LJIAS

Department of Biotechnology and Biochemistry

Semester 6

Aim: Determination of Chemical Oxidation Demand.

Introduction:
Pollution of water by organic and inorganic chemicals is of serious environmental concern. Since
all natural water waste contain bacteria and nutrients, almost any waste compound introduced
into such water, bacteria and nutrients will initiate biochemical reactions. The organic content of
wastewater is traditionally measured using lumped parameters such as Biochemical Oxygen
Demand (BOD), Chemical Oxygen Demand COD), total suspended solids (TSS).

The estimation of COD is of great importance for waters having unfavorable conditions for the
growth of microorganisms, such as presence of toxic chemicals. In such waters, BOD cannot be
determined accurately. Chemical Oxygen Demand determines the amount of oxygen required to
chemically oxidize all soluble and particulate organic substances in water using oxidizing agents
such as potassium dichromate under specific reflux conditions. It is a significant parameter to
determinate water characteristic. This test is widely used to determine the degree of pollution in
water bodies and their self-purification capacity, efficiency of treatment plants, pollution loads,
and provides rough idea of Biochemical Oxygen Demand (BOD) which can be used to determine
sample volume for BOD estimation.

However, the limitation of the test lies in its inability to differentiate between the biologically
oxidizable and biologically inert material and to find out the system rate constant of aerobic
biological stabilization. Most of the organic matters are-destroyed when boiled with a mixture of
potassium dichromate and sulphuric acid producing carbon dioxide and water. The COD value
depends on oxidizing agent, pH, temperature, and period of time. It is expressed as ppm or
milligrams per liter (mg/L) of oxygen. COD is always higher than BOD, by approximately 2 to 3
times of BOD.

There are several methods available for the determination of Chemical Oxygen Demand such as
Open Reflux Method, Closed Reflux - Titrimetric Method, Closed Reflux - Spectrophotometeric
method, but the wet chemical method (dichromate method) is the most common method used
because unlike other methods it doesn't require to have a clear unclouded measurement solution.

Principle:
Most types of organic matter are oxidized by a boiling mixture of chromic and sulfuric acids. A
sample is refluxed in COD digestor under strongly acidic conditions for two hours, at 150oC,
with a known volume of potassium dichromate (K2Cr2O7), which is a strong oxidizing agent.
Oxidisable organic compounds react with K2Cr2O7, reducing the dichromate ion (Cr2O72-) to a
green chromic ion (Cr3+). After digestion, the remaining unreduced K2Cr2O7 is titrated with
ferrous ammonium sulfate using Ferroin indicator to determine the amount of K2Cr2O7 consumed
and this gives us the oxidizable organic matter in terms of oxygen equivalent.

Determination of COD Page 1

The COD digestion solution contains mercuric sulphate and silver sulphate. Mercury is used to
complex interferences from chloride and silver acts a catalyst to promote oxidation of certain
organic compounds such as linear aliphatic compounds, aromatic compounds and pyridine
during the digestion step.

Ferroin indicator is used to indicate change in oxidation-reduction potential of the solution and
indicates the condition when all dichromate has been reduced by ferrous ion. It gives a very
sharp brown color change.

Chemical Reaction:

The reactions taking place during the oxidation can be expressed by the following series of
equations.
Oxidation reaction: Cr2O72- + 8H+ → 2Cr3+ + 4H2O + 3O
Iron (II)/dichromate titration: 6Fe2+ + Cr2O72- + 14H+ → 6Fe3+ + 2Cr3+ + 7H2O
Chloride interference reactions: Cr2 O7 2- +6Cl- + 14H+ → 2Cr3+ + 3Cl2 + 7H2 O
Hg2+ + 4Cl- → [HgCl4]2-
Cr3+ + 6Cl- → [CrCl6]3-
Ag+ + Cl- → AgCl

Interference:
Chloride is the primary interference when determining the COD level of a sample. Chloride
reacts with silver ions, present within the COD digestion solution, to precipitate silver chloride
thus inhibiting the catalytic activity of silver. Bromide, iodide and any other reagent that
inactivates these silver ions can lead to similar interferences. The difficulties caused by the
presence of chloride can be overcome largely, though not completely, by complexing with
Mercuric Sulphate (HgSO4).

Requirements:
1. Distilled water 9. Sulfuric acid reagent (Conc. H2SO4
2. Porcelain pieces containing Ag2SO4)
3. Standard Potassium Dichromate 10. COD digestor
solution K2Cr2O7 (0.25 N) 11. Waste water sample
4. Standard Ferrous Ammonium 12. Conical flasks
Sulfate Fe(NH4)2SO4 (0.1 M) 13. Pipettes
5. Mercuric sulfate (Powdered HgSO4) 14. Burette
6. Phenanthroline Ferrous Sulfate 15. Burette stand
(Ferroin) indicator solution 16. Refluxing tube
7. Concentrated Sulphuric acid (H2SO4 17. Condensor tubes
sp. gr. 1.84)
8. Silver sulfate (Ag2SO4)

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Procedure:
1. Take a clean Refluxing tube and add 400 mg of mercuric sulphate.
2. Add 10 mL of potassium dichromate by pipette.
3. Add 3-4 porcelain pieces into the refluxing tube.
4. Then add 30 mL of conc. sulphuric acid reagent containing AgSO4 by measuring
cylinder. Acid should be added in controlled manner with proper mixing.
5. Then add 20 mL of sample in the Refluxing tube. If the sample color changes to green,
dilute the sample and repeat the procedure for diluted sample.
6. Connect the Refluxing tube through the condenser and reflux for a minimum period of 2
hours at 150°C ± 2°C in a preheated COD digester.
7. Cool Refluxing tube to room temperature and add 60 mL distilled water through
condenser. After 5-10 minutes detach the condensor tube.
8. Carefully transfer the mixture into 250 mL Erlenmeyer flask.
9. Then titrate the excess dichromate with 0.1 M Ferrous Ammonium Sulfate, by using 2 to
4 drops of ferroin indicator. At the end point the color turns from blue-green to reddish
brown and note down the burette reading as "B" mL.
10. A blank sample of distilled water is refluxed through the same COD testing procedure as
the waste water sample. Again note down the amount of FAS consumed as "A" mL until
the color changes from blue-green to reddish brown. The purpose of running a blank is to
compensate for any error that may result because of the presence extraneous organic
matter in the reagents.
11. Calculate the COD (mg O2/L) as follows:
𝑚𝑔 𝑂2 (𝐴 − 𝐵) × 𝑀 × 8000
𝐶𝑂𝐷 ( )=
𝐿 𝑣𝑜𝑙 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒(𝑚𝐿)

where:

A = mL FAS used for blank,

B = mL FAS used for sample,
M = molarity of FAS, and
8000 = milliequivalent weight of oxygen (8) X 1000 mL/L.

Permissible limit:

According to CPCB, the permissible limit for COD is 250 mg O2/L.

Determination of COD Page 3

Observation Table:

Sr. Volume of Volume of titrant FAS

Sample
No. sample(mL) consumed (mL)
Blank
1. (Distilled 20 mL mL
water)
2. Sample 20 mL mL

Burette Solution: 0.1 M Ferrous Ammonium Sulphate

Indicator: Ferroin indicator
End Point: Appearance of brown color

Calculations:
Volume of Ferrous Ammonium Sulphate for Blank (A) = mL
Volume of Ferrous Ammonium Sulphate for Sample (B) = mL
Molarity of Ferrous Ammonium Sulphate = 0.1 M
Volume of sample = 20 mL

We know,

𝑚𝑔 𝑂2 (𝐴 − 𝐵) × 𝑀 × 8000
𝐶𝑂𝐷 ( )=
𝐿 𝑣𝑜𝑙 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒(𝑚𝑙)

Putting the values in the above formulae, we get

= mg O2/L or ppm

Results:
The COD content of the given waste water sample is mg O2/L or ppm

Conclusion:
The determination of COD is quick and accurate method for the analysis of oxidisable organic
matter present in the waste water sample.

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