LABORATORY REPORT

LAB 8: Chemical Oxygen Demand

LAB 9: Disolved Oxygen & Biological Oxygen Demand

SUBMITTED BY:
MR. BISHAL BHARI (115461)
Group Members
Ms. Suchanya Wongrod (115474)
Ms. Nguyen Nhat Huynh Mai(115488)

SUBMITTED TO:
DR. OLEG SHIPIN
MISS SALAYA PHUNSIRI

ED 78.01: ENVIRONMENTAL CHEMISTRY AND LABORATORY

ENVIRONMENTAL ENGINEERING AND MANAGEMENT
SCHOOL OF ENVIRONMENT, RESOURCES AND DEVELOPMENT
ASIAN INSTITUTE OF TECHNOLOGY

Chemical Oygen Demand October 30, 2013

1. Introduction
The chemical oxygen demand (COD) is a measure of the oxygen equivalent of the organic
matter in a sample that is susceptible to oxidation by a strong oxidizing agent. This test is widely
used to measure the organic strength of domestic and industrial wastewater, and it can be related
empirically to BOD, organic matter and organic carbon. In short, COD measure those substances
which can be chemically oxidized. The main advantage of the COD method is that it is rapid; it
takes only a few hours to complete compared to 5 days for BOD. The COD of wastewater is
generally higher than the BOD because more substances can be chemically oxidized than can be
biologically oxidized. It is possible to correlate COD with BOD for many types of wastes, and
the COD test can then be used as a quick substitute for BOD. Also, wastewaters containing
compounds toxic to microorganisms can be analyzed for COD.
Furthermore, the COD method yields more reproducible results than the BOD method. One
disadvantage of the COD test is that it cannot differentiate between biologically oxidisable and
biologically inert organic matter. It is therefore less relevant to natural processes than the BOD
test. Additionally, COD values can characterize the degree of pollution and self-purification of
various waters. National and international standards and guidelines have been set for COD. The
COD test is useful for monitoring and control after correlation with sources of organic matter has
been established.
Generally, COD values are higher than BOD values. The reason is that biochemical oxygen
demand measures only the quantity of organic material capable of being oxidized, while the
chemical oxygen demand represents a more complete oxidation. Typical COD values for
domestic waste range from 200 - 500 mg/l.

2. Objective
a) To determine the Chemical Oxygen Demand (COD) value of the sample by Closed
reflux method.
b) To prediction the relationship between COD values and the probable BOD5 of sample.

3. Methodology
3.1 Principle
The given sample was refluxed on strongly acid solution with a known amount of K2Cr2O7. After
digestion the remaining unreduced K2Cr2O7 was titrated with FAS to determine the amount of
K2Cr2O7 consumed. Oxidizable organic matter was then calculated in term of O2 equivalent

Chemical Oygen Demand October 30, 2013

3.2 Procedure
The sample was diluted to 1:50 and 1:100

10 mL of diluted sample was then placed in the culture tube (25 x 150 mm)

6 mL of standard digestion solution (K2CrO7) was then added in the tube

14 mL of H2SO4 + Ag2SO4 reagent was carefully ran down inside of vessel

The cap tubes were tighten and the solution was mixed completely

The tube was then kept in 150oC oven and refluxed for 2h

It was then cooled to room temperature

1-2 drops of ferroin indicator was added in it

It was then titrating with 0.10 M FAS (Color change from blue-green to reddish brown)

Blank Determination was also then done

COD was then calculated

3.3 Calculation
COD, mgO2/L = (A-B) x M x 8000
mL of sample
Where,

4.

mL of FAS used for blank

mL of AFS used for sample

Molarity of FAS

Presentation of Results

4.1 COD calculation

Table 1: COD titration of sample no-CDB10
Sample No: CDB10
Blank
Sample

Dilution
1:50
Dilution

Titration
No
1
2
1
2
1

Volume
of FAS
6.0
5.9
4.5
5.85
5

Average

Remark

5.95
This dilution result was
rejected due to bias result
5.125

Chemical Oygen Demand October 30, 2013

1:100

5.25

Calculation
Calculation:
COD, mg O2/L

(A-B) x M x 8000 * DF
ml of sample

Where:
A = ml of FAS used for blank
B = ml of FAS used for sample
M = Molarity of FAS
DF= Dilution Factor

COD, mg O2/L

(A-B) x M x 8000 * DF
ml of sample

(5.95 - 5.125) x 0.1 x 8000 * 100

10 ml
6600 mg O2/l

=
4.2

Probable BOD
Given :
BOD/COD = 0.4
Therefore,
Estimated BOD = 6600 x 0.4 = 2640 mg/l

5. Discussion of Result
COD value of the sample CBD10 was calculated to be 6600 mg O 2/L which is quiet high
concentration. This value is defined as the amount of a specified oxidant that reacts with the
sample under controlled conditions. The quantity of oxidant consumed is expressed in terms of
its oxygen equivalence.
This result is used to estimate BOD value of the unknown sample. COD measurement helps to
determine the fraction of dilution of BOD. The typical ratio of COD to BOD for this sample is
0.4 hence the probable BOD can be calcualted to be 2640 mg/l.
3

Chemical Oygen Demand October 30, 2013

COD and BOD value is important to decide the wastewater treatment process. If the ratio of
BOD to COD is much higher than 0.5 so the biological treatment process should be used.
Similarly if the amount of COD is higher than amount of BOD such as some specific industries
that use for wastewater contain high amount of chemical, the chemical treatment process should
be selected.
The results from analysis might have some error due to following factors:
i.

If there are some organic substances, such as molecule fatty acid which can not be
oxidized by Cr2 O7-2, this will result in higher COD value than the exact value.

ii.

If the sample consists of chloride and nitrite, it can react with the Cr 2O72-. This can cause
the COD value to be higher than exact value. However HgSO 4 and sulfamic acid was
used to get rid of them.

iii.

FAS concentration is not stable

iv.

Error due to cleaning operation: Improperly cleaned tubes and cap can also result in
erroneous result.

v.

Oven temperature is required to be 150 degree celcius for 2 hour. However, the oven
operation by the entire group was not syncronised and it was opened and closed several
time during the 2 hour interval. This might result in erroneous result.

References

AIT, SERD, EEM, Course ED 78.01-Environmental Chemistry and Laboratory,

Laboratory Instruction Sheet 8: Chemical Oxygen Demand;2013

Sawyer C.N et.al, Chemistry for Environmental Engineering: 5th edition Mc.GrawHill;2002

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

1. Introduction
Dissolved oxygen is the measure of the amount of gaseous oxygen (O 2) dissolved in an aqueous
solution. Dissolved Oxygen determinations are usually used for checking water quality of rivers
and wastewater from wastewater treatment plant, checking the dissolve oxygen of organic matter
in the plant. Moreover, Dissolve Oxygen determination is a basic of BOD test.
D.O. can be measured by Winkler titration. Measurements of D.O. can be made more
conveniently with electrochemical instrumentation. "D.O. meters" are subject to less interference
than the Winkler titration. They are portable and can be calibrated directly by using the oxygen
in the air.
DO is operational parameter in wastewater treatment process. Sufficient D.O. is essential for the
proper operation of many wastewater treatment processes. Activated sludge tanks often have
their D.O. monitored continuously. Low D.O values may be set to trigger an alarm or activate a
control loop which will increase the supply of air to the tank.
Biochemical Oxygen Demand (BOD) is defined as the amount of DO consumed in 5 days when
the waste is incubated in the dark at 20 0C in the presence if suitable stabilization microorganism.
It is the oxygen required by bacteria for stabilizing decomposable organic matter under aerobic
conditions. The BOD test is a bioassay procedure involving the measurement of oxygen
consumed by living organisms while utilizing the organic matter present in a waste, under
conditions as similar as possible to these that occur in nature. The BOD test may be considered
as a wet oxidation procedure in which the living organisms serve as the medium for oxidation of
the organic matter to carbon dioxide and water. The oxidation reaction involved in the BOD test
is a result of biological activity, and the rate at which the reaction proceeds is governed to a
major extent by population numbers and temperature. Temperature effects are held as far as
natural bodies of water are concerned. Normally, we use BOD5 or the 5 days incubation period
because we need to minimize interference from oxidation of ammonia. During 5 days, BOD
values follow first order reaction. The BOD test is used to evaluate the pollution strength of
domestic and industrial waste in the terms of Oxygen that they will require if discharge into
natural watercourses in which aerobic condition exist.

2. Objective
i.

To determine DO and BOD of sample by Azide Modification Method.

ii.

To understand the realtionship of DO, BOD and COD

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

3. Methodology
DO Determination (Azide Modification Method)

O.2, 0.5 and 1 mL of sample was added in each BOD bottles of size 300 mL using the
following table
Table: BOD measurable with various dilution of sample
Using percent mixture

By direct pipette into 300-ml bottles

% Mixture

Range of BOD

ml

Range of BOD

0.01
0.02
0.05
0.10
0.20
0.50
1.00
2.00
5.00
10.00
20.00
50.00
100

20,000 70,000
10,000 35,000
4,000 14,000
2,000 7,000
1,000 3,500
400 1,400
200 700
100 350
40 140
20 70
10 35
4 14
0-7

0.02
0.05
0.10
0.20
0.50
1.00
2.00
5.00
10.00
20.00
50.00
100
300

30,000 105,000
12,000 42,000
6,000 21,000
3,000 10,500
1,200 4,200
600 2,100
300 1,050
120 420
60 210
30 105
12 42
6 21
0-7

Supersaturated oxygen distilled water was shiponed up to 300 mL and its cap was closed

BOD sample was then incubated for 5 days (DO5) in refrigerator at 20oC

Samples for determine DO0, add 1 mL MnSO4 of Alkali Iodide-Azide. Flocculation

occur. Mix by inverting a few times.

Let the manganese hydroxide flocs settle to half of the bottle.

Add 1 ml conc. H2SO4 (allowing the acid to run down the neck of the bottle) and mix
several times till precipitate disappear.

99 ml of the solution was removed

The remaining solution of 201 ml was titrated with 0.025 M Na2S2O3 to pale straw color.

Few drops of starch solution was added and it was immediately titrated from blue color to
colorless.

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

Blank determination was done in the same manner by using only supersaturated oxygen
distilled water.

Calculation BOD
BOD5 (mg/l) =

(D1 - D2) - (BOD of Blank sample)

P
Where,D1 = DO of diluted sample after preparation, mg/L
D2 =DO after 5 days incubation at 20oC, mg/l.
P =Decimal volumetric fraction of sample used.

4.

Presentation of Results

4.1

BOD5 of Sample CBD10

Volume of Na2S2O3

Dilution

Volume
of sample
(ml)

BOD5

7.6

0.2/300

1200

7.6

0.2/300

1200

7.2

0.5/300

720

6.8

0.5/300

960

6.55

1/300

555

8.2

6.2

1/300

660

8.4
8.4

8.8*
8.4

300/300
300/300

DO0
(mg/l)

Avg
10

0.2

20

0.5

Blank

8.4
8.8
*
8.5
9.8
*
8.2

Remark

DO5
(mg/l)

8.4

Rejected as
depleted
DO is less
than 2 mg/l
Can be
used

*Over titration hence the data cant be used

Calculation:
BOD5 (mg/l) =

(D1 - D2) - (BOD of Blank sample)

P
Where,D1 = DO of diluted sample after preparation, mg/L
D2 =DO after 5 days incubation at 20oC, mg/l.
P =Decimal volumetric fraction of sample used.
For 1 ml. of sample: BOD5 =(8.4-6.2)
1/300
3

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

= 660 mg/l

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

4.2

BOD5 of all group

COD

BOD
COD

Calculated
BOD

Dilution

10

20

30

D
O0

DO5

10

20

BOD5

30

Remark

10

20

30

GR 1

7400

0.4

3000

0.2

0.5

8.4

5.3

6.2

4650

1320

720

All can be used

GR 2

8200

0.4

2880

0.2

0.5

8.3

7.8

7.3

5.8

600

600

750

10, 20 cant be used

GR 3

5680

0.4

2500

0.2

0.5

8.3

8.0

7.25

6.7

450

630

480

None can be used

GR 4*

6600

0.4

2640

0.2

0.5

8.4

7.6

7.2

6.2

1200

840

660

10, 20 cant be used

GR 5

6000

0.6

3600

0.2

0.5

8.5

7.7

7.10

6.25

1200

840

675

10, 20 cant be used

GR 6

5400

0.6

3240

0.2

0.5

8.7

8.6

7.55

6.5

75

720

660

10, 20 cant be used

GR 7

4000

0.6

2400

0.2

0.5

8.5

8.0

7.6

6.4

750

540

630

10, 20 cant be used

GR 8

4932

0.6

3000

0.2

0.5

8.6

8.5

7.55

6.55

675

630

615

10, 20 cant be used

GR 9

4053

0.8

3050

0.2

0.5

9.2

8.0

7.62

6.6

1500

1068

960

10, 20 cant be used

GR10

2880

0.8

2300

0.2

0.5

8.3

8.05

7.1

6.3

375

810

600

10, 20 cant be used

GR11

3600

0.8

2900

0.2

0.5

9.0

8.05

7.6

6.25

1425

820

825

10, 20 cant be used

* GR 4- CBD10 sample data

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

5. Discussion of Result
The BOD test was carried out after COD test. COD value was 6600 mg/L. Probable BOD of the
sample was calculated to be 2640 mg/l with the help of provided BOD to COD ratio of 0.4
established for the sample provided. Dilutions were then chosen as reference to lab instruction
sheet which were 0.5 mL, and also one above (0.2 mL) and one below (1 mL).
After 5 days of incubation, the BOD5 of the sample CBD10 was calculated to be 660 mg/l. Only
one result out of all the sample dilution could be used as only its DO delpletion was more than 2
mg/l. Hence the BOD5 was calculated to be much lower than the probable BOD which signifies
that the BOD to COD ratio was much lower than the 0.4 and the actual BOD to COD ratio of the
sample CBD10 can be calculated to be 0.1. Such a low ratio signifies the presence of inorganic
or resistent material thus we can conclude that the sample source cannot be biodegraded. Such a
low BOD to COD ratio also suggests the toxic nature of the sample.
Considering all the group data almost all the group had to use the 1 ml dilution DO 5 data for the
calulation of BOD5 hence the BOD5 of the entire groups lies between 600 to 2100 mg/l. The
BOD to COD ratio provided to the entire groups was much higher than the actual value. Hence
that was the reason that there lied a marginal gap between probable BOD and the actual BOD.
During the BOD test, errors can occur due to either human, instruments or the unpredictable
biological activities. Therefore, cares should be taken as follows:

Clean BOD bottle properly with detergent to eliminate all previous and accumulated
organic matter and microorganism. Rinse with distilled water and let dry to minimize
toxic from chlorine residue from tap water.

Some inhibitors, such as heavy metal and some toxic, can inhibit micro organism growth
or may kill micro organism.

Controlling of pH, oxygen content, and temperature of dilution water should be

maintained to standard. Dilution water should have pH in range of 6.5 8.3 and contain
necessary nutrient.

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

References

AIT, SERD, EEM, Course ED 78.01-Environmental Chemistry and Laboratory,

Dissolved Oxygen and Biochemical Oxygen Demand;2013

Sawyer C.N et.al, Chemistry for Environmental Engineering: 5th edition Mc.GrawHill;2002

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

Appendix
a)

Write chemical equations for all the essential reactions of the Winkler method.

1. In absence of oxygen, the added MnSO4 will form Mn (OH)2

Mn2+ + 2OH-

Mn(OH)2 (white solid)

2. In presence of oxygen, the added MnSO4 will form MnO2

2Mn (OH)2 + O2

2MnO2 (brown solid) + 2H2O

3. After adding conc. H2SO4

MnO2 + KI + 2H+ + SO42-

MnSO4

+ I2 + 2H2O + K2SO4

4. When titrating with Na2S2O3, Iodine will react with Na2S2O3 and form NaI.
2Na2S2O32- + I2

Na2S4O62- + 2 Na I

b) Explain why oxygen should be fixed in the field, when making determinations of DO in
streams, lakes, etc.
Water sample have bacteria that can degrade organic matter by using oxygen in water. This can
cause the reducing of dissolved oxygen. On the other hand, Oxygen in the air can penetrate into
water sample. Therefore, the best way to keep oxygen at the same amount of initial value is
fixing by MnSO4.
c) Why do you use Azide modification of the Winkler method?
Nitrite is one of the main interference involving DO measurement by Winkler method. During
titration process, nitrite oxidizes I- to I2, thus giving a very high result of DO. Nitrite can exist
in effluent from wastewater treatment plant, natural stream, lake, etc. The chemical reactions
involved in this process are as follows:
NaI + NO2-

N2O2 + I2

Further, N2O2 is oxidized by oxygen can give NO2 again. This can be a cyclic reaction.
N2O2 + 1/2 O2 + H2O

2 NO2- + 2 H+

To solve this problem, the modification method use sodium azide (NaN 3) to capture NO2, and
make it into N2 and N2O.

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

2NaN3 + 2H+ + SO42HN3 + NO2- + H+

2HN3 + Na2SO4
N2 + N2O + H2O

d) Draw up a table showing five substances which interfere with the Winkler test and
indicate the modification designed to overcome these interferences.
Interference
substances

Chemical
Used

Method

Chemical Reactions

NO2-

NaN3

SAM

NaN3 + H+ HN3 + Na+

HN3 + NO2- + H+ N2 + N2O + H20

kMnO4
RSM

5 NO2- + 2MnO4- + 6H+

5NO3- + 2Mn2+ + 3H2O

Fe2+

kMnO4

RSM

5 Fe2+ + MnO4- + 8H+

5Fe3+ + Mn2++ 4H2O

Fe3+

kF

Aldehyde

kMnO4

Fe3+ + 3F- Fe F3
RSM

aldehydes+ MnO4- + 4H+

acids + Mn2+ + 2H2O

Suspended solids

AFM

Sulfite,
trio sulfite,

AHM

polythiosulfate
Note: SAM - Sodium Azide Modification
RSM - Rideal Stewast Modification
AFM -Alum Flocculator Modification
AHM- Alkaline - hypochlorite Modification

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

e) Explain how a sample of river water at temperature below or above 20 oC should be
pretreated in preparation for BOD5 test.
Since DO value depends upon temperature. It is a standard for BOD test to carried out at 20 oC
temperature. Firstly, it is a median temperature for all natural water. Secondly, at 20 oC, growth
rate of nitrifying bacteria is limited so that nitrification process does not occur within 5 days.
Therefore, pretreatment of sample should be done to the right temperature.
f) Discuss the effect of seeding and thereby compare two formulas used in calculating
BOD5, ie., the formulas given in Sawyer and McCarty textbook and in Standard
Methods. Do you think two formulas should give the same results?
The effect of seeding may cause the higher value of BOD5 if the oxygen demand of seed itself is
a major part of the oxygen used during incubation. In some cases, BOD value may be reducing
because of lag growth phase of seed.
From Sawyer and McCarty
[ ( DOb - DOi ) ] - [ ( DOb - DOs ) ] ------- (1)
P
= DO value found in blank

BOD (mg/L) =
DOb

DOi = DO value found in dilution sample at the end of incubation period

DOs = DO value found in original undiluted sample
P

= Percent dilution

All unit in mg/L

From Standard Method
BOD (mg/L) =
DO0 =

( DO0 - DOs )
P
DO of diluted sample at initial day 0

DOs =

DO after 5 day incubation at 20oC

------- (2)

In equation (1), when value of DO b equals to DOs the last term can be neglected, and thus the
equation becomes.
BOD (mg/L) =
=

( DOs - DOi )
P
( DO0 - DOs )
P

--------- (3)
--------- (2)

10

Dissolved Oxygen and Biological Oxygen Demand October 30, 2013

From the above, equation (3) is obviously identical to equation (2). In conclusion, both method
will give the same result when DO of blank and DO of undiluted sample are equal which happen
when % dilution is very low.

11

Problem October 30, 2013

Problems (Lab 8 & 9)

1. Which measure, BOD or COD, is more useful is designing and operating a biological
treatment system? Why?
BOD is more useful in designing, but COD is better suited for operation parameter, due to the
following reasons.
In designing, BOD value represents the biodegradable substance which correspond to the
biological processes. While, COD represents chemically oxidizable organic which is normally
higher than BOD. BOD per COD ratio of a sample can give a rough picture of how much
biological oxidizable organic presents in the sample.
In operation, COD test is more widely used, because it requires less time. COD result can be
known within 3 hours, thus it is better. If relationship between BOD and COD is known for a
given wastewater, BOD can be estimated from COD as well.
2. a) If 5-days BOD value of waste is 200 mg/L. Assuming K to be 0.15, what is the
ultimate BOD?
From equation,
=

L (1-10-kt)

BOD at any time t (= 200 mg/L)

Ultimate BOD

Constant Value

(= 0.15)

Time

(=5 days)

So,

Y
Where,

Y
(1-10-kt)

200

243.3 mg/L

(1-10-(0.15x5))
Thus, the Ultimate BOD is

243

mg/L

2. b) Two wastes of equal volume have different BOD values, one with 350 mg/L. and
another 650 mg/L. Draw a graph to show how the oxygen sag curves would differ in the
receiving streams, assuming that stream flows and other conditions are identical.
1

Problem October 30, 2013

The graph represents the DO sag curves, if the stream flows and other conditions are identical,
the waste that has BOD value 650 mg/L can reduce more oxygen in stream water than the less
BOD value. The waste with BOD 650 mg/L also use more time to purify the stream than the
waste with BOD 350 mg/L

DO (mg/L)
Point of discharge

BOD 350 mg/L

BOD 650 mg/L

Distance
3. In the open reflux method for COD determination of sea water sample which has CI concentration about 15 g/L. Mercuric Sulfate must be added give the reasons ? and How
much ?
One of interference in COD test is Cl - which can be oxidized by dichromate as well as other
organic substance and gives Cr3+. Hence, the COD value will be higher than it should be. The
chemical reaction is as follows:
6 Cl- + Cr2O72- + 14 H+ 3Cl2 + 2 Cr3+ + 7 H2O
To prevent this reaction, HgSO4 is added. The mercuric ion can react with chloride ion and form
mercuric chloride complex.
HgSO4 + 2 Cl- HgCl2 + SO42Mercuric sulfate 1 mole react with 2 mole of chloride.
Cl- (2 mole) 71 g
Cl-

reacts with

HgSO4 (1 mole) 296.6 g

15 g/L reacts with HgSO4

= 296.6 15 / 71 = 62.66 g/L

So, 62.66 g/L of mercuric sulfate should be added to eliminate 15 g/L. of chloride ion .
4. What is the theoretical COD of samples containing 300 mg/L of
a) Ethyl alcohol

b) Glucose
2

Problem October 30, 2013

a) Ethyl alcohol reacts with oxygen give the following chemical reaction.
C2H5OH + 3 O2

2 CO2 + 3 H2O

From the reaction, oxygen 3 mole can oxidize 1 mole of C2H5OH.

C2H5OH (1 mole) 46 g

use O2 ( 3 mole) 96 g

C2H5OH

use O2

300 mg/L

= 96 300
46
So, the theoretical COD of sample = 626 mg/L

626 mg/L

b) Glucose reacts with oxygen gives the following equation.

C6H12O6 + 6 O2

6 CO2 + 3 H2O

From reaction, oxygen 6 mole can oxidize 1 mole of C6H12O6.

C6H12O6 (1 mole) 180 g

use O2 ( 6 mole) 192 g

C6H12O6

300 mg/L use O2 = 192 300 = 320 mg/L

180
So, the theoretical COD of sample = 320 mg/L