DO (Dissolved Oxygen)

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 Why Measure Dissolved
Oxygen?

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Scientists want data for…

 Determine the mixing of air and water at

the water’s surface
 Determine what animals can live in the
water

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 Dissolved Oxygen

• Test will measure the amount of free oxygen

“gas” dissolved in your water sample in mg/L
(ppm)
• Dissolved oxygen levels of at least 5 - 6 ppm
(mg/L) are usually required for growth.
• Dissolved oxygen levels of below 3 ppm are
stressful to most aquatic organisms.
Factors affecting the solubility
of Dissolved Oxygen include:

 Temperature
 Atmospheric pressure
 Salinity

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 Biological Influences and
Dissolved Oxygen
 As photosynthesis increases, oxygen levels
increase:

CO2 + H2O Biomass + O2

 As respiration increases due to decay or organic

materials, oxygen levels decrease:

Biomass + O2 CO2 + H2O

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 Taking a Sample and testing

 Rinse sampling bottle 3 times with sample

water
 Submerge bottle in water and allow to fill.
 Tap bottle to release air bubbles
 While bottle is submerged, replace cap
 Ifthere are air bubbles in the bottle, empty and
repeat
 Preserve sample immediately. Test within 2
hours.
 Repeat 3 times. Take the average to see if all
values are within the precision of the kit. 7
Sample Preservation and Sample
Testing
Dissolved Oxygen test kits involve two overall parts:
sample preservation and sample testing.
 Preservation:
 1st - addition of a chemical that precipitates in the
presence of dissolved oxygen
 2nd - addition of a chemical that causes the solids to
dissolve and produce a colored solution. This
should be done in the field.
 Sample Testing:
 Titration of preserved sample. This can be done in
the lab.

Most DO test kits are based on the Winkler

titration method.
 Chemical Reactions

To Preserve DO: Done in the field

O2 + 2 Mn2+ + 2H2O 2MnO2 + 4H+ (pH >10)
Allow precipitate to settle (reaction goes to completion)

2MnO2 + 4H+ + 2I- Mn2+ + I2 (yellow) + 2H2O (low pH)

DO is preserved
--------------------------------------------------------------------------
To Test Sample: This step can be done in the lab.
Na2S2O3 + 4I2 + 5H2O 8I- + 2SO42- + 10H+ + Na+ (the titration)
Starch + I2 blue (to improve endpoint determination)

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DO meter

Cathode O2 + 2H2O + 4e- 4OH-

Anode Pb + 2OH- PbO + H2O + 2e-

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BOD (Biochemical
Oxygen Demand)

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BOD
The oxygen content of water samples stored in
a full, airtight bottle decrease with time
owing to the oxidation of
organic matter by microorganisms

Excretions of aquatic biota, water-soluble
humus compounds and industrial,
domestic and agricultural effluents

BOD is an empirical determination of the
amount of oxygen required to oxidize the
organic matter in the sample
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It can be used to infer the general quality of the water
and its degree of pollution and it is used in:
• Water quality management and assessment,
• Ecology
• Environmental science.

BOD is not an accurate quantitative test and should

be considered as providing an indicator of the quality
of a water body.

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BOD
Determined by incubating the water sample
with aerobic microorganisms under
specific conditions of time and
temperature

BOD5 is based on a 5-day incubating
period at 20oC

BOD = (initial DO – final DO) mg/L
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15
 Biochemical Oxygen Demand:
measurement
 rate at which oxygen is
consumed is expressed
by a rate constant, k
 value of k depends upon
temperature, type of
organic material present,
species of microbes
exerting the BOD
 summary: BOD rate
increases with temp, but
UBOD doesn’t (implies
standardization) 16
BOD
 BOD5 does not represent the total BOD,
since the biological oxidation of organic
matter takes a lot longer than 5 days to
go to completion
 95-99% of the reaction is complete after
20 days
 The BOD test is widely used to
determine the pollution strength of
wastewater and the quality of receiving
surface waters
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Reaction:

CnHaObNc + (n + a/4 - b/2 - 3c/4) O2 ---->

n CO2 + (a/2 -3c/2) H2O + cNH3

2 NH3 + 3O2 ----> 2NO2- + 2H+ + 2H2O

2NO2- + O2 ----> 2NO3-

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Classification of surface water
quality based on BOD values
Degree of pollution BOD5 (mg/L O2)
Very clean < 1.0
Clean 1.1 – 1.9
Moderately polluted 2.0 – 2.9
Polluted 3.0 – 3.9
Very polluted 4.0 – 10.0

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20
BOD - METHODOLOGY
• The water sample is diluted with a suitable
volume of oxygen-saturated water
containing nutrients and incubated in the
dark for 5 days at 20oC
• The DO concentration is determined
before and after incubation and BOD5 is
calculated by difference

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MATERIALS FOR BOD ANALYSIS
o All the reagents required for the DO test
by the Winkler method
o BOD incubation bottles: 250 mL capacity
o Air incubator or water bath at 20  1oC
place in the dark
o Phosphate buffer solution
o Calcium chloride solution
o Magnesium sulfate solution

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MATERIALS FOR BOD ANALYSIS
 Ferric chloride solution
 Dilution water
 HCl 1:1
 NaOH 6M
 pH meter and electrode

NOTE : DISCARD ANY OF THE


REAGENT SOLUTIONS IF THEY SHOW
SIGNS OF BACTERIAL GROWTH
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 SAMPLING & STORAGE FOR
BOD ANALYSIS
 Collect 0.5 – 1 L of sample
 The test should be started as soon as
possible after sampling
 Otherwise, store at 3-5oC in a refrigerator
for up to 24 h.
 Bring the sample to 20oC before starting
the test

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BOD ANALYSIS
 Measure the pH of the sample, and adjust
to pH 7-8 using HCl or NaOH. If the pH is
between 6.5 & 8 do not adjust.

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BOD ANALYSIS
 Completely fill three 250 mL BOD bottles
with the diluted solution, ensuring that no
air bubbles are trapped
 Determine the DO in one bottle
immediately using the Winkler method
 Stopper the other two bottles, water-seal,
place in the air incubator and incubate in
the dark for 5 days at 20oC. After 5 days,
determine the DO using the Winkler
method
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BOD ANALYSIS
 Fill 3 bottles with dilution water in the same way.
Determine DO in one bottle immediately and the other
after incubating for 5 days
 This dilution water blank serves as a rough check on
the quality of the dilution water and it should not
exceed 0.2 mg/L
 Calculate the BOD = F (Di – Df)
Di is the initial DO in the diluted sample
Df is the DO in diluted sample after 5 days of
incubation
F is the dilution factor
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COD (Chemical Oxygen
Demand)

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Chemical oxygen demand

In environmental chemistry,
the chemical oxygen demand (COD) test is
commonly used to indirectly measure the
amount of organic compounds in water.
• Most applications of COD determine the amount
of organic pollutants found in surface water (e.g.
lakes and rivers), making COD a useful measure
of water quality.
• It is expressed in milligrams per liter (mg/L), which
indicates the mass of oxygen consumed per liter
of solution.
• References may express the units as parts per
million (ppm). 29
 The basis for the COD test is that nearly all organic
compounds can be fully oxidized to carbon dioxide with a
strong oxidizing agent under acidic conditions.
The amount of oxygen required to oxidize an organic
compound to carbon dioxide, ammonia, and water is given by:

This expression does not include the oxygen demand caused by the oxidation
of ammonia into nitrate. The process of ammonia being converted into nitrate
is referred to as nitrification. The following is the correct equation for the
oxidation of ammonia into nitrate.

• The second equation should be applied after the first one to include oxidation
due to nitrification if the oxygen demand from nitrification must be known.
• Dichromate does not oxidize ammonia into nitrate, so this nitrification can be
safely ignored in the standard chemical oxygen demand test.
• The International Organization for Standardization describes a standard 30
method for measuring chemical oxygen demand in ISO 6060
• potassium permanganate was not able to effectively oxidize
all organic compounds in water,
• potassium dichromate (K2Cr2O7) has been shown to be the
most effective
The reaction of potassium dichromate with organic compounds is given by:

where d = 2n/3 + a/6 - b/3 - c/2.

Most commonly, a 0.25 N solution of potassium dichromate is
used for COD determination, although for samples with COD
below 50 mg/L, a lower concentration of potassium
dichromate is preferred.
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 BOD determines compounds that can be
biologically oxidized, COD measures
those substances which can be chemically
oxidized.

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METHODOLOGY-COD
o The sample is refluxed with excess
potassium dichromate in concentrated
sulfuric acid for 2 h.
o This oxidizes most of the organic matter in
the sample
o Silver sulfate is included as a catalyst to
speed up the oxidation process

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METHODOLOGY-COD
o After digestion, the unreacted dichromate
remaining in solution is titrated with ferrous
ammonium sulfate:
Cr2O72- + 6Fe2+ + 14H+  2Cr3+ + 6Fe3+ +
7H2O
The amount of dichromate consumed is
calculated and the oxidisable organic
matter is reported in term of oxygen
equivalent
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COD analysis
 COD (mg/L O2) = 8000 x M x (V1 – V2)/Vs
 V is the volume of ferrous ammonium
1
sulfate titrant used to titrate the blank (mL)
 V is the volume of ferrous ammonium
2
sulfate titrant used to titrate the sample
(mL)
 V is the volume of sample (mL)
s
 M is the exact concentration of ferrous ammonium
sulfate titrant (M)

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Problems
 6Cl- + Cr2O72- + 14H+
3Cl2 +2Cr3+ + 7H2O

 Hg2+ + 2Cl- HgCl2

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