.00 ( þ 1) 5.00 (
1) 5.00 (
1) 61 60 34 35
9 4.00 (0) 11.00 (0) 10.00 (0) 67 66 39 33
10 4.00 (0) 11.00 (0) 10.00 (0) 67 66 31 33
11 5.00 ( þ 1) 17.00 ( þ 1) 5.00 (
1) 60 57 1 1
12 3.00 (
1) 17.00 ( þ 1) 5.00 (
1) 56 54 36 34
13 4.00 (0) 11.00 (0) 18.41 ( þ 1.68) 48 48 25 27
14 4.00 (0) 21.09 ( þ 1.68) 10.00 (0) 60 62 30 32
15 4.00 (0) 11.00 (0) 10.00 (0) 64 66 33 33
16 3.00 (
1) 17.00 ( þ 1) 15.00 ( þ 1) 69 68 37 35
17 4.00 (0) 11.00 (0) 1.59 (
1.68) 41 43 31 33
18 4.00 (0) 0.91 (
1.68) 10.00 (0) 56 56 40 39

of secondary CO and OH belonging to the polysaccharides,

and the absorption peaks at 584 and 594 cm
1 correspond
to the elongation of C-OH (Rosa et al. ).
The CP/MAS 13C NMR spectrum (Figure 4) presented
enlarged peaks at 73 ppm corresponding to C-OH;
105 ppm corresponding to C-O-C, and 173 ppm corre-
sponding to the carbonyl of carboxylic acid – all these
signs indicate the presence of polysaccharides (Silverstein
et al. ) and correspond to a range of polysaccharides
present in the mucilage (Faccio ). The identification of
these groups corresponding to polysaccharides is funda-
mental for identifying possible sites of adsorption
present in the mucilage which can be active in the CF
process.

Figure 3 | FTIR spectrum of the Dillenia indica fruit mucilage.

Experimental design – CCD

acid (Silverstein et al. ). The peaks at 1,710 and
1,385 cm
1 are characteristic of the polysaccharide com-
plexes of elongation vibration of C–O and stretching
vibration of C-O-C (de Souza et al. ). The range of
800–1,200 cm
1 represents the region for the identification
of carbohydrates (Singh & Bothara ). Peaks at 1,035
and 1,070 cm
1 are attributed to the vibrational elongation
RSM was used to obtain the maximum amount of useful
information with a minimum number of experiments
(Neto ). By using RSM it was possible to assess the
effect of pH (X1), FeCl3 mg L
1(X2), and NC mg L
1 (X3)
individually, as well as the synergisticic or antagonistic
effects between the combinations of these factors. The
quadratic regression model was obtained for two

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 983 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

Figure 4 | CP/MAS 13
C NMR) spectrum of the Dillenia indica fruit mucilage.

responses, Y1 COD (%) removal and Y2 TOC (%) removal. de Souza et al. ):
Equations (4) and (5) provide the coded values for the
quadratic models; coefficients with one factor represent Y1 ¼ þ65, 65–0, 59 X1 þ 1, 81 X2 þ 1, 59 X3 –0, 75 X1 X2
the effect of that particular factor, while coefficients with
1, 75 X1 X3 þ 2, 75 X2 X3 þ 2, 65 X21 –2, 30 X22 –7, 07 X23
two factors and those with second-order terms represent R2 ¼ 96, 00%; Adjusted R2 ¼ 91, 51% (4)
the interaction between the two factors as well as the quad-
ratic effect, respectively. The positive sign in front of the Y2 ¼ þ33, 44–2, 86 X1 –1, 89 X2 –1, 25 X3 –8, 37 X1 X2
terms indicates a synergistic effect, while the negative þ 5, 38 X1 X3 þ 6, 88 X2 X3
7, 35 X21 þ 0, 78 X22 –1, 70 X23
sign indicates an antagonistic effect (Martins et al. ; R2 ¼ 96, 68%; Adjusted R2 ¼ 92, 59% (5)

Figure 5 | (a) Chart of correlation between predicted and obtained values; (b) Chart of normal probability for residues corresponding to the COD removal model.

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 984 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

Figure 6 | (a) Chart of correlation between predicted and obtained values; (b) Chart of normal probability for residues corresponding to the TOC removal model.

For COD removal, it was observed that the inorganic
coagulant and the NC had a strong influence on the effec-
tiveness of the CF process. There was a synergistic effect
when the inorganic coagulant was combined with NC, a
good result for reducing the inorganic coagulant. There
was also a synergistic effect for the quadratic effect
when using FeCl3.6H2O only. In the analysis for TOC
removal percentage, there was a synergistic effect in the
combination of pH with NC and FeCl3.6H2O combined
with NC, as well as synergy for the quadratic effect of
FeCl3.6H2O.
Statistical significance – ANOVA
These analyses were performed using the F-test and p with
a confidence level of 95% (Table 3). The value of p is used
as a tool to verify the meaning of each factor and the inter-
action between factors. If the calculated value of F (Fcal) for
the factors is higher than the tabulated value, Ftab, the
factor in question is significant, which actually occurred
for the two models studied: Fcal > Ftab and p < 0.05 at
95% confidence level.

Table 3 | ANOVA – statistical parameters obtained for the reduced models

Analysis of variance (ANOVA) – validation of the models
Sum of Mean
We validated the model using ANOVA followed by RSM, squares DF square Fcal Ftab p value

since without the ANOVA model validation the remaining COD removal

analyses could provide erroneous interpretations.

Model 1,064.15 9 118.24 21.35 3.39 0.0001
Table 2 presents the predicted and current values
Residual 44.30 8 5.54
showing that the adjusted values were close to the actual
Lack of fit 37.55 5 7.51 3.34 9.01 0.1750
values for responses Y1 and Y2. Y1 had a correlation coef-
Pure error 6.75 3 2.25
ficient (R2) of 0.9600 and Y2 indicated 0.9668, suggesting
that the empirical model was not able to explain 4.00 Total 1,108.44 17

and 3.32%, respectively. The high values of R2 indicated TOC removal

low residues with normal distributions, as illustrated in Model 2,111.94 9 234.66 25.87 3.39 <0.0001
Figures 5 and 6. The values of adjused R2. 0.9151 and Residual 72.56 8 9.07
0.9294 for Y1 and Y2, respectively, validate an optimum
Lack of fit 29.56 5 5.91 0.41 9.01 0.8186
adjustment of the experimental data to the model due to
Pure error 43.00 3 14.33
the proximity of the values of R2 and adjusted R2 (Kim
Total 2,184.50 17
).

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 985 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent
Analysis of the response surfaces for the removal of
COD and TOC
Figures 7 and 8 show the tridimensional charts of response
surfaces as function of two factors while the third factor was
maintained at its central point. For the removal of COD,
Figure 7(a) and 7(a), in which the variation in pH and
concentration of FeCl3.6H2O and NC, respectively, was
demonstrated, the best conditions for removal were the
pH values of 3 and 5. The analysis of the factors of the con-
centrations of FeCl3.6H2O and NC, in which pH was
maintained at its central point (Figure 7(c)), showed that
higher concentrations of FeCl3.6H2O did not favor COD
removal. The most significant removal occurs at a low
concentration of the coagulant FeCl3.6H2O (8.00 mg L
1)
Figure 7 | Charts of tridimensional surfaces for the interactions (a) X1X2, (b) X2X3, and (c) X1X3 as COD removal percentages.
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Water Science & Technology | 80.5 | 2019
and higher concentration of NC (15.00 mg L
1), indicating
the potential of the NC extracted from the Dillenia indica
fruit of the treatment of textile effluents.
Figure 8(a) and 8(b) illustrates the percentage of TOC
removal as function of pH as well as concentration of
FeCl3·6H2O and NC – for both the best efficiency was
obtained at pH 4.5 and concentrations of 17.00 and
4.00 mg L
1. For the removal of TOC, a chemical coagulant
concentration above 17.00 mg L
1 and a low concentration
of NC favor the process because higher NC concentrations
increase the organic matter content in the medium.
Process optimization
The software Design Expert provided 10 optimization
experiments and we chose the experiment in which a
 986 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent Water Science & Technology | 80.5 | 2019

Figure 8 | Charts of tridimensional surfaces for the interactions (a) X1X2, (b) X2X3, and (c) X1X3 as TOC removal percentages.

lower concentration of chemical coagulant presented a sat- Apparent color reduction

isfactory COD removal: pH 5.00, FeCl3.6H2O
10.72 mg L
1 and NC 9.43 mg L
1, with a COD removal Apparent color reduction from the effluent was monitored
of 67.5977%. The optimization experiment was carried out by absorbance analysis at different wavelengths, where the
in triplicate and achieved a variation coefficient below 5% biggest absorption peak in the visible region was used to
with a result of 67.66% ±2.24. monitor the apparent color reduction. This peak was
found at 670 nm, and after the software optimization pro-
cess, the percentage of apparent color reduction was
Turbidity removal 91.12%.

Using the pH, NC and FeCl3.6H2O values provided by the

optimization process, it was possible to obtain a turbidity CONCLUSION
removal of 96.86% ± 0.09. This was a very good result and
confirmed that this optimized CF process was extremely The mucilage extraction can be performed in water without
efficient. requiring the use of chemical reagents. The mucilage

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 987 D. D. Manholer et al. | Dillenia indica as coagulant of textile effluent
characterization showed peaks at 1,710 and 1,385 cm
1 cor-
responding to polysaccharides, and the CP/MAS 13C NMR)
spectrum had peaks which also correspond to a range of
polysaccharides; in the literature, CF efficiency using natural
coagulants is attributed to them.
Star-type CCD planning enabled perfect adjustment to
the quadratic model for the removal of COD and TOC
with R2 values of 91.51 and 92.59%, respectively.
High turbidity removal and apparent color reduction
were 96.86 and 91.12% respectively.
Lower NC concentrations favored the removal of COD,
demonstrating the advantage of using the Dillenia indica
mucilage in the CF process in addition to decreasing the
use of chemical coagulants, which led to the efficiency of
the technique being improved by the use of natural
coagulants.
The NC extracted from the Dillenia indica fruit proved
potentially efficient for the CF process when combined
with the inorganic coagulant. This NC had a range of advan-
tages which contribute to an environmentally compatible
process.
ACKNOWLEDGEMENTS
The authors would like to thank the State University of
Maringá for the laboratorial support and CNPq (473558/
2013-0) for the financial support. Manholer D. D., de
Souza, M. T. F.; Ambrosio, E.; Freitas, T. K. F. S., and Ger-
aldino, H. C. L., acknowledge a fellowship from CAPES/
CNPq/Fundação Araucária – program PQU.
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