Chap 8: Wastewater Sampling and Analysis

Dissolved Oxygen (DO) Analysis

• Surface waters of good quality should be saturated with
dissolved oxygen.
• A fall in DO level is One of the first indications that a water
body is polluted by organic Matter.
• The DO level in water depends on physical, chemical and
biochemical activities prevailing in the water body and, thus, it
is one of the important parameters for assessing the purity of
the water body.
• It is usually determined by Winkler's method", which is based
on the reaction of dissolved oxygen with manganese ions to
form a precipitate of manganese dioxide.
 Biochemical Oxygen Demand (BOD)
The most widely used and accepted measure of biodegradable
organic content of wastewater is the 5-day, 20C BOD value. The
brief analytical procedure is outlined below:
1. Two standard 300-ml BOD bottles are filled completely with the
wastewater of which the BOD is to be measured and the bottles
are sealed.
2. Oxygen content of one bottle is determined immediately.
3. The other bottle is incubated at 20 C for 5 days in total
darkness, after which its oxygen content is measured.
4. The difference between the two DO values is the amount of
oxygen that is consumed by microorganisms during the 5 days
and is reported as the BOD, (5-day BOD) value of the sample.
 Biochemical Oxygen Demand (BOD)

• In practice however, BOD measurements are more complicated

than the simple procedure given above.
• The wastewaters may have a high oxygen demand, high enough
to deplete all the dissolved oxygen in the sample before the end
of 5 days, thus making the test indeterminate.
• So, often the sample is diluted with high purity water to prevent
total depletion of DO.
• The diluting water is made up of deionized water to which
appropriate nutrients, phosphate buffer, trace elements, and seed
organisms (usually settled domestic sewage) are added.
 Biochemical Oxygen Demand (BOD)
A blank is run on the diluting water so that the oxygen demand of
the same material- can be subtracted from the results. The BOD5 is
calculated from the following equation:

BOD 5 in mg / l   D * DOt 0  DOt 5   DOt 0  DOt 5 

sample blank

where D* = dilution factor.

Dilution Procedure

• Two standard 300-ml BOD bottles are taken for this purpose

• In one bottle deionized water with appropriate nutrients,

phosphate buffer, trace elements and seed organisms are added

• In second bottle the required amount of wastewater along with

deionized water with appropriate nutrients, phosphate buffer,
trace elements and seed organisms are added.

• Both the bottles are incubated at 20 C for 5 days in total

darkness, after which its oxygen content is measured.

• The required amount of wastewater is calculated based on

certain assumptions
Dilution Procedure sample calculation

For example, Lets assume that the BOD of the ww is 200 mg/l and
5 mg/l of DO is consumed in five days
So the sample required will be 5/200 = 0.025 or 2.5 percentage
In a standard bottle of 300 ml and the sample size would be
300 x 0.025 = 7.5 ml
Validation of BOD test

• At least 2 mg/l of oxygen should remain after incubation

• Minimum 1 mg/l of dissolved oxygen should be used during the
incubation period

volumeofdilutedsample
DilutionFactor 
volumeofundilutedsample

volumeofundilutedsample
SampleSize%  100
volumeofdilutedsample
 BOD Test

• When the standard 5-day/20'C conditions are used,

approximately two-thirds of the carbonaceous material is
broken down;

• an incubation of about 20 days is needed for nearly complete

breakdown.

• Nitrogenous nutrients can create problems in-the BOD test.

• Because of the slowness of the nitrification process, the

oxygen demand of the nitrifying bacteria is assumed to be
negligible in the standard 5-day incubation period at 20C.
 BOD Test
• The actual environmental conditions of temperature, biological
population, oxygen concentration, etc., are impossible to
reproduce in the test; hence, care must be exercised in
extrapolating the test results to the actual stream oxygen
demands.

• In addition, many industrial wastewaters contain toxic

materials which interfere with the growth of microorganisms
thus making the BOD test unreliable or even inapplicable
without the addition of suitable inoculum.
 BOD Test
• Similarly, the presence of algae in the wastewater affects the
BOD test by leading to higher BOD values even when the test
is performed in darkness.
 Chemical Oxygen Demand (COD)

• In the COD test, the oxidizing bacteria of the BOD test are
replaced by a strong oxidizing agent under acidic conditions.

• A sample of the waste- water containing organic material is

mixed with an excess of potassium dichromate and sulphuric
acid and the mixture is heated under total reflux conditions for
a period of 2 hours.
 Chemical Oxygen Demand (COD)
• During digestion, the chemically oxidizable organic material
reduces a stoichiometrically equivalent amount of dichromate,
the remaining dichromate is titrated with standard ferrous
ammonium sulphate' solution.

• The amount of potassium dichromate reduced gives a measure

of the amount of oxidizable organic material.

• Dichromate has advantages over other oxidants in oxidizing

power and applicability to a wide variety of samples.
• The COD test does not distinguish between organic
materials that are biodegradable and those that are not, and,
hence, gives a measure of the total oxidizable organic
material in the sample.
• Due to this, the COD test results are higher than those of
BOD tests carried out on the same sample.
• If inorganic substances such as chlorides and nitrites are
present in the waste- water, they interfere with the COD test
since they are also oxidized by dichromate and create an
inorganic COD that leads to an error in the measurement.
• Chloride interference can be eliminated by adding mercuric
sulphate to the sample prior to the addition of other reagents,
and nitrite interference can be overcome by adding
sulphamic acid to the di-chromate solution.
• The COD test-is much more useful than the BOD test for
estimating the oxygen requirements of certain industrial
wastewaters.
• It is valuable for wastes where BOD test is not applicable due
to the presence of toxic substances, low rate of oxidation, or
other similar factors.
• Ratios of BOD to COD can be employed to get an indication
of the degree of biotreatability of the waste.
• Ratios of 0.8 or higher indicate wastes that are-highly
amenable to biological treatment, while lower ratios indicate
that the wastes are not favorable to biological treatment.
Total Organic Carbon (TOC)
• Total organic carbon test is based - on the oxidation of the
carbon of the organic matter to carbon dioxide, which is
measured by non-dispersive infrared analyzer.
• Alternatively, the carbon dioxide can be' reduced to
methane, which is then measured by a flame ionization
detector.
• In this test, a few micro-litres (5 to 10 ) of the aqueous
sample are injected into a combustion tube containing a
catalyst and heated to 900C in a constant flow of air.
• Water is vaporized and the carbonaceous matter is
oxidized to CO2. and steam. Outside the combustion tube,
the steam is condensed and removed.
• CO2 is swept into a non-dispersive infrared analyzer,
which measures the amount of CO2.
• The concentration of CO2 is directly proportional to the
concentration of total carbon present in the original sample
and it includes both organic and inorganic carbon.

• Inorganic carbon can be measured separately using an acid

catalyst at 150C, which is below the temperature at which
organic matter is oxidized.

• Organic carbon content can then be obtained by subtracting

the inorganic carbon from the total result.

• A flow diagram for a dual combustion tube total carbon

analyzer is shown in Fig. 8.1.
• The TOC test can be performed in a relatively short period of
time (few minutes) compared to BOD and COD measurements
and, hence offers a valuable supplement to BOD and COD
estimations.

• Because of this advantage, an empirical correlation between

TOC and COD or BOD, that is specific to a particular plant
operation, can be established and from this it is possible to
obtain a tentative estimate of the plant performance quickly.
 Carbon Types
• Total Carbon (TC) – all the carbon in the sample, including
both inorganic and organic carbon

• Total Inorganic Carbon (TIC) – often referred to as inorganic

carbon (IC), carbonate, bicarbonate, and dissolved carbon
dioxide (CO2).

• Total Organic Carbon (TOC) – material derived from

decaying vegetation, bacterial growth, and metabolic activities
of living organisms or chemicals.