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 Process Biochemistry 40 (2005) 2545–2550
www.elsevier.com/locate/procbio

Kinetic analysis of dyestuff and COD removal from synthetic

wastewater in an anaerobic packed column reactor
Ilgi Karapinar Kapdan*
Department of Environmental Engineering, Dokuz Eylül University, Buca Tinaztepe Campus, Izmir, Turkey
Received 15 April 2004; received in revised form 7 October 2004; accepted 22 November 2004

Abstract

An anaerobic packed column reactor was operated continuously at different dyestuff loading rates, (0.05–0.4 g/(l day)) and COD loading
rates (1–8 g/(l day)) in order to determine dyestuff and COD removal kinetic constants. The system was operated at room temperature
(20
1 8C) and at pH 7 using an immobilized anaerobic bacterial consortium called PDW. Synthetic wastewater containing textile dyestuff
Reactive Red 195 was fed from the bottom of the reactor. Around 90% decolourization efficiency was obtained for dyestuff loading rates up to
0.15 g/(l day). COD removal efficiency was obtained between 5 and 35% for the applied loads. Modified Stover–Kincannon model was
applied to the experimental data. Saturation value constant and maximum utilization rate constant of Stover–Kincannon model for dyestuff
and COD were determined as KB = 17.8 g/(l day), Umax = 19.5 g/(l day) and KB = 37.9 g COD/(l day), Umax = 12.9 g COD/(l day),
respectively. The predicted effluent dyestuff and COD concentrations were calculated using the constants and it was found that they are
in good agreement with the observed ones.
# 2004 Elsevier Ltd. All rights reserved.

Keywords: Anaerobic treatment; COD; Decolourization; Kinetic analysis; Stover–Kincannon model

1. Introduction anaerobic conditions and high decolourization efficiencies

Colour removal under anaerobic condition could be
biodegradation of dyestuff by azoreductase activity [1,2] or
nonenzymic azo reduction in dyestuff [3–5]. However, the
azoreductase cleavage of azo bond may result in formation
of aromatic amines which might be toxic [3,6]. But, some
reports showed that the effluent of anaerobic decolourization
process were completely non-toxic [5,7].
Studies indicated that colour removal under anaerobic
conditions is significantly affected by dyestuff structure.
Azo type dyestuffs are readily biodegradable while metal
complex, antraquanin and indigo group dyestuffs are not
[8,9]. In addition, dyestuff can be used as sole carbon
sources by the microrganisms in some cases but co-substrate
as carbon source addition is required [10–12]. Decolouriza-
tion of textile dyestuffs have been carried out in UASB,
sequencing batch, fed-batch and packed bed operated under
* Fax: +90 232 4531143.
E-mail address: ilgi.karapinar@deu.edu.tr.
were obtained [13–18]. It was also reported that an aerobic
unit after anaerobic decolourization unit act as a polishing
step, provides higher pollutant removal, like COD and toxic
substances, rather than decolourization [19,20].
Determination of kinetic constants of a bioprocess is a
useful tool to be able to describe and to predict the
performance of the system. In this study, an anaerobic
packed column reactor was operated continuously at
different dyestuff and COD loading rates in order to
evaluate the decolourization performance and to determine
kinetic constants.
2. Mathematical model
Monod type kinetic analysis based on substrate, cell mass
and dyestuff concentration in batch operation for decolour-
ization purpose have been used [8,10]. Anaerobic decolour-
ization and batch inhibition kinetics of different azo dyes
have also been investigated [21,22]. In addition, there are

0032-9592/$ – see front matter # 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.procbio.2004.11.002
2546 I.K. Kapdan / Process Biochemistry 40 (2005) 2545–2550
kinetic models developed for organic substance removal in
continuously operated anaerobic reactors [23–26]. Stover–
Kincannon is one of the most widely used mathematical
model for determining the kinetic constants in immobilized
systems. The model have been applied to continuously
operated mesophilic and thermophilic upflow anaerobic
filters for treatment of paper-pulp liquors [27] and simulated
starch wastewater [28], anaerobic filter for soybean waste-
water treatment [29], anaerobic hybrid reactor [30], aerobic
treatment of synthetic dairy wastewater in trickling filters
[31] and municipal wastewater treatment in submerged
biofilters [32]. However, this model has not been applied for
determination of decolourization kinetic constants. There-
fore, the Stover–Kincannon model was used for the kinetic
analysis of COD and dyestuff removal in an upflow
anaerobic packed bed reactor in this study.
The Stover–Kincannon model considers the organic
substance removal rate as a function of organic loading rate
at steady state as in Eq. (1).
dS Q
¼ ðS0  Se Þ (1)
dt V
The original Stover–Kincannon model for rotating biofilm
reactor is described as in Eq. (2).
dS QðS0  Se Þ Umax ðQS0 =AÞ
¼ ¼ (2)
dt V KB þ ðQS0 =AÞ
where A represents the total disc surface area whereby total
biomass concentration immobilized on discs. The suspended
biomass concentration is assumed to be negligible compare
to that of attached biomass. The simple modification of the
original Stover–Kincannon model is the introduction of total
organic loading rate, QS0/V into the Eq. (1) instead of QS0/A,
resulting in Eq. (3). Since void space has a significant
importance in obtaining high removal efficiency in anaero-
bic filters the modified Stover–Kincannon model has been
applied to anaerobic filters and the fixed bed region of hybrid
reactors [29,30].
dS QðS0  Se Þ Umax ðQS0 =VÞ
¼ ¼ (3)
dt V KB þ ðQS0 =VÞ
linearization of Eq. (3) gives the relationship:
dS V KB V 1
¼ ¼  þ (4)
dt QðS0  Se Þ Umax QS0 Umax
where dS/dt is the substrate removal rate (g/(l day)), S the
substrate concentration in the reactor (g/(l day)), Umax the
maximum substrate removal rate constant (g/(l day)) and KB
is a saturation value constant (g/(l day)). The plot of V/
[Q(S0  Se)], inverse of the loading removal rate, versus V/
(QS0), inverse of the total loading rate will result in a straight
line. The intercept and slope of the line results in 1/Umax and
KB/Umax, respectively.
The substrate balance for the reactor can be written as
follows:
 
dS
QS0 ¼ QSe þ V (5)
dt
substitution of Eq. (3–5) results in
Umax ðQS0 =VÞ
QS0 ¼ QSe þ V (6)
KB þ ðQS0 =VÞ
This expression can then be solved for either the effluent
substrate concentration (Eq. (7)) or the required volume of
the anaerobic filter (Eq. (8)) by substituting kinetic constants
Umax and KB.
Umax S0
Se ¼ S0  (7)
KB þ ðQS0 =VÞ
QS0
V¼ (8)
ðUmax S0 =S0  Se Þ  KB
3. Material and methods
3.1. Microbial culture
A facultatively anaerobic bacterial culture, isolated by
the Biotechnology Centre at the University of Ulster, N.
Ireland [33] was used in decolourization of dyestuff. The
facultative anaerobic bacterial consortium called PDW
consists of Alcaligenes faecolis and Commomonas acid-
ourans. The culture was grown in flasks at its optimum
growth temperature 28 8C and preserved at 4 8C.
3.2. Media composition and dyestuff
Nutrient media used for PDW culture had the following
composition: 0.5 g/l (NH4)2SO4; 2.66 g/l KH2PO4; 4.32 g/l
Na2HPO42H2O and dyestuff. Molasses was used as carbon
source. The textile dyestuff used was Reactive Red 195
monoazo based, viniysulphone/monochlorotiazine bifunc-
tional and obtained from EKOTEN Co., Izmir, Turkey in
pure form.
3.3. Experimental set-up
The anaerobic packed column reactor was made up of
chrome–nickel with the reactor liquid volume of 6000 ml.
The support particles used for the immobilization of
microorganisms were made of mesh of stainless steel metal
wire. The total surface area of support material was
192.56 m2. The system was fed from the bottom with
dyestuff containing synthetic wastewater by a peristaltic
pump and effluent was collected from the top.
At the beginning of the experiments, the anaerobic unit
was inoculated with dene PDW culture (3 g/l) cultivated in
flasks and was operated batch-wise for 2 weeks to
immobilize the culture on wire meshes. The system was
then operated continuously for 1 month to increase the
biomass concentration in the system and to reach stable
anaerobic conditions at start-up phase. Each experimental
study was carried out at least two weeks until the system
 I.K. Kapdan / Process Biochemistry 40 (2005) 2545–2550
reached steady state conditions. Nitrogen gas was passed
through the reactor to make the conditions anaerobic.
3.4. Analytical methods
Daily samples were withdrawn and centrifuged at
5000 rpm until a clear supernatant was obtained. Analyses
were carried out on clear supernatants.
A scanning spectrophotometer (Novaspec II, Pharmacia
Biotech.) was used for colour measurements. Absorbance
measurements were done at the maximum absorbance
wavelength of the dyestuff Reactive Red 195 (l = 520 nm).
Samples were diluted with distilled water prior to
measurements if necessary. Absorbance values and dyestuff
concentration were evaluated from the developed absor-
bance–concentration curve. Dyestuff concentration was
used in calculation of decolourization efficiency. COD
analyses were carried out on clear supernatant according to
standard methods [34].
4. Results and discussions
4.1. Effect of dyestuff loading rate on dyestuff removal
The variation of effluent dyestuff concentration and
decolourization efficiency with dyestuff loading rate is given
in Fig. 1. The anaerobic unit operated with high
decolourization performance between 0.05 and 0.175 g/
(l day) dyestuff loading rates (DLR) with over 90%
decolourization efficiency which results in effluent dyestuff
concentration lower than De = 0.015 g/l. For higher DLR
(>0.3 g/(l day)), the efficiency sharply decreased to 50%
with De = 0.150 g/l. These results indicated that anaerobic
packed bed column can be operated over 90% decolouriza-
tion efficiency up to 0.2 g/(l day) dyestuff loading rates. The
maximum dyestuff removal rate (r = (Q  (D0  De)/V)
Fig. 1. Variation of effluent dyestuff concentration and dyestuff removal
efficiency with dyestuff loading rate. (*) Effluent dyestuff concentration;
(*) efficiency.
2547
Fig. 2. Variation of dyestuff removal rate with effluent dyestuff concentra-
tion.
was obtained as 0.217 g/(l day) with an effluent dyestuff
concentration of 0.173 g/l (Fig. 2).
4.2. Effect of organic loading rate on COD removal
Fig. 3 depicts the variation of effluent COD concentration
and COD removal efficiency with COD loading rate
(Q  S0/V). Effluent COD concentration was around 2 g/l
for organic loading rate between OLR = 2.5–4 g/(l day) and
then increased to Se = 4 g/l for OLR = 5 g/(l day). For the
high OLR (>7 g/(l day)), effluent COD concentration was
increased to Se = 7.3 g/l. COD removal efficiency was
around 35 % for OLR up to 5 g/(l day) and decreased to 10%
for OLR = 8 g/(l day). Almost, no COD removal was
obtained at high organic loading rates.
The variation of COD removal rate with effluent COD
concentration is depicted in Fig. 4. COD removal rate
increases from 0.46 to 1.4 g/(l day) by reaching the
maximum level for the effluent COD concentration between
0.9 and 3 g/l, respectively and decreases to 0.68 g/(l day) as
effluent COD concentration increases to 7.32 g/l. Low COD
removal rates are obtained in anaerobic packed column
reactor during the decolourization process. This could be
Fig. 3. Variation of effluent COD concentration and COD removal effi-
ciency with organic loading rate. (*) Effluent COD concentration; (*)
efficiency.
2548 I.K. Kapdan / Process Biochemistry 40 (2005) 2545–2550
Fig. 4. Variation of COD removal rate with effluent COD concentration.
because of operational conditions including temperature
(20 8C) and synthetic media composition which was
adjusted to obtain colour removal rather than COD removal
and do not enhance the growth of methanogenic bacteria
which are the main agents in COD removal in the anaerobic
process. In addition, the presence of dyestuff or biodegrada-
tion end products in the reactor might have had toxic or
inhibitory effects on the microbial culture that resulted in
low COD removal. In order to improve COD removal, the
system can be operated at longer hydraulic retention times or
an aerobic unit after anaerobic unit can be used as a
polishing step.
4.3. Determination of modified Stover–Kincannon model
constants for dyestuff and COD
The modified Stover–Kincannon model was applied to
experimental results from the continuously operated up flow
anaerobic packed bed reactor for decolourization of textile
dyestuff and kinetic constants for dye removal and COD
removal were determined. Dyestuff loading rates and
removal rates were calculated at different hydraulic
retention times and initial dyestuff concentrations. Fig. 5
indicates the plot of dyestuff loading V/(QD0) versus V/
[Q(D0  De)] dyestuff removal rate. From the slope and
intercept of a best-fit line (R2 = 0.99), kinetic constants for
Fig. 5. Stover–Kincannon model plot for dyestuff removal.
Fig. 6. Relationship between observed and predicted effluent dyestuff
concentrations.
dyestuff removal were determined as Umax = 0.47 g/(l day)
and KB = 0.43 g/(l day), respectively.
Therefore the rate expression for dyestuff takes the
following form:
QðD0  De Þ 0:47ðQD0 =VÞ
¼ (9)
V 0:43 þ ðQD0 =VÞ
and effluent dyestuff concentration can be predicted by using
the Eq. (9).
0:47D0
De ¼ D0  (10)
0:43 þ ðQD0 =VÞ
Fig. 6 indicates the relationship between observed dyestuff
concentrations from the experiments carried out under
different operational conditions and predicted dyestuff con-
centrations calculated by using Eq. (10). There is a good
agreement between observed and predicted dyestuff con-
centrations with R2 = 0.97 regression coefficient.
Similarly, when the model is applied to COD removal
(Fig. 7), the coefficients are obtained as Umax = 12.99 g
COD/(l day), KB = 37.69 g COD/(l day) with high regres-
Fig. 7. Stover–Kincannon model plot for COD removal.
 I.K. Kapdan / Process Biochemistry 40 (2005) 2545–2550
Fig. 8. Relationship between observed and predicted effluent COD con-
centrations.
2
sion coefficient (R = 0.97). Rate expression for COD takes
the following forms,
QðS0  Se Þ 12:99ðQS0 =VÞ
¼ (11)
V 37:69 þ ðQS0 =VÞ
Fig. 8 indicates the relationship between predicted and
observed effluent COD concentration in anaerobic reactor.
Predicted effluent COD concentration was calculated by
using the following equation:
12:99S0
Se ¼ S0 
37:69 þ ðQS0 =VÞ
There is a linear relationship between observed and pre-
dicted effluent COD concentrations with R2 = 0.9501 regres-
sion coefficient indicting that kinetic constants can be used
in predicting effluent COD concentration of anaerobic
packed column reactor.
5. Conclusions
An upflow anaerobic packed bed reactor was operated
continuously at different dyestuff loading and COD loading
rates to remove the colour from synthetic wastewater. The
colour removal efficiency was around 90% up to 0.15 g/
(l day) dyestuff loading rate with effluent dyestuff con-
centration of 0.06 g/l. Dyestuff was removed almost at the
same rate up to 0.15 g/(l day) dyestuff loading rate means
that most of the dye was removed by microbial action.
Maximum dyestuff removal rate was 0.217 g/(l day) with
effluent dyes concentration of De = 0.17 g/l. COD removal
efficiency varied between 10 and 35% for the organic
loading rate between 1 and 8 g/(l day). The maximum COD
removal rate was obtained as 1.4 g/(l day) with the effluent
COD concentration of Se = 4 g/l at OLR = 5 g/(l day). The
experimental results indicated that high decolourization can
be obtained at anaerobic packed column operated at room
temperature (20
1 8C) with low COD removal. An aerobic
2549
unit can be used as polishing step to enhance the COD
removal or system can be operated at higher temperature to
increase the activity of methanogenic bacteria.
As a result of the kinetic analysis of reactor for dye and
COD removal using a modified Stover–Kincannon model,
the substrate utilization rate, Umax, and saturation constant
value, KB, was determined as Umax = 0.47 g/(l day) and
KB = 0.43 g/(l day) for dye and as Umax = 12.99 g COD/
(l day), KB = 37.69 g COD/(l day) for COD removal. There
was good agreement between observed and predicted
concentrations both for dyestuff and COD means that
kinetic constants can be used to design an anaerobic packed
column for colour and COD removal from textile effluents.
Acknowledgement
This study was supported by research funds of Dokuz
Eylül University, Izmir, Turkey and Scientific and Technical
Research Council of Turkey.
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