Bioresource Technology 101 (2010) 7323–7329

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Synthesis of lignin-base cationic flocculant and its application in removing

anionic azo-dyes from simulated wastewater
Run Fang a,b, Xiansu Cheng a,*, Xiaorong Xu a
a
College of Material Science and Engineering, Fuzhou University, University Town No. 2 Xueyuan Road, Fuzhou City, Fujian Prov. 350108, China
b
Department of Chemistry and Chemical Engineering, Minjiang University, University Town No. 1 Wenxian Road, Fuzhou City, Fujian Prov. 350108, China

a r t i c l e i n f o a b s t r a c t

Article history: In this paper the Mannich reaction product of dimethylamine, acetone and formaldehyde was grafted
Received 4 February 2010 onto hydroxymethylated lignin to form a lignin-base cationic polyelectrolyte (L-DAF). L-DAF samples
Received in revised form 25 April 2010 with different cationic strengths and molecular weights were synthesized and characterized. Their per-
Accepted 29 April 2010
formance as a cationic flocculant in removing three anionic azo-dyes (Acid Black, Reactive Red and Direct
Available online 23 May 2010
Red) from simulated dye wastewater was evaluated. The removal of anionic dyes by L-DAF was ascribed
to two simultaneous mechanisms, which included coagulation by charge neutralization and flocculation
Keywords:
by bridging. There was a different optimum dose of flocculant for each dye wastewater, at which dye
Cationic flocculant
Anionic dye
removal can reach above 95%. Furthermore, a longer settling time and a lower solution pH increased
Coagulation–flocculation the efficiency of the decolorization process. During the treatment, more than 89% of chemical oxygen
Wastewater treatment demand (COD) reduction was achieved with sludge production being less than 5.4%.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction lation–flocculation process is generally used as a pretreatment

Anionic azo-dyes, which include direct, acid and reactive dyes,
are widely used due to their bright colors, their excellent color fast-
ness and the ease of application. However, wastewater containing
anionic azo-dyes usually has a strong color, a high level of toxicity
and is generally non-biodegradable. As a result, dye wastewater
represents a considerable source of environmental pollution. Due
to their complex structure and poor biodegradability, it is extre-
mely difficult to remove anionic azo-dyes from aqueous solutions.
Hence the removal of these toxic pollutants from wastewater has
rapidly become a matter of great interests and significant efforts
have been put into the development of efficient and cost-effective
treatment methods (Panswad and Luangdilok, 2000; Robinson
et al., 2001; Forgacs et al., 2004). A number of physical and chem-
ical processes are employed in decolorizing dye wastewater,
including coagulation–flocculation (Wang et al., 2008; Szygula
et al., 2009; Wei et al., 2009), ozonation (Qu et al., 2009), biodegra-
dation (Dirk et al., 2002; Kurosumi et al., 2008), photocatalysis
degradation (Muruganandham and Swaminathan, 2006) and
adsorption (Nigam et al., 2000; Kavitha and Namasivayam, 2007).
Some of these techniques decolorize wastewater by partially
decomposing the dye molecules and thereafter leaving the harmful
residues in the effluent. Alternatively, dye molecules, including
those that are too stable for the other methods, can be removed
completely by coagulation–flocculation. For this reason, the coagu-
* Corresponding author. Tel.: +86 591 83823131; fax: +86 591 22866539.
E-mail addresses: cxs2222@sina.com, chengxiansu@fzu.edu.cn (X. Cheng).
method to remove coloring agents from dye wastewater before
biological treatment.
Inorganic coagulants such as aluminum sulfate, ferric chloride
and polyaluminum chloride are widely used in many countries
(Lee et al., 2006; Joo et al., 2007). These inorganic coagulants are
inexpensive but have two major disadvantages: (a) they have to
be used in large quantities which result in a great volume of sludge
and (b) a high concentration of aluminum ion that remains in trea-
ted water may have adverse effect on human health. For these rea-
sons, cationic polymeric flocculants have received considerable
attention as an alternative for inorganic coagulants. Polymeric
flocculants can destabilize the coloring colloidal particles by com-
pressing their electrical double layers and subsequently forming
dye-polymer-dye bridges. Polyamine flocculants (Yue et al.,
2008), polydiallyldimethylammoniumchloride (PDADMAC) (Qi,
2007) and copolymer of acrylamide and 2-[(methacryloyl-
oxy)ethyl]trimethylammonium chloride [P(AM-DMC)] (Shen
et al., 2006) are all examples of cationic polymers that can be used
in dye wastewater treatment. Although a high level of dye removal
can be achieved by using these cationic polymeric flocculants,
industrial applications of them may be restricted because of the
high cost of the monomers. Therefore, an effective and economical
cationic flocculant, which is capable of dealing with the ever
increasing demand in this field, is desirable for industrial
application.
Lignin is the second most abundant natural organic resource on
earth, exceeded only by cellulose. The lignin used in this study was
isolated from the residue of enzymatically hydrolyzed cornstalks

0960-8524/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2010.04.094
7324 R. Fang et al. / Bioresource Technology 101 (2010) 7323–7329
as a by-product of bioethanol industry. Making use of this impor-
tant renewable biomass will not only benefit the bio-industry
but also diminish potential environmental pollutions. The aim of
this work was to (a) present a novel cationic copolymer which
can be synthesized by grafting the Mannich reaction product of
dimethylamine, acetone and formaldehyde onto hydroxymethylat-
ed lignin and (b) to evaluate the performance of this copolymer, as
a cationic flocculant, in removing anionic azo-dyes from aqueous
solutions.
This lignin-base dimethylamine–acetone–formaldehyde
copolymer (L-DAF) was tested for its capability to remove three an-
ionic dyes (Acid Black 1, Reactive Red 2 and Direct Red 23) from
simulated dye wastewaters via coagulation–flocculation. The
decolorization mechanism of L-DAF was discussed with respect
to its characteristics. The effect of solution pH, dye concentration
(C0), flocculant dose (m), settling time (t) and the addition of inor-
ganic salts on dye removal was studied to determine the optimum
operating conditions. Finally, the amount of sludge production and
the reduction of chemical oxygen demand (COD) were also
examined.
2. Methods
2.1. Materials
Lignin used in this study was isolated from the residue of enzy-
matically hydrolyzed cornstalks, and then purified in our labora-
tory. The raw material was supplied by Shandong Longlive Bio-
technology Co., Ltd., China. Dimethylamine (33 wt.% in aqueous
solution), formaldehyde (37 wt.% in aqueous solution), acetone,
HCl and NaOH were of analytical grade and purchased from Sinop-
harm Chemical Reagent Co., Ltd., China. Due to the unique isolation
procedures used, the density of functional groups, such as car-
bonyl, hydroxyl and phenolic hydroxyl groups, was very high in
the purified lignin. The weight-average molecular weight of lignin
is 2062 g/mol, the content of phenolic hydroxyl is 4.25 mmol/g
(Goldschmid, 1954), the content of residual sugar and ash is
0.22% and 0.39%, respectively.
Three commercially available anionic azo-dyes (Acid Black 1,
Reactive Red 2 and Direct Red 23) were purchased from Yuhua Eco-
nomic and Trade Corporation (Tianjin, China) and were used with-
out further purifications. Simulated dye wastewaters were
prepared by dissolving the appropriate amount of dye in 1 L of
deionized water. The pH of 100 mg/L of Acid Black, Reactive Red
and Direct Red liquor were 6.1, 6.5 and 6.5, respectively. Calibra-
tion curves were made for each dye at its absorbance maximum.
2.2. Preparation of L-DAF
The cationic flocculant used in this study, a lignin-base dimeth-
ylamine–acetone–formaldehyde copolymer (L-DAF), was synthe-
sized as follows: 20 ml dimethylamine solution, 9 ml acetone and
24 ml formaldehyde solution were added into a jacketed reactor
flask equipped with a stirrer and a reflux condenser. Then 2 mol/
L HCl was added dropwise in order to adjust the pH of the mixture
to a value of 3. A Mannich reaction was subsequently performed at
20 °C for 2 h. After completing the Mannich reaction, 5 g of lignin
was hydroxymethylated with formaldehyde in alkaline solution
for 10 min and thereafter fed into the reactor. At which a conden-
sation reaction was then carried out at 60 °C for 3 h. This procedure
gave a red–brown solution (L-DAF aqueous solution), the pH of this
L-DAF solution was between 8 and 9 and the solid content was
approximately 25%. The product was neutralized to pH 7 and puri-
fied by dialysis, vacuum dried to a constant weight and preserved
in a desiccator at room temperature.
2.3. Characterization of L-DAF
The cationicity of L-DAF was measured by chemical titration
whereby 0.5 g L-DAF powder was dissolved using 100 mL of deion-
ized water and 10 mL of sodium acetate/acetic acid buffer was
added to maintain a constant pH. Subsequently, 50 mL of potas-
sium ferricyanide (0.05 mol/L) was added into the solution and
the mixture was further diluted with deionized water and made
up to a total volume of 250 mL. The solution was left standing
for 1 h and afterwards filtered. Then 10 mL potassium iodide solu-
tion (0.6 mol/L), 10 mL HCl solution (6 mol/L) and 10 mL zinc sul-
fate solution (0.6 mol/L) were added to 100 mL of the filtrate.
Finally, titrated the free iodine with sodium thiosulfate solution
(0.1 mol/L), starch solution was used as an indicator. The cationi-
city was then calculated according to Eq. (1):
3  ðb  aÞ  N  2:5
C¼ ð1Þ
w
where C is the cationicity of L-DAF (mmol/g), a and b are the
volumes (mL) of sodium thiosulfate solution used in L-DAF titra-
tion and blank titration, N is the concentration of sodium thiosul-
fate solution (mol/L), w is the weight of L-DAF (g). The factor of 3
accounts for that 1 mol potassium ferricyanide can precipitate
3 mol of quaternary ammonium.
The molecular weight of L-DAF was determined with a Waters
515 GPC analysis instrument (Waters, US) using an Ultrahydrogel
250 column and 0.1 mol/L NaCl solution was used as the mobile
phase. A FT-IR2000 spectrometer (Perkinelmer, US) was employed
in the analysis of functional groups present on L-DAF. Purified L-
DAF samples were ground and pressed together with KBr to form
transparent films for FT-IR investigation. The zeta potential (f-po-
tential, an abbreviation for electrokinetic potential in colloidal sys-
tems) of flocs was measured using a JS94H zeta potential meter
(Zhongchen, China). After adding the appropriate amount of floccu-
lant into simulated dye wastewater, the mixture was stirred for
3 min. Then the sample was analyzed and the zeta potential was
recorded.
2.4. Coagulation–flocculation experiments
The coagulation–flocculation experiments with three anionic
dyes were conducted at room temperature using the HJ-2B jar-test
apparatus (Jinnan, China). These experiments consisted of three
steps: rapid mixing, slow mixing and sedimentation. Simulated
dye wastewater (100 mL) was transferred into a beaker and the
pH was adjusted to a desired value using HCl or NaOH solutions.
A certain amount of L-DAF was dissolved in deionized water and
added to the dye solution. The mixture was first stirred rapidly
at 200 rpm for 1 min and then at 40 rpm for 10 min. The suspen-
sion was left to settle for a period of time, then filtered by gravity
filtration and the filtrate was analyzed for dye concentration. It
was observed that the amount of the dye retained on filter paper
was negligible (less than 1%). In environmental chemistry, the
chemical oxygen demand (COD) test is commonly used to indi-
rectly measure the amount of organic pollutants found in the
water. It is expressed in milligrams per liter (mg/L), which indi-
cates the mass of oxygen consumed per liter of solution. The
COD of dye wastewater before and after treatment was measured
using AQ4001 COD tester (Orion, US) according to Chinese national
standard GB 11914-89.
To avoid the shift of kmax during the treatment, the pH of treated
dye solutions were brought to their initial state after coagulation–
flocculation. The dye concentration was determined by measuring
the absorbance at the kmax of each dye using a UV-2450 spectro-
photometer (Shimadzu, Japan) and the percentage removal of
dye was calculated according to the mass balance equation:
 R. Fang et al. / Bioresource Technology 101 (2010) 7323–7329 7325

ðC 0  CÞ Zeta potential is widely used in investigating the mechanism of

Dye removal ð%Þ ¼  100
C0
where C0 and C are the dye concentrations before and after treat-
ment (mg/L).
2.5. Statistical analyses
Each data point was taken as the average of three measure-
ments with standard deviation (STDEV). The experimental data
were analyzed statistically using Origin software (version 7.5). Lin-
ear regression analysis was used to describe the relationship be-
tween optimum L-DAF dose and dye concentration; one-way
analysis of variance (one-way ANOVA) was used to test the poten-
tial difference between treatments and dyes.
ð2Þ
coagulation and flocculation as it provides information regarding
the stability of colloidal dispersions. The zeta potential of flocs as a
function of L-DAF dose was tested. Meanwhile, the effect of dye con-
centration and flocculant dose on the removal of dyes was also inves-
tigated (Fig. 1). The aim of this comprehensive study is to further
understand the importance of charge neutralization in the dye re-
moval process.
All of the anionic dyes used in this study contain sulfonate
groups, which are negatively charged when dissolved in water.
(a) 100

90
3. Results and discussion

dye removal (%)

80
3.1. Approach of coagulation–flocculation mechanism

Acid Black 1
Functional groups in L-DAF were characterized by FT-IR analy- 70
sis. Upon the comparison between the spectrum of L-DAF and that C 0=250 mg/L

of the hydroxymethylated lignin, it was found that the relative C 0=150 mg/L
60
intensity of the adsorption bands at 3200–3600 cm1 (assigned C 0=100 mg/L

to O–H and N–H stretching vibration), 1608 cm1 (assigned to N– C 0=50 mg/L
50
H bending vibration) and 1480 cm1 (assigned to C–N bending
vibration of quaternary ammonium) increased significantly after
0 20 40 60 80 100 120 140 160 180
modification (Zhao et al., 2000). The appearance of a new peak at
dose of flocculant (mg/L)
about 1710 cm1 was assigned to the C@O stretching vibration of
carbonyl. These variations demonstrated that the condensation
compound of dimethylamine, acetone and formaldehyde was suc- (b) 100
cessfully grafted onto the lignin. The spectrum also showed that
the content of phenolic hydroxyl and carbonyl groups was very 90
high in L-DAF. This feature is very common in many bioflocculants
produced by micro-organisms, plants and animals. Mishra and Baj- 80
dye removal (%)

pai (2006) and Li et al. (2008) studied the flocculation perfor-

70
mances of Tamarindus mucilage and exopolysaccharide secreted
by bacterium, respectively. Both of them drew a conclusion that
60 Reactive red 2
functional groups play an important role in the flocculation behav-
C 0=250 mg/L
ior of these bioflocculants. Similarly, the functional groups con- 50
tained by L-DAF may strengthen the intermolecular interaction C 0=150 mg/L
C 0=100 mg/L
between the flocculant and the dye and enhance the bridging 40
C 0=50 mg/L
effect.
It is well known that both the cationicity and the molecular 30
weight of a cationic flocculant will significantly affect its coagula- 0 20 40 60 80 100 120
tion–flocculation performance (Guibal and Roussy, 2007). To reveal dose of flocculant (mg/L)
the effect of cationicity and molecular weight of L-DAF on the re-
moval of anionic dyes, three L-DAF samples (i.e. L-DAF 1, 2 and (c) 100
3) were prepared under varying reaction conditions. The cationi-
city of L-DAF 1, 2 and 3 was 1.79, 2.11 and 2.55 mmol/g and the
weight-average molecular weight was 2669, 2762 and 6143 g/ 90
dye removal (%)

mol, respectively. Their decolorization performances in treating

three anionic dyes (i.e. Acid Black 1, Reactive Red 2 and Direct 80
Red 23) were evaluated using one-way analysis of variance.
It was found that among these samples, L-DAF 3 (with the high- 70
est cationicity and molecular weight) was strongly superior to the Direct red 23
others in removing all three dyes from wastewater (P < 0.01), indi- C 0=250 mg/L
60 C 0=150 mg/L
cating that charge neutralization and bridging effect were probably
both responsible for the coagulation–flocculation of the dyes. C 0=100 mg/L
Although the molecular weight of L-DAF 2 was close to that of L- 50 C 0=50 mg/L
DAF 1, the percentage removal of all the dyes by L-DAF 2 was also
significantly higher than that of L-DAF 1 (P < 0.05). This may sug- 0 20 40 60 80 100 120
gest that charge neutralization plays a more important role than dose of flocculant (mg/L)
bridging in the removal of anionic dyes by L-DAF. As L-DAF 3 Fig. 1. Effect of dye concentration and flocculant dose on the removal of three
was the most effective product in decolorizing simulated dye anionic azo-dyes, (a) Acid Black 1; (b) Reactive Red 2; (c) Direct Red 23. pH 6.5,
wastewater, it was chosen for subsequent studies. T = 25 °C, t = 2 h.
7326 R. Fang et al. / Bioresource Technology 101 (2010) 7323–7329

Experimental results showed that the zeta potential of the flocs 160
changed from negative to positive with increasing L-DAF dose.
Moreover, when the initial zeta potential was reversed, maximum 140
color removal was achieved. This is because highly negative or po-

optimum L-DAF dose (mg/L)

120
sitive zeta potentials will confer stability to the dye molecules sus-
pended in wastewater, and the dispersion will resist aggregation. 100
However, when the zeta potential approaches the isoelectric point,
the negative charges carried by the dye molecules are neutralized 80
by L-DAF, the attractive forces between the dye molecules will ex-
ceed that of the repulsive forces and the dye dispersion will break 60
and flocculate. Therefore, the percentage removal of anionic dyes
40 Direct Red 23
will rise continuously with increasing L-DAF dose until the opti-
mum flocculant dose is reached, and when the optimum dose is ex- Recative Red 2
20
ceeded, a re-stabilization of the suspension takes place by the Acid Black 1
electrostatic repulsion among the positively charged particles. This 0
results in a less effective dye removal. 0 50 100 150 200 250 300
Although the chemical structure of this lignin derivative is not dye concentration C0 (mg/L)
well established, the presence of many functional groups, espe-
cially cationic groups, on L-DAF has been confirmed by IR analysis, Fig. 2. Relationship between optimum L-DAF dose and dye concentration.
chemical titrations and measurements of zeta potentials. Further-
more, the mechanism for the removal of anionic dyes by coagula-
tion–flocculation using L-DAF can be ascribed to two simultaneous Table 1
mechanisms, which are coagulation by charge neutralization and Linear relationship between optimum L-DAF dose and dye concentration C0.
flocculation by bridging. Some bio-polymers have large molecular Dyes A B R
size and high flocculating activity on various suspended solids
Acid Black 1 13.00 ± 7.08 0.56 ± 0.04 0.9935
(Chakrabarti et al., 2008; Li et al., 2008). In these cases, bridging ef- Reactive Red 2 13.71 ± 1.55 0.35 ± 0.01 0.9992
fect is found to be the major factor. However, when a low molecu- Direct Red 23 5.57 ± 5.12 0.43 ± 0.03 0.9943
lar weight cationic polymer is used, charge neutralization becomes
Optimum L-DAF dose = A + B  C0.
the dominant mechanism involved in the coagulation–flocculation A: Intercept value and its standard error; B: slope value and its standard error; R:
process (Yan et al., 2004). L-DAF has a relatively low molecular correlation coefficient.
weight (103–104 g/mol) when compared with other commonly
used flocculants, such as polyacrylamide, polysaccharide and
amount of anionic particles and thus more cationic flocculants are
chitosan derivatives (104–106 g/mol). Thus, as proposed by Yan
required to complete the charge neutralization step. Both phenom-
et al. (2004), our results also showed that with L-DAF as the cat-
enon typically correspond to a coagulation–flocculation system
ionic flocculant, charge neutralization is the main mechanism in-
controlled by charge neutralization mechanism (Szygula et al.,
volved in the coagulation–flocculation process.
2008), which supports the explanation proposed in Section 3.1.

3.2. Coagulation–flocculation studies

In order to optimize the removal of dyes using L-DAF which in-
volves more than one mechanism, it is important to consider the
factors that may have an effect on the mechanisms of this process.
In this study, the effect of some of these factors, for example, dye
concentration (C0), dose of flocculant (m), solution pH, settling time
(t) and the presence of inorganic salt on the decolorization effi-
ciency of L-DAF was investigated.
3.2.1. Effect of dye concentration (C0) and flocculant dose (m)
Fig. 1 shows the effect of dye concentration and flocculant dose
on the removal of three anionic dyes. The dye concentration was
varied so that the optimum dose of L-DAF for each dye can be
determined.
The results shown in Fig. 1 suggested that for all three dyes, the
efficiency of dye removal first increased and then decreased with
increasing dose of L-DAF. There was an optimum L-DAF dose for
each simulated wastewater at which almost complete dye removal
could be achieved. Furthermore, when the dye concentration was
increased from 50 mg/L to 250 mg/L, a higher dose of L-DAF was
required to achieve a satisfactory level of color removal. The linear
relationship between optimum L-DAF dose and dye concentration
was revealed using linear regression analysis (Fig. 2):
Optimum L-DAF dose ¼ A þ B  C 0
Parameters A, B and the correlation coefficients (R) are listed in
Table 1. The relationship is significant for all the dyes at the 0.01
level, indicating that at a higher dye concentration, there is a larger
3.2.2. Effect of solution pH
The removal of anionic azo-dyes is directly affected by the
strength and the availability of positive charges, which is in turn
dependent on the cationicity and the conformation of the L-DAF
dissolved in aqueous solution. Furthermore, the conformation of
the dissolved L-DAF (as a polyelectrolyte) is influenced by the pH
of the solution. This theory is supported by Somasundaran and
Runkana (2005) who concluded that the pH of the dye solution
influences the electrochemical nature of the particles as well as
the dissociation of the polyelectrolytes and hence their conforma-
tion in solution. As a result, the effect of pH on the removal of dyes
was investigated and the results are shown in Fig. 3.
In Fig. 3, the optimum dose of L-DAF required for a complete
dye removal decreased with decreasing pH. This phenomenon sug-
gests that the decolorization efficiency of L-DAF was improved
when the dye solution shifted from being basic to being acidic.
Similar trend can be observed in the case of deacetylated chitosan,
which is a representative bioflocculant widely used in wastewater
treatment. At a pH level lower than 5.0, more than 90% of the
amine groups contained by chitosan will be protonated (Szygula
et al., 2009). Therefore, a lower pH increases the cationic strength
of chitosan and then improves its flocculation ability. However, L-
DAF is a little different in molecular structure. It is a highly posi-
tively charged polyelectrolyte with relatively small molecular size.
ð3Þ
Under acidic conditions, L-DAF exists in a stretched form which en-
hanced the availability of the positive charges and the bridging
capability of the flocculant (Somasundaran and Runkana, 2005).
Alternatively, as the solution pH increases from 4.0 to 9.0, the phe-
 R. Fang et al. / Bioresource Technology 101 (2010) 7323–7329 7327

moval was investigated. L-DAF (5 mg) was dissolved in deionized

(a) 100
water and added to a dye solution (100 mL) in a 250 mL beaker.
90 After agitation and settling for a period of time, a sample of the
supernatant was taken from the dye solution, at a level of 1 cm be-
80 low the surface, and the residual dye concentration of the sample
dye removal (%)

70
was measured.
Fig. 4 shows the effect of settling time on the percentage of dye
60 removal. During the experiments, large quantities of small flocs
could be observed just after rapid agitation. The small flocs contin-
50
ued to aggregate to form larger ones and settled at the bottom of
40 Acid black 1 the beaker. It can also be concluded from Fig. 4 that the coagula-
pH=4.0 tion–flocculation process was fast with most of the flocs settled
30
pH=6.5 within 2 h. The rapid sedimentation indicated that the process is
20 pH=9.0 controlled by both charge neutralization and bridging (Szygula
et al., 2008; Beltran-Heredia et al., 2009). The experiments were
20 30 40 50 60 70 80 90 100 110 carried out for a total of 4 h but the residue dye concentration re-
dose of flocculant (mg/L) mained fairly constant after 2 h. Therefore, 2 h is considered to be
the suitable settling time. Furthermore, the most recent experi-
(b) 100 ments with 1 mg/L of anionic polyacrylamide (HPAM) as a coagu-
lant aid revealed that the settling speed could be further enhanced
90 and a complete settlement could be achieved within 30–40 min.

80
dye removal (%)

70
3.2.4. Effect of inorganic salt
60 Various types of inorganic salts can also be found in textile
effluent. This gives rise to high ionic strengths which may influence
50 Reactive red 2 the decolorization efficiency of the flocculants. For this reason, the
pH=4.0 effect of chloride, nitrate and sulfate ions on the removal of dyes
40
pH=6.5 was investigated. In these experiments, the salt concentration
30 pH=9.0 was 0.05 mol/L, the dye concentration was 100 mg/L and the dose
of L-DAF was 75, 50 and 35 mg/L for Acid Black, Reactive Red and
20 Direct Red solution, respectively. In addition, one experiment was
20 30 40 50 60 70 80 90 100 110 carried out using tap water instead of deionized water.
dose of flocculant (mg/L) It was found that the addition of some inorganic salts, such as
NaCl and NaNO3, and the replacement of deionized water by tap
(c) 100 water had no significant (P > 0.05) effect on the decolorization effi-
ciency of L-DAF. The dye removal of all three anionic dyes re-
90 mained fairly constant at a high salt concentration. The only
80 exception was with the addition of Na2SO4, which led to a decline
dye removal (%)

of 2–4% in dye removal. This study suggests that the cation Na+ and
70 the anions like Cl and NO 3 as well as other ions contained in tap

60
water do not react with L-DAF during the coagulation–flocculation
process. The slight decrease of dye removal in the presence of high
50
Direct red 23
40
pH=4.0
30 100
pH=6.5
pH=9.0
20 90
20 30 40 50 60 70 80 90 100 110
dose of flocculant (mg/L) 80
dye removal (%)

Fig. 3. Effect of solution pH on the removal of three anionic azo-dyes, (a) Acid Black 70
1; (b) Reactive Red 2; (c) Direct Red 23. C0 = 100 mg/L, T = 25 °C, t = 2 h.

60
nolic group of lignin turns to phenolate anion and the conforma-
tion of the polymer changes from stretched to coiled form, both
50
of which lead to the decline of the decolorization efficiency of L-
Direct Red 23
DAF. Thus, lower dose of flocculant was required for acidic condi-
40 Reactive Red 2
tions than that of basic conditions. Acid Black 1
30
3.2.3. Effect of settling time (t)
0 30 60 90 120 150 180 210 240
In the coagulation–flocculation process, the settling speed of
settling time (min)
the formed flocs is of great importance as this parameter influences
the design of the settling tank (Szygula et al., 2008). To evaluate the Fig. 4. Effect of settling time on the removal of three anionic azo-dyes. C0 = 100 mg/
settling speed of the flocs, the effect of settling time on the dye re- L, m = 50 mg/L, pH 6.5, T = 25 °C.
7328 R. Fang et al. / Bioresource Technology 101 (2010) 7323–7329
Table 2
Sludge production and the COD reduction of dye wastewaters (C0 = 100 mg/L, pH 6.5, T = 25 °C, t = 24 h)a.
Dye Dose of L-DAF (mg/L) Dye removal (%) Sludge production (%)
Acid Black 1 75 97.1 ± 0.5 5.3 ± 0.4
Reactive Red 2 50 98.3 ± 0.4 5.0 ± 0.2
Direct Red 23 35 99.5 ± 0.2 4.0 ± 0.2
a
Data are expressed as mean ± standard deviation (STDEV) of triplicate measurements.
concentration of SO2
4 may be due to a weak interaction between L-
DAF and the sulfate ions.
3.3. Sludge production and the removal of COD
Besides color removal, the amount of sludge production and the
reduction of COD are also important factors for practical applica-
tion of wastewater treatment and therefore need to be determined.
The amount of sludge production is greatly affected by the dose
of flocculant and the operating conditions. The volume of sludge
produced during the treatment of dye wastewaters by L-DAF was
measured using an Imhoff cone (Szygula et al., 2009). These exper-
iments were carried out by adding the optimum dose of L-DAF into
300 mL various dye solutions (100 mg/L). Once the samples were
mixed thoroughly, the contents in the beakers were transferred
to an Imhoff cone and the volume of sludge was determined. De-
spite previous findings that most flocs settled within 2 h, settling
time was increased to 24 h to ensure complete floc settlement
and sludge production. The volume of sludge production is listed
in Table 2. When compared with polyaluminum chloride (PAC),
which is widely used in practice as a cationic coagulant in China
and has a sludge production of 8–15%, the volume of the sludge
produced during the treatment using L-DAF is much lower; 5.3%
sludge production for Acid Black, 5.0% for Reactive Red and 4.0%
for Direct Red.
Some treatment methods, such as biological and chemical deg-
radation, offer excellent means for the decolorization of dye waste-
water, but they are relatively inefficient in reducing the COD of the
effluent (Kurosumi et al., 2008; Qu et al., 2009). In this study, the
COD values of dye wastewaters before and after treatment using
L-DAF were determined and presented in Table 2. It is apparent
that, in addition to color removal, L-DAF is efficient in reducing
the COD of dye wastewater. Under specific conditions, the COD
reduction was 89.3%, 96.3% and 93.5% for Acid Black, Reactive
Red and Direct Red, respectively. The COD values of these treated
dye wastewater range from 11.8 mg/L to 30.2 mg/L. All of which
are within the limit set by Chinese environmental regulation GB
4287–92 (i.e. COD <100 mg/L for textile industry).
4. Conclusions
This study demonstrates that L-DAF is an efficacious cationic
flocculant for the removal of anionic azo-dyes from aqueous solu-
tion. The removal of dyes by L-DAF can be attributed to charge neu-
tralization and bridging effect, among which charge neutralization
is the dominant mechanism. The decolorization efficiency of L-DAF
can be enhanced by decreasing solution pH and is not affected by a
high salt concentration. There is an optimum flocculant dose for
each dye wastewater and dye removal close to 100% can be
achieved under suitable conditions. The COD removal and the
sludge production can meet the government’s requirement for
commercial applications.
COD before treatment (mg/L) COD after treatment (mg/L) COD reduction (%)
281.7 30.2 ± 1.7 89.3 ± 0.8
315.6 11.8 ± 1.3 96.3 ± 0.5
283.8 18.5 ± 1.8 93.5 ± 0.8
Acknowledgements
The authors would like to thank the Patent Transfer Fund of
Development and Reform Committee of Fujian Province (2008-
08) for their financial support. Also, we are especially grateful to
Mr. Ocean Cheung of Stockholm University, Sweden, for his contri-
bution towards this paper.
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