Journal of Bioresources and Bioproducts.

2016, 1(1): 30-35 Peer-Reviewed

ORIGINAL PAPER

Poultry keratin based decolorants for dyeing wastewater treatment

Bi Chena,b* Long Yan a,b Xia Liua, Jessica L. Worrallc

a) School of Chemistry and Chemical Engineering, Yulin University, Yulin,719000,China.

b) Key laboratory of Industries Wastewater Treatment, Yulin University, Yulin,719000, China
c) University of New Brunswick, Fredericton, NB, Canada, E3B 5A3
*Correspondence author: Bi Chen (chenbi_110@163.com)

ABSTRACT
One of the challenges in wastewater treatment is the low efficiency in decoloring dyeing wastewater. Chicken feather, as a waste material,
has a great potential in decoloring the dyeing wastewater. In this study, a lab synthesized dyeing wastewater prepared with acid blue-A dye
was treated with a chicken feather keratin-based composite decolorant KA (keratin agent) using batch decoloration techniques. A modified
KA (MKA) was also developed to improve the decoloration efficiency. The decoloration performance of the two decolorants was then
evaluated in terms of decoloring rate, at various decolorant dosages, pH, reaction temperature and time. Under optimal conditions, the
decoloration rates of the KA and MKA in treating the dyeing wastewater were 91.8% and 94.3%, respectively. IR and TEM results
indicated that the KA and MKA decolorants removed the dye stuff from the dyeing wastewater by physical adsorption as well as chemical
reactions.

Keywords: Keratin; Poutry feather; Decolorant; Dye; Dyeing wastewater; Decoloring; Treatment

1. INTRODUCTION properties in relation to the disulphide bonds and cross-

Water pollution caused by the color effluents discharged
from textile dye manufacturers and textile dyeing mills is
one of the major environmental concerns in the world
todays.1 Textile dyeing wastewater is difficult to treat.
Color removal2 of the dyeing wastewater is especially
difficult due to its complex characteristics, large volume,
high content of organic pollutants, dark color, and unstable
properties. A number of techniques have been developed to
remove different types of dyes from wastewater.3,4 One
technique, a physicochemical method, removes different
types of dyes through adsorption,5,6 electrocoagulation,7
oxidation8 and photocatalytic decolourization9.9 Recently,
the adsorption technique has been gaining more attention
since it can be widely used in different dyeing wastewater
treatments, and it is versatile and easily to be
implimented.10 Various potential adsorbents have been
studied, such as activated carbon,11,12 clays,13 agricultural
solid wastes14,15 and fly ash.16 However, more research is
needed to develop more cost-effective dye absorbents.
Poultry Feather is a solid waste produced from
slaughterhouses. It is estimated that the annual production
of poultry feather is more than 8.5 billion tons in the
world.17 Traditionally, the majority of the feather is being
dumped into landfills, which is expensive to operate and
has harmful impacts on the environment. In addition,
poultry feather is reach in protein which breeds flies and
viruses. For these reasons, there has been an increased
interest in the utilization of feather keratin.18 For instance,
feather was used to decolor or filtrate toxic pollutants19and
dyes from dyeing wastewater.20 The poultry feather may be
activated and used directly without extracting the keratin.21
Limited studies have been found in the literature on the
extraction of keratin and its chemical and physical
linkages of feathers.
In this study, two kinds of composite decolorants were
prepared by hydrolyzing the poultry feather in sodium
hydroxide solution to extract the kerating protein which was
modified to have improved decoloring properties. The
performance of the feather keratin based decolorants in
treating a simulated dyeing wastewater was assessed by the
absorbance and decoloring rate of the treated wastewater.
The process parameters including decolorant dosage, pH,
temperature and contact time were investigated to optimize
the decoloring process. The treated dyeing wastewater,
feather keratin decolorants and resulting sludge were
examined by infrared spectrum (IR), transmission electron
microscopy (TEM), and the underlining decoloring
mechanism was discussed.
2. MATERIALS AND METHODS
2.1 Materials
Chicken feathers were collected from a slaughterhouse in
Yulin. An acid blue-A dye sample was obtained from a dye
chemical plant in Shanghai of China, and a dye-fixing agent
(DE) from Shanghai Tiantan Auxiliaries Co., Ltd.
Analytical reagent grade hydrochloric acid (HCl), sodium
dithionite (NaHSO3) and sodium hydroxide (NaOH) were
purchased from Chemicals Ltd, Xian (China).
2.2 Experimental procedures
Two different composite decolorants were prepared for
the lab trials in this study. The first decolorant (KA) was
prepared by following steps: 1) the feathers were treated
first with 40% hydrochloric acid at 75 C for 90 min; 2) and
then with 30% sodium bisulfite solution at 75 C for 90
min; 3) and then hydrolyzed with 20% sodium hydroxide

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Journal of Bioresources and Bioproducts. 2016, 1(1): 30-35
solution at 80C for 40 min. The liquid extract from the last
step contained 0.8 g/L of poultry keratin, and it was
designated as KA.
The second decolorant (MKA) was prepared by mixing
the KA (0.8 g/L) with DE (0.6 g/L) in a rotary reactor (500
mL) at 80 C for 30min, at a rotating speed of 120 r/min.
DE was a positively charged cationic polymeric quaternary
ammonium salt with an epoxy group, and the KA had polar
groups in the main chain such as -COOH, -NH2, -OH and
peptide bond (-CONH-).22,23 Thus, the epoxy group of DE
can readily react with the amino and hydroxy groups of KA
to produce MKA, which can absorb or embed dye
molecules in micelles by electrostatic attraction, hydrogen
bonding, Van der Waals' force and hydrophobic force.24
The decoloration experiment was carried out in glass
beakers with 100 mL synthesized dyeing wastewater of 60
mg/L dye concentration, according to the National Standard
Method and relative literature.16 The pH of the mixture was
adjusted to a desired level by adding hydrochloric acid or
sodium hydroxide solutions, and it was maintained at the
level throughout the whole reaction period. A PHS-3C
model pH meter was used to monitor the pH value. The
reaction temperature was also maintained at a constant level
as desired. After the decoloration treatment, the wastewater
was filtered and measured for absorbance. The absorbance
of the original and treated dyeing wastewater was
determined at 635 nm wavelength with a UV-2401PC series
spectrophotometer (SHIMADZU Co., Ltd). All experiments
were repeated three times for each condition, and the
average value was reported.
The decoloring rate of the treatment was calculated from
the change in the absorbance of the synthesized dyeing
wastewater before and after the treatment. The decoloring
rate was calculated as
nAi
R = (1 ) 100%
mA0
Where R is the decoloring rate; A0 and Ai are the
absorbance of the dyeing wastewater before and after the
treatment at 635 nm; m and n are dilution factors for the
dyeing wastewater.
3. RESULT AND DISCUSSION
3.1 Optimization of process parameters
3.1.1 Effect of pH
The pH of an adsorption process is an important process
parameter to be controlled to obtain maximum
performance.25 Figure 1 shows the effect of pH on the
decoloring rate in a wide range from pH 1 to pH 12, with
600 mg/L KA or 600 mg/L MKA, under ambient
temperature (20 C) for 30 min. For KA, the decoloring rate
maintained at about 85% between pH of 1 to 4, and
decreased dramatically to below 10% as the pH increased
from 5 to 10. This means, KA was suitable for treating
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acidic dyeing wastewater. In contrast, the results for MKA
were completely different, with the optimum performance
obtained in the neutral to alkaline range (pH from 6 to 12),
indicating that MKA was suitable for treating neutral or
alkaline wastewater. When the pH was lower than the
isoelectric point of KA, its amine groups were protonized to
impart positive charges to KA to absorb the anionic dye
molecules by electrostatic attraction. For MKA, the reactive
epoxy functional groups and cationic quaternary groups of
DE enabled it to absorb dye molecules even under basic
conditions. However, the MKA had lower efficiency in an
acidic medium as shown in Figure 1.
100
80
Decoloring rate(%)
60 KA
MKA
40
20
0
0 2 4 6 8 10 12
pH
Fig.1 Effect of pH on the decoloring rate
3.1.2 Effect of decolorant dosage
In Figure 2, the decolorant dosage was varied from 40 to
1000 mg/L for KA at pH 2, and 400 to 1600 mg/L for MKA
at pH 8, and the temperature was fixed at 20C. The results
show that the decoloring rate increased first with increasing
decolorant dosage, and then started to decrease when the
dosage was higher than 400 mg/L for the KA system, and
1400 mg/L for the MKA system. The maximum decoloring
rate was 91% and 92.8% for the KA and MKA systems,
respectively.
3.1.3 Effect of reaction temperature
Figure 3 shows the impact of reaction temperature on the
decoloring rate from 20C to 70C, with 400 mg/L KA at
pH 2, or with 1000 mg/L at pH 8. The results show that as
the temperature increased from 20 to 30 C, the decoloring
rate increased slightly, and then decreased rapidly when the
temperature increased further from 30 to 70 C. The
maximum decoloring rates were 92% for KA and 93% for
MKA, both of which were obtained at 30C. A moderately
higher temperature (e.g. 30 C) might favour the interaction
between the dye molecules and the decolorants, without
ruining the flocs formed in the absorption process.
However, when the temperature was too high (above 30
31
Journal of Bioresources and Bioproducts. 2016, 1(1): 30-35 Peer-Reviewed

C), the flocs formation might be disturbed, resulting in a

lower decoloring rate.

3.1.4 Effect of contact time 94

Figure 4 compares the decoloring rate of KA and MKA

Decoloring rate(%)
at various reaction times, in the range from 5 to 120 92
minutes. The decolorant dosage and reaction pH were fixed
at 400 mg/L and 2 for the KA system, and 1000 mg/L and 90
pH 8 for the MKA system. The results show that the
decoloring rate increased rapidly with the increase of the KA
reaction time in the first 30 minutes, and then leveled off 88
MKA
and reached a plateau at 90 minutes, which was true for
both the KA and MKA processes. 86

0 20 40 60 80 100 120
Time(min)
95
Fig.4 Effect of contact time on the decoloring rate
90
Decoloring rate(%)

3.2 Decoloring Mechanism

85
3.2.1 IR spectra
80 KA Infrared spectra of the KA and MKA decolorants are
MKA shown in curve 5 and curve 4, respectively, in Figure 5. The
absorption peaks at 3377.7 cm-1 and 1638.3 cm-1 were
75 attributed to the O-H and C=O vibration. The vibration
absorption of the C-H on the peptide chain and the N-H
70 bending vibration peaked at 1463.1 cm-1 and 1409.7 cm-1,
0 200 400 600 800 1000 1200 1400 1600
respectively, both of which intensified and shifted slightly
Dosage(mg/L) to the lower wave numbers. The absorption peak at 1650
cm-1 was responsible for the S-H groups in the spectrum for
Fig.2 Effect of MKA and KA dosages on the decoloring rate KA disappeared in the spectrum for MKA, indicating that
the dye-fixing agent DE had reacted with KA in the
decolorant preparation process.
The IR spectra for the sludge samples from the
treatments of the dyeing wastewater with KA or MKA are
95
shown by the curves 2 and 3 in Figure 4, respectively. In
KA both cases the absorption bands were intensified and
MKA
Decoloring rate(%)

90 slightly shifted to lower wave numbers. Both spectra had

the characteristic absorption peak of the acid blue A at
2359.5 cm-1, which shows that the decolorant KA and
85 MKA had interacted with the acid blue dye.8,11
The DE fixing agent had a plurality of reactive epoxy
functional groups, which can interact with the amine and
80 hydroxyl groups under alkaline conditions. The DE fixing
agent also contained multiple quaternary ammonium
75
cationic groups. It was expected that DE would crosslink
keratin and impart cationic charges to the decolorant. The
20 30 40 50 60 70
introduction of the cationic groups to the protein molecules
Temperature(C) of the keratin decolorant increased the electrostatic
attraction between the keratin decolorant and anionic dye
Fig.3 Effect of treating temperature on the decoloring rate molecules,12 and thus facilitated the absorption and
flocculation of the dye stuff in decoloration process.

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Journal of Bioresources and Bioproducts. 2016, 1(1): 30-35 Peer-Reviewed

D SO3 Na + P O CH2 CH CH2 N(R)3Cl

OH

D SO3 P O CH2 CH CH2 N(R)3 + NaCl

OH

These chemical reactions, as well as the electrostatic

attraction, Van der Waals force and hydrogen bonds
between the decolorant and the anionic dye molecules
played important roles in the decoloration of the dyeing
wastewater with MKA.

3.2.2 TEM analyses

1-Acid Blue-A; 2-MKA sludge; 3-KA sludge; 4-.MKA; 5- KA
Fig.5 The IR spectra of the decolorants and the decoloring sludge
The chemical structural of DE:
The KA decolorant particles shown in the transmission
electron microscopic (TEM) image in Figure 6a were in
separate granule form, while the MKA decolorant particles
were in cluster form (Figure 6b). The network structure of
the MKA decolorant provides further evidence for the
strong physical/chemical interaction between KA and DE.
This also explains the improved decoloring performance of
MKA in treating the dyeing wastewater. The TEM image of
the sludge from the decoloring process with MKA (Figure 6
d) had more compact structure than that from the KA
process (Figure 6c), which explains the much faster
settlement of the flocs formed in the decoloring process
with MKA than with KA.

Its simplified form:

H2C CH CH2 N(R)3Cl

O

The reactions of DE with the hydroxyl and amine groups

of KA may be described as:

P NH2 + H2C CH CH2 N(R3)Cl P NH CH2 CH CH2 N(R3)Cl

OH
O

P OH + H2C CH CH2 N(R)3Cl P O CH2 CH CH2 N(R)3Cl

O
OH

The dye (Acid Blue A) contained sulfonate groups and

was an anionic dye. The reactions of MKA with the anionic
dye may be described as follows.

D SO3 Na + P NH CH2 CH CH2 N(R)3Cl

OH

Fig.6 TEM images of KA, MKA and the corresponding sludge

D SO3 P NH CH2 CH CH2 N(R)3 + NaCl samples from the decoloration processes of the dyeing wastewater
OH with KA or MKA

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Journal of Bioresources and Bioproducts. 2016, 1(1): 30-35
4. CONCLUSIONS
Two poultry keratin-based decolorants (KA and MKA)
were successfully prepared in the lab and evaluated in
treating a dyeing wastewater synthesized with an acid blue-
A dye. It was found that the modified keratin decolorant
MKA had a superior decoloring performance than the
original keratin decolorant KA extracted from poultry
feather. The decoloring rate of the dyeing wastewater was
up to 90% with KA, and up to 92.5% with MKA, under the
conditions in current study. It was also found that the KA
was more effective in acidic conditions. While the MKA
performed better in a neutral to alkaline medium. The IR
and TEM results support the hypothesis that there were
strong physical/chemical interaction between the KA
decolorant and the DE dye fixing agent.
ACKNOWLEDGEMENT
The authors gratefully acknowledge the financial support
from Science Foundation of Department of Shaanxi
Province (2014K10-25). We are also full of gratitude for
the support provided by the Produce-learn-research project
of Yu Lin (2015CXY14-04). And thanks Ju Chen from
Polytechinique Montreal for her generous help.
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