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COD Fractionation and Biodegradation Kinetics of Segregated Domestic

Wastewater: Black and Grey Water Fractions

Article  in  Journal of Chemical Technology & Biotechnology · September 2010

DOI: 10.1002/jctb.2423

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Research Article
Received: 27 January 2010 Revised: 26 March 2010 Accepted: 31 March 2010 Published online in Wiley Interscience: 3 June 2010

(www.interscience.wiley.com) DOI 10.1002/jctb.2423

COD fractionation and biodegradation kinetics

of segregated domestic wastewater: black
and grey water fractions
Selda Murat Hocaoglu,∗ Güçlü Insel, Emine Ubay Cokgor, Ahmet Baban
and Derin Orhon

Abstract
BACKGROUND: Household wastewater was segregated to grey and black water components as part of a recent approach aiming
to separate wastewater fractions for more efficient treatment and reuse. Chemical oxygen demand (COD) fractionation and
biodegradation characteristics of black and grey water were determined by means of model calibration of the oxygen uptake
rate profiles. An ASM1 type of model modified for endogenous respiration was selected for this purpose.

RESULTS: Calibration identified, aside from the readily biodegradable COD, two hydrolysable COD fractions in the black water,
and only a single, slowly biodegradable COD fraction, in the grey water. The relative magnitude of the readily biodegradable
COD fraction was high compared with typical domestic sewage, especially for grey water, and the level of particulate inert COD
fraction, was relatively low. Biodegradation of soluble COD fractions in both wastewaters approximated typical rates observed
for domestic sewage. However, the hydrolysis rate of the slowly biodegradable COD was quite low, reflecting the characteristics
and impact of settleable organic matter.

CONCLUSION: Based on the process rates obtained, aerobic biodegradation was equally effective for black and grey water
fractions, both for the removal of soluble COD and the stabilization of particulate COD.

c 2010 Society of Chemical Industry

Keywords: black water; grey water; COD fractions; process kinetics; respirometry; MBR

NOTATION VSS: Volatile suspended solids [mg L−1 ]

−1 VT : Total reactor volume [L]
bH : Endogenous decay rate for XH [day ]
BOD5 : Biological oxygen demand [mgO2 L−1 ] VWW : Initial wastewater volume [L]
COD: Chemical oxygen demand [mgO2 L−1 ] XI : Particulate inert COD [mgCOD L−1 ]
CS : Biodegradable COD [mgCOD L−1 ] XH : Active heterotrophic biomass [mgCOD L−1 ]
CT : Total COD [mgCOD L−1 ] Xo: Initial mass of biomass in respirometric tests [mg L−1 ]
OUR: Oxygen uptake rate [mg (L.h)−1 ] XS1 : Rapidly hydrolysable COD [mgCOD L−1 ]
kh : Maximum hydrolysis rate for XS1 [day−1 ] XS2 : Slowly hydrolysable COD [mgCOD L−1 ]
KX : Hydrolysis half saturation constant for XS1 [gCOD (g XT : Particulate COD [mgCOD L−1 ]
cellCOD)−1 ] YH : Heterotrophic Yield Coefficient [g cellCOD (gCOD)−1 ]
khx : Maximum hydrolysis rate for XS2 [day−1 ] YSP : Yield for soluble microbial products [−]
KS : Half saturation constant for growth of XH [mgCOD
L−1 ] INTRODUCTION
KXX : Hydrolysis half saturation constant for XS2 [gCOD (g Until recently, characterization surveys of wastewaters were
cellCOD)−1 ] limited to the assessment of a few parameters considered to be
µHmax : Maximum growth rate for XH [day−1 ] important for the treatment scheme to be adopted. In the conven-
MLVSS: Mixed liquor volatile suspended solids [mg L−1 ] tional approach, the organic carbon content of the wastewater
Sı: Soluble inert COD [mgCOD L−1 ] was characterized by collective parameters such as BOD5 , and
SP : Soluble microbial products [mgCOD L−1 ]
SS : Readily biodegradable COD [mgCOD L−1 ]
ST : Soluble COD [mgCOD L−1 ] ∗
Correspondence to: Selda Murat Hocaoglu, TUBITAK Marmara Research Center,
TKN: Total Kjeldahl Nitrogen [mgN L−1 ] Environment Institute, 41470, Gebze, Kocaeli, Turkey.
TN: Total nitrogen [mgN L−1 ] E-mail: selda.murat@mam.gov.tr
TP: Total phosphorus [mgP L−1 ]
1241

TUBITAK Marmara Research Center, Environment Institute, 41470, Gebze,

TSS: Total suspended solids [mg L−1 ] Kocaeli, Turkey

J Chem Technol Biotechnol 2010; 85: 1241–1249 www.soci.org

c 2010 Society of Chemical Industry
 www.soci.org
COD. Evaluations and even modeling attempts were performed,
although quite unsuccessfully, using these overall indicators.1
Adoption of COD as the main organic carbon parameter and
recognition of COD fractions with different biodegradation char-
acteristics have been major milestones for new developments.2
Research efforts (i) established the electron equivalence between
COD utilized, biomass generated and oxygen consumed in
biological treatment systems, and (ii) promoted dissolved oxygen
as an important model component as an additional parameter
for the evaluation of biodegradation characteristics.3,4 New multi-
component models were developed involving COD fractionation,
and experimental assessment associated with these models relied
on measuring changes in the oxygen demand with time, or more
precisely, on respirometric methods measuring the oxygen uptake
rate under a controlled environment.5 – 7 Methods using respirom-
etry have been quite successful in assessing COD fractions of
wastewaters,8,9 as well as associated kinetic and stoichiometric
model parameters.10 – 12 Modeling and calibration of appropriate
models using experimental data, and especially oxygen uptake
rate profiles, were also developed as an efficient approach to
evaluate biodegradation characteristics of wastewaters.13 – 15
These efforts enabled the establishment of a significant database
for different wastewaters.16 Obviously, this line of experimental
investigation has enhanced aerobic biological processes. A similar
approach is yet to be developed for anaerobic processes.
Characterization of biodegradation, while extensively used for
domestic sewage and different industrial wastewaters, has not
been used for fractions of domestic sewage, namely, black water
and grey water fractions. These fractions emerged as a result of a
recent approach that aims to segregate and collect sewage in two
main fractions for more efficient treatment and reuse.17,18 Black wa-
ter is usually restricted to toilet wastes, containing the majority of
nutrients, which may be utilized for agricultural purposes. Grey wa-
ter is the remaining wastewater originating from bathtubs, show-
ers, hand-washing basins, washing machines, and kitchens.18,19
Some studies have excluded the kitchen wastewater stream from
grey water.20,21 Grey water was generally considered to have lower
concentrations of organic compounds, and fewer pathogens than
that of combined domestic wastewater. In this context, it was
evaluated as a potential resource for recovery and reuse after
appropriate treatment.22 Conventionally, aerobic treatment was
found suitable for grey water to give satisfactory effluent quality,23
whereas black water was generally treated by anaerobic treatment
systems, although a following aerobic biological treatment step
was always necessary for compliance with effluent limitations.24 – 27
The idea of segregating domestic sewage to its black and gray
fractions inevitably involved new and compatible approaches for
treatment, utilization, and disposal of these fractions. Yet, surpris-
ingly enough, no information exists regarding the biodegradation
characteristics of the resulting sewage fractions. Consequently,
the prescribed treatment systems and techniques rely, not on
sound biodegradation characteristics as in the case of combined
domestic sewage, but on empirical approaches.28
In this context, the objective of this study was to investigate the
COD fractionation and biodegradation characteristics of black
and grey water components and compare them with those
determined for the combined domestic sewage. Respirometric
analyses based on the oxygen uptake rate (OUR) profiles obtained
for the two wastewater components and model calibration
of the experimental respirometric data was used for this
1242
purpose.
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c 2010 Society of Chemical Industry
S Murat Hocaoglu et al.
MATERIALS AND METHODS
Composition of segregated wastewaters
Raw black and grey water have been segregated and collected
from lodging houses and guesthouses of Tubitak MRC Campus.
Two buildings comprising a total of 28 apartments, 17 lodging
houses for permanent residents and 11 guest apartments for
temporary use were connected to separate holding tanks. Kitchen
wastewater stream was included in grey water. Coarse and
fine particles were removed from grey and black water with
a sequence of two 6 mm and 3 mm screens installed in the
corresponding holding tanks. Mixing was provided by pump
circulation. Wastewater samples were taken after the screening
unit.
The survey was conducted as the initial part of a comprehensive
study, which investigated the fate of non-biodegradable organics
and nitrogen removal dynamics in a membrane bioreactor (MBR)
system.29,30
Estimation of wastewater flow and load distributions
The daily flow rates for grey and black water were measured by
means of tipping buckets installed into the separate manholes. In
each setup, the numbers of turnovers were counted using a data
logger through a reed contact, yielding the flow rates of the two
wastewater fractions. The total wastewater flow rate was verified
with the total water consumption measured by water meters
connected to the buildings. The number of inhabitants staying
in the guesthouses was recorded daily. These records enabled
determining pollutant loads of black and grey water per person
and the amount of pollutants for segregated fractions.
COD fractionation
Soluble non-biodegradable COD fractions of grey and black water
were experimentally determined according to the procedure
proposed by Orhon et al.31 For each wastewater, two reactors were
run, one fed with raw wastewater and the other with wastewater
filtered using a 0.45 µm filter. The reactors were seeded with
small amounts of biomasses taken from the MBRs treating the
same wastewaters. To prevent nutrient limitation, nutrient and
buffer solutions were added prior to test. The reactors were
continuously aerated for about 45 days to enable the reduction
of all biodegradable organics and mineralization of the biomass.
Final threshold values of the total and soluble COD (Ct and St ) were
measured and evaluated as defined in the adopted procedure.
Biodegradable COD fractions were determined through respiro-
metric tests and mass balance. Respirometric tests were performed
with a Ra-Combo 1000 (Applitek Co., Nazareth, Belgium) contin-
uous respirometer. The biomass seeds for the reactor were taken
from the MBR treating black water. Respirometric tests were con-
ducted under similar conditions in the MBRs, with the same food to
microorganism ratio of 0.012 g COD g−1 VSS. The MLVSS concen-
tration in the black water MBR was about 5000 mg L−1 and sludge
age was 20 days. The respirometer consisted of a 3 L aerated reac-
tor and 750 mL chamber, in which dissolved oxygen consumption
in the mixed liquor was continuously monitored. The test was
started with the measurement of OUR rate of biomass to obtain
the initial endogenous respiration level. Afterwards, the wastew-
ater was added to the biomass in the reactor. The respirometric
OUR data were continuously recorded online. A dissolved oxygen
concentration higher than 3 mg L−1 was provided in the reactor
during the tests to avoid rate limitation. The bulk temperature was
kept at 20 ◦ C during the respirometric experiments. A nitrification
J Chem Technol Biotechnol 2010; 85: 1241–1249
COD fractionation and biodegradation of segregated domestic wastewater
Table 1. Experimental conditions of OUR tests
Initial Total Substrate to
wastewater reactor biomass ratio,
Wastewater volume, volume, So/Xo(mg COD
Type VWW (mL) VT (mL) (mg MLVSS)−1 )
Black water 100 2000 0.011
Grey water 500 2700 0.010
Combined water 200 2200 0.012
inhibitor (Formula 2533TM, Hach Company) was used to prevent
nitrification interference. It should be noted that grey water is by
nature nitrogen deficient, as also observed in the study. It is well
known that nitrogen deficiency may exert a limiting action on
biochemical reactions and therefore it runs the risk of masking
the true biodegradation characteristics of this sewage fraction, as
far as organic carbon is concerned. Consequently, ammonia was
added to secure excess nitrogen for the biochemical reactions.
On the other hand, nitrification inhibitor was added, as routinely
done in similar studies, to prevent nitrification interference in the
OUR profile, so that biodegradation characteristic of the organic
substrate (COD) fractions could be correctly ascertained. The com-
bined wastewater was prepared by mixing grey and black water
with the ratio of 1 : 1, which was determined according to the flow
rate of grey and black water on the day of experiment. The details
of the experimental conditions of OUR tests are given in Table 1.
The OUR data were analyzed by calibration of the selected
model, an Activated Sludge Model No.1 (ASM1) version modified
for endogenous decay.32 The same model was successfully used
in a number of similar characterization studies.33,34 The AQUASIM
simulation program (Swiss Federal Institute for Environmental
Science and Technology, Switzerland) was used in model
evaluations.35 The kinetic and stoichiometric parameters of the
model were estimated according to the procedure proposed by
Insel et al.13 using the SIMPLEX algorithm. The total biodegradable
COD (CS ) and its fractions, namely, the readily biodegradable
COD (SS ), the rapidly hydrolysable COD (XS1 ), and the slowly
hydrolysable COD (XS2 ) were calculated using the amount of
oxygen utilized.
The relevant chemical analyses for conventional characteriza-
tion were performed as defined in Standard Methods,36 with the
following codes for the corresponding procedures : SM- 5220 B
for COD, SM- 5210 B for BOD5 , SM- 2540 D for TSS, SM- 2540 E
for VSS, SM- 4500 for T.P., SM- 4500 B,C, for NH4 -N, SM- 4500-
B for TKN and SM-2320 B for alkalinity. The relative uncertain-
ties involved with the analytical procedures were assessed as
1.5–4% (50–500 mg L−1 , 500–1500 mg L−1 ) for COD, 15% for
BOD5 , 1–2% (0.5–10 mg L−1 , >100 mg L−1 ) for NH4 -N, 3.5–4.5%
for TKN (1–10 mg L−1 , >100 mg L−1 ), 4% for TP, and 2% for TSS
and VSS and 0.1 mg L−1 h−1 for the OUR measurements.
RESULTS AND DISCUSSION
Composition of segregated wastewaters
Conventional characterization of segregated wastewater streams
was performed on daily composite samples collected around twice
a week for about 18 months, to have full coverage of daily and
seasonal variations. Characteristics in terms of major parameters
with average concentrations and their standard deviation are
given in Table 2, together with relevant characterization data
J Chem Technol Biotechnol 2010; 85: 1241–1249

c 2010 Society of Chemical Industry
www.soci.org
from the literature. The main observations concerning the organic
pollutants, derived from the presented data may be summarized
as follows. (i) Segregation of wastewater streams generated, as
typically reported in the literature, a strong black water fraction
and a much weaker grey water fraction with average COD values
of 1225 mg L−1 and 295 mg L−1 , respectively. (ii) The mixture
represented a relatively strong domestic sewage with a COD value
of around 650–700 mg L−1 , showing that sewage characteristics
are quite site-specific. In fact, in a number of studies carried out
in different parts of the Istanbul Metropolitan Area, the average
COD level of the sewage was observed to vary in the wide range
315–680 mg L−1 .8,31,37 – 39 A closer COD value of 679 mg L−1 was
reported for the municipal sewage of a small town adjacent to
the study area40 (iii) Segregation imparted markedly different size
distributions for organic pollutants: while the COD content of
black water was predominantly particulate (67%) as differentiated
with the routine filtration size of 450 nm, COD in grey water was
mainly (65%) soluble. Hence, the hydrolysis process appears to be
a more significant step for the biodegradation of black water. This
observation was also supported by corresponding values of total
suspended solids (TSS) and volatile suspended solids (VSS) in the
two wastewater streams. (vi) BOD5 /COD ratios were computed as
0.28 for black water and 0.38 for grey water, lower than the ratio of
0.44 reported for Istanbul wastewaters.38 This may be interpreted
as a good indication for the unreliable nature of BOD5 in reflecting
the biodegradation trend of the organic matter in wastewaters.
Table 2 also indicated that the majority of nitrogen and
phosphorus were collected in black water. The average COD/N/P
ratio, commonly used as a rough preliminary index for nutritional
requirements in biodegradation, was calculated as 100/2.5/2.5
for grey water. This ratio, although supported by the findings of
Almeida et al,41 is lower in nitrogen and higher in phosphorus than
other similar data in the literature;42 it mainly indicates nitrogen
limitation for effective performance of biological treatment. The
high phosphorus ratio can be explained by phosphorus-containing
soaps and detergents in grey water, possibly attributed to the daily
cleaning and laundry of the guesthouses.
The same COD/N/P ratio can be calculated as 100/14.7/2.0
for black water, corresponding to an average COD/N ratio of
about 6.8, a level unsuitable for efficient denitrification without
external carbon source.43 Furthermore, when high TKN values
are considered with relatively lower alkalinity concentrations in
black water, nitrification limitation may also be a problem. In
this context, black water poses an interesting problem for an
appropriate engineering approach, with its high nitrogen content
on the one hand and low organic carbon and alkalinity contents
on the other hand. Biological nitrogen removal, if needed, would
require systems capable of offering simultaneous nitrification
and denitrification (SNdN) and a well controlled and balanced
nitrification/denitrification scheme.44 – 46
Estimation of wastewater flow and load distributions
The average number of people staying in the buildings was 21;
it varied from 18 to 26 during the evaluation period. Grey and
black water flow rates coming to the pilot plant and total water
consumption were measured for a month. Grey water flow rate was
between 1200 and 2350 L day−1 , whereas it was 1350 to 3400 L
day−1 for black water. The daily total wastewater generation
was calculated as 197 L per person. Overall, about 60% of the
total household wastewater was black water. This value coincides
with the study done by Cossu et al.46 However, it is in contrast
1243
to most studies as they reported a higher ratio of grey water
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 www.soci.org S Murat Hocaoglu et al.

Table 2. Characteristics of black and grey water

This study

Black water Grey water

Black water42
Mean/ Mean/ (low Grey water49 Grey water50 Domestic Domestic
(Number of Std. (Number of Std. flushing wc) Black mainly shower - waste- waste-
Parameters samples) Dev. samples) Dev. range water48 bathroom Morocco water40 water38

PH 8.0 (105) 0.3 7.2 (103) 0.3 8.87-9.08 8.02 6.4-8.1 7.4 – 7.2
Alkalinity CaCO3 , mg L−1 679 (60) 87 192 (23) 29 – – 24-43 – – 300
BOD5 , mg L−1 338 (55) 155 111 (55) 33 410-1400∗ 458 76-200 90 – 180
Total COD, mg L−1 1225 (109) 560 295 (105) 79 806-3138 1013 – 180 679 410
Soluble COD, mg L−1 407 (67) 119 191 (64) 54 – – – – – 140
TKN, mg L−1 180 (98) 28 7.4 (96) 3.7 130-180∗∗ 41 4.6-20 – 97 43
NH4 + -N, mg L−1 147 (89) 18 1.6 (86) 1.4 – 21 <0.1-15 12 77 30
TP, mg L−1 25 (76) 9 7.3 (77) 3.1 21-58 11 0.11-1.8 2.0 – 7.2
TSS, mg L−1 625 (102) 437 63 (103) 30 920-4320 715 48-120 200 334 210
VSS, mg L−1 529 (100) 377 47 (100) 23 420-3660 401 – – 261 195
∗
BOD7 , ∗∗ T.N.

Table 3. Daily nutrient and organic matter loads of black and grey water per capita

Black water53
This study Black (from food supply data)
water51 Light grey Black
Parameter Black Grey Combined Domestic typical water-grey Black water26 Black
g p−1 d−1 water water wastewater wastewater51 load water51 water52 Sweden China South Africa India Europe water54

TN 19.6∗ 0.7 20.3 2-15 12.05 1.1-1.92 11.93 12.60 10.95 9.31 20.27 8.5-13 12.5
NH4 + -N 17.8 0.2 18 – – – – – – – – – –
TP 3.7 0.8 4.5 1-3 1.92 0.27-0.55 1.82 1.37 1.64 1.37 3.01 0.9-1.7 1.4
COD 90.0 24.9 114.9 25-200 73.97 10.1-54.8 48.49 – – – – 55.7-66.5 –
BOD5 31.1 8.7 39.7 15-80 24.66 4.93-35.6 – – – – – 19-39.5 –
Potassium – – – 3.56 – 3.25 3.84 1.8 4.38 4.11 – –
TSS 37 6.0 43 – – – – – – – – – –
VSS 33 4.5 37.5 – – – – – – – – – –
∗
TKN.

production. During the evaluation period, grey water was close to 100%
between grey and blackwater,

the typical grey water characteristics, on the other hand, slightly

Distrubution of pollutants

80%
diluted characteristics were observed for the black water, which
was consistent with the high water production ratio of black 60%
water. Water consumption per toilet flushing was measured as
40%
around 9 L. It should be noted that the apartments were originally
constructed with conventional plumbing. A separate new pipeline 20%
was constructed and wastes of all of toilets were connected to this
line for black water collection. The existing plumbing was used for 0%
BOD COD TKN NH4-N T.P SS VSS
grey water collection. Parameters
Daily nutrient and organic matter loads of grey water and black Blackwater Greywater
water per capita per day and comparison with the literature are
given in Table 3. Overall, the daily loads of black water per capita Figure 1. Load distributions of segregated household wastewater frac-
tions.
are slightly higher than the values in the literature. However,
the ratio of organic matter and nutrients are in the range of the
values given by Kujawa-Roeleved and Zeeman26 for Europe, and The load distributions between grey and black water in
by Henze and Ledin.51 It is reported that wastewater composition household wastewater are illustrated in Fig. 1. More than 80%
depends on the lifestyle and habits of the population and the of organic matter, suspended solids, total phosphorous and most
nutrient content of the resulting wastewater varies among men, of the nitrogen (99% of the NH4 -N) in household wastewater
1244

women, and children in the same country.17,26 comes from toilets.

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c 2010 Society of Chemical Industry J Chem Technol Biotechnol 2010; 85: 1241–1249
COD fractionation and biodegradation of segregated domestic wastewater www.soci.org

(a) (b) 3.5

3.5
3.0 3.0

2.5 2.5

L/person
L/person
2.0 2.0
1.5 1.5
1.0 1.0
0.5 0.5
0.0 0.0
00:08
01:53
03:38
05:23
07:08
08:53
10:38
12:23
14:08
15:53
17:38
19:23
21:08
22:53

00:08
01:53
03:38
05:23
07:08
08:53
10:38
12:23
14:08
15:53
17:38
19:23
21:08
22:53
Time (h) Time (h)

Figure 2. Grey and black water generation per person (average for 1 month) (a) grey water (b) black water.

The grey and black water generation per person for 24 h cycle
Table 4. Experimental assessment of soluble residual COD fractions
are illustrated in Fig. 2. The data shown in the figure were obtained
by averaging the flow rate collected every 15 min over 1 month. Black Grey Combined
For both fractions, wastewater generation increased during the COD fractions water water wastewater
early in the morning (06 : 30–10 : 30) and throughout the evening Initial total COD, CT (mg L−1 ) 1145 275 665
period (18 : 00–23 : 00). Initial soluble COD, ST (mg L−1 ) 360 210 280
Initial soluble inert COD, SI 42 14 30
COD fractionation (mg L−1 )
Respirometric measurements were conducted separately to obtain SI /CT (%) 3.7 5.0 4.5
the OUR profiles of black water and grey water, as well as the Initial biodegradable COD, CS 1088 259 627
(mg L−1 )∗
combined wastewater reflecting the flow proportionate mixture
YSP = SP /CS 3.5 3.4 3.5
of the two streams. The OUR represents the process rate of
Soluble inert microbial 38 9 22
oxygen utilization and dissolved oxygen, SO , is a significant model products, SP (mg L−1 )
component in current models.6,13 The OUR profiles generated
∗ From respirometric measurements.
during the respirometric experiments were used to yield the
magnitude of the total biodegradable COD, CS for the tested
wastewater samples. In fact, the OUR area above the endogenous
respiration level gives the amount of oxygen,
O2 , corresponding by calibration of the selected model with the experimental OUR
to available biodegradable COD.11,55 profiles associated with the wastewaters. The calibration identified
aside from the readily biodegradable fraction, SS , two hydrolysable

O2 = CS (1 − YH ) COD fractions, XS1 and XS2 for black water and only a single – slowly
biodegradable COD fraction, XS2 – for grey water. The results
where, YH is the heterotrophic yield coefficient. In the respirometric obtained are outlined in Table 5, together with two similar data
experiments, CS values were computed for a selected YH of 0.67 g for local municipal sewage.8,40
cell COD g−1 COD, which was validated with model calibration of Two significant observations should be underlined, concerning
the OUR data. the results displayed in Table 5. (i) The relative magnitude of the
The difference between the total COD (CT ) and the biodegrad- readily biodegradable COD fraction, SS was high compared with
able COD (CS ) gives the total residual COD (CI ) covering the soluble typical domestic sewage, especially for grey water, where model
and particulate residual COD fractions, SI and XI , respectively. As calibration yielded an unusually high SS /CT ratio of around 30%;
previously mentioned a direct procedure was used for the assess- this may be due to the restricted household characteristics of
ment of the soluble residual COD yielding aside from SI , soluble the selected residential community. The corresponding SS fraction
inert microbial products, SP in terms of a yield coefficient YSP , may also include a portion with a high hydrolysis rate – very close
which expresses SP as a fraction of the total biodegradable COD to substrate utilization rate for microbial growth – not separately
in the wastewater.37 The results of three different experiments identifiable through the resulting OUR profile. (ii) The level of
conducted on samples approximating the average characteristics particulate inert COD fraction, XI , was relatively low, because the
of black and grey waters were converted to the respirometric slowly hydrolysable fraction is likely to include some of XI based
experiments as shown in Table 4. on the hydrolysis rates obtained, which will be explained in more
The particulate inert COD fraction of wastewater, XI was detail in the following section.
computed from mass balance:
Biodegradation characteristics
X I = CT − CS − S I Model calibration yielded a close simulation of the experimental
OUR profiles as illustrated in Figs 3 and 4. Kinetic coefficients
The repartition of the biodegradable COD to total COD derived from model calibration are listed in Table 6 together
1245

fractions with different biodegradation characteristics was done with selected relevant data from the literature; they provided

J Chem Technol Biotechnol 2010; 85: 1241–1249

c 2010 Society of Chemical Industry www.interscience.wiley.com/jctb
 www.soci.org S Murat Hocaoglu et al.

Table 5. Results of COD fractionation for tested wastewaters and local municipal sewage
Black Grey Combined Domestic Domestic
COD fractions water water wastewater wastewater8 wastewater40

Total COD, CT , mg L−1 1145 275 665 450 679

Soluble COD, ST , mg L−1 360 210 280 132 224
ST /CT % 31.6 76.4 42.0 29.3 32.9
Particulate COD, XT , mg L−1 782 66 386 318 401
XT /CT % 68.4 23.6 58 70.6 59.1
Biodegradable COD, CS , mg L−1 1088 259 627 348 584
CS /CT , % 95.1 93.5 94.1 77.3 86.0
Readily biodegradable COD, SS , mg L−1 168 81 121 100 251∗
SS /CT , % 14.7 29.3 18.2 22.2 36.9
Rapidly hydrolysable COD, XS1 , mg L−1 340 – 121 116 –
XS1 /CT , % 29.7 – 18.2 25.7 –
Slowly hydrolysable COD, XS2 , mg L−1 580 178 385 132 333
XS2 /CT , % 50.7 64.2 57.8 29.3 49.0
Soluble inert COD, SI , mg L−1 43 14 30 32 27
SI /CT , % 3.7 5.0 4.5 7.1 3.9
Particulate inert COD, XI , mg L−1 13 4 9 70 68
XI /CT , % 1.1 1.5 1.3 15.6 10.0
∗ Rapidly hydrolysable COD included.

(a) (b)

Figure 3. Biodegradation characteristics of black water: (a) model calibration of the OUR profile; (b) simulated biodegradation profiles of COD components.

(a) (b)

Figure 4. Model calibration of the OUR profiles for (a) grey water (b) combined wastewater.
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COD fractionation and biodegradation of segregated domestic wastewater www.soci.org

Table 6. Assessment of model parameters by means of OUR calibration

Black Grey Combined Filtered Settled Domestic
Model parameters & state variables Unit water water wastewater sewage58 sludge58 sewage57

Maximum growth rate for XH , µHmax day−1 4 3.8 4 4 4.2 3.5

Half saturation constant for growth of XH , KS mg COD L−1 4 7 4 3 3 2
Maximum hydrolysis rate for XS1 , kh day−1 2.8 – 2.8 1.7 1.7 2
Hydrolysis half saturation constant for XS1 , KX g COD (g cellCOD)−1 0.03 – 0.03 0.05 0.05 0.2
Maximum hydrolysis rate for XS2 , khx day−1 1.1 1.1 1.1 – 0.70 –
Hydrolysis half saturation constant for XS2 , KXX g COD (g cellCOD)−1 0.1 0.03 0.1 – 0.05 –
Endogenous decay rate for XH , bH day−1 0.18 0.18 0.18 0.20 0.20 0.24
Heterotrophic yield coefficient, YH g cellCOD (g COD)−1 0.67 0.67 0.67 0.67 0.67 0.67
Total active biomass mg COD L−1 3710 (%52) 3555 (%50) 3470 (%49) – – –

a specific fingerprint for the biodegradation characteristics of
studied wastewater streams. Major features of the results obtained
may be underlined as follows. (i) The biodegradation of the readily
biodegradable COD, SS was characterized with a maximum specific
heterotrophic growth rate, µH , of around 4.0 day−1 a typical value
within the range reported for domestic sewage. (ii) As previously
mentioned, model calibration identified two COD fractions with
different hydrolysis rates for black water. This confirms similar
observations in the literature arguing that the hydrolysable COD
portion constitutes the bulk of the COD content in domestic
sewage and is therefore, too large to be accurately defined with
a single hydrolysis mechanism. Orhon et al.56 reported a rapid
hydrolysis rate of 3.1 day−1 and a slow hydrolysis rate of 1.2 day−1 ,
supporting the results of this study. (iii) Black water includes the
settleable portion of the COD. Okutman et al.57 suggested that
settleable COD be accounted for as a separate COD fraction
because it was associated with a slower hydrolysis mechanism
than the rest of the hydrolysable COD. Orhon et al.58 reported
a hydrolysis rate of 0.7 day−1 for the settleable COD as given in
Table 6. In this context, the khx value of 1.1 day−1 obtained in
this study may be interpreted as an average level representing
the hydrolysis of the slowly hydrolysable and settleable COD
fractions in black water. This rate is quite slow and extends
beyond the experimental OUR analysis as shown in Fig. 4(b) giving
indications that the corresponding XS2 fraction may well cover
a part of the non-biodegradable particulate COD, XI . This may
be an acceptable explanation for the low level of XI observed
in the study. (iv) Grey water was only associated with the slowly
hydrolysable COD fraction, XS2 with the same slow hydrolysis rate
as the corresponding part in black water. This observation and the
high level of its readily biodegradable COD content, as previously
given in Table 5, suggest that a part of Ss may also include rapidly
hydrolysable COD with a high hydrolysis rate compatible with
the utilization of SS . In fact, each COD fraction inevitably includes
a wide range of different organic compounds with compatible,
yet possibly slightly different characteristics and only average
biodegradation characteristics can define this fraction. This is
reflected in the model adopted, which gives a simplified but
acceptable account of these characteristics.
When domestic sewage sources are segregated, organic
substances in the grey water fraction come from kitchen
wastewater (food residuals, spoiled juices, spoiled foods/drinks
etc.), from bath, shower (soaps, shampoos, body dirt, dead skin,
etc.), from laundry (detergents, softeners and dirt on cloths, etc.).
Therefore, the nature and particle size distribution of pollutants
It is then quite conceivable that a small portion within the readily
biodegradable and readily hydrolysable COD fractions overlap
in terms of biodegradation characteristics. In other words, the
rate of direct utilization for microbial growth and that of rapid
hydrolysis approach one another and become compatible for this
COD portion. In this case, identification and tagging of this COD
portion in one or the other fraction does not involve an evaluation
error because the emphasis remains on utilization kinetics for both
COD fractionation and the corresponding modeling.
Analysis of the biodegradation characteristics of the combined
sewage, which was segregated into its black and grey water
components, was also included in the study. Related results given
in Figs 3 and 4 and Tables 5 and 6 provide a useful basis for the
possible impact of stream segregation on resulting biodegradation
characteristics. First, the results confirmed the site-specific nature
of the evaluation as the characteristics of the sewage tested
in the study showed a number of slight differences with those
reported in the literature. The significant feature of the evaluation
was the observation that stream segregation did not impart
an appreciable variation in the biodegradation characteristics,
when interpreted in terms of COD fractionation and kinetics of
major biochemical processes. The COD fractions displayed in
Table 5 indicated a similar profile for soluble COD components
and significant accumulation of the particulate COD components
in the black water fraction, which also incorporated the majority of
nitrogen and phosphorus. Utilization of the readily biodegradable
COD for microbial growth and dual hydrolysis took place, as
shown in Table 6, with the same rate coefficients regardless
of the wastewater stream so that the biodegradation pattern
solely depended on the repartition of the COD fractions between
black and grey water streams. The results indicated that aerobic
treatment may prove equally effective for the two wastewater
streams.
CONCLUSION
COD fractionation and assessment of process kinetics by model
calibration of respirometric data provided a rational and scientific
basis for evaluating biodegradation characteristics of black and
grey water streams.
For the wastewater tested in the study, stream segregation did
not affect biodegradation characteristics of black and grey water
fractions. Process rate coefficients for the utilization of the readily
biodegradable COD and for the hydrolysis of the rapidly and slowly
hydrolysable COD fractions remained the same after segregation
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significantly change and accumulate within the soluble range.30 and very close to those reported for typical domestic sewage.

J Chem Technol Biotechnol 2010; 85: 1241–1249

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 www.soci.org
Therefore, the difference between the removal rates of
biodegradable COD in black and grey waters originated not from
process kinetics, but from the repartition of different COD fractions,
accumulating the more slowly biodegradable components in black
water and increasing readily biodegradable COD components in
grey water.
The results suggest aerobic treatment as an equally effective
biological treatment scheme for both wastewater streams. A
high rate aerobic process efficiently removing the soluble COD
components will potentially generate a nutrient-rich effluent for
reuse and energy-rich sludge for recovery.
ACKNOWLEDGEMENT
The authors gratefully acknowledge support provided through the
Zer0-M project financed by the European MEDA Water program
(ME8/AIDCO/2001/0515/59 768) for the experimental set up used
in this study.
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