CHEM181: Chemistry for Engineers - Laboratory Determination of Dissolved Oxygen Content of Water

CHEM181: CHEMISTRY FOR ENGINEERS - LABORATORY

Name Course & Year

Section Date

Experiment No. 8
DETERMINATION OF DISSOLVED OXYGEN CONTENT OF WATER

Objective: To determine dissolved oxygen (DO) in the given water sample

Apparatus: 3 - 300mL glass stoppered BOD bottle acid burette

3 - 500mL Erlenmeyer flask burette clamp
1 - 100mL graduated cylinder pipettor
1 - 10mL graduated cylinder iron stand
3 - 2mL pipettes wash bottle

Materials: 6mL manganous sulfate solution 6-mL alkali-iodide-azide solution

6mL concentrated sulfuric acid solution 0.025N sodium thiosulfate solution
6mL starch solution 3 sets 300-mL water sample

Theory:
Dissolved oxygen (DO) determination measures the amount of dissolved (or free) oxygen
present in a volume of water or wastewater. Dissolved oxygen is used as an indicator of the
health of a water body where higher dissolved oxygen concentrations are correlated with high
productivity and little pollution. It is a measure of the ability of water to sustain aquatic life. Aerobic
bacteria and aquatic life such as fish need dissolved oxygen to survive. If more oxygen is
consumed than is produced, dissolved oxygen levels decline and some sensitive animals may
move away, weaken or die. DO is determined by the titrimetric method developed by Lajos
Winkler called the Winkler Method. This technique is used to measure dissolved oxygen in
freshwater systems. This test is performed on-site as delays between sample collection and
testing may result in an alteration in oxygen content.
CHEM181: Chemistry for Engineers - Laboratory Determination of Dissolved Oxygen Content of Water

Procedure:
Preparation of the Sample
1. Fill three 300mL BOD bottles up to the brim. Cover the bottle and remove excess water.
Make sure no bubbles are formed.
2. Immediately add 2mL manganous sulfate solution to the collection bottle by inserting a
calibrated pipette just below the surface of the water sample in each BOD bottle. Squeeze
the pipette slowly so no bubbles are introduced via the pipette. (If the reagent is added above
the sample surface, you will introduce oxygen into the sample.)
3. Add 2 mL of alkali-iodide-azide reagent in the same manner in each BOD bottle.
4. Stopper the BOD bottle with care to be sure no air is introduced.
5. Mix the sample by inverting several times. Check for air bubbles. Discard the sample and
start over if any are seen.
If oxygen is present, a brownish-orange cloud of precipitate or floc will appear.
6. When this floc has settled to the bottom, mix the sample by turning it upside down several
times and let it settle again.
7. Add 2 mL of concentrated sulfuric acid using a pipette held just above the surface of the
sample.
8. Carefully insert the stopper and invert several times to dissolve the floc.
9. When the floc is dissolved, the sample is "fixed”. (DO determination can be measured
immediately or after 8 hours.)
Titration
10. Prepare the titration set-up. Rinse the burette with 0.025N sodium thiosulfate. After rinsing,
fill the burette with 0.025 N sodium thiosulfate solution.
11. Take the initial reading of the volume (at most 2 decimal places) of sodium thiosulfate inside
the burette.
On other glassware like beakers, flasks, and graduated cylinders, the zero mark is at the
bottom of the scale, with values increasing going up the glassware. A burette, however, has
zero at the top with values increasing going down the glassware.

Figure 1. Reading the volume from a burette

Source: http://faculty.sdmiramar.edu/fgarces/LabMatters/ChemTech/modules/buret/PIC/47102ST.JPG
CHEM181: Chemistry for Engineers - Laboratory Determination of Dissolved Oxygen Content of Water

12. Measure 203mL of the prepared sample in the BOD bottle and transfer to a 500mL
Erlenmeyer flask.
13. Add 2 mL starch solution. The sample will turn blue.
14. Titrate the sample slowly with 0.025N Na2S2O3 until the end point is reached. The end point
is reached when the blue color of the sample disappears from the last drop of the titrant.
NOTE: During titration, be especially careful that each drop is fully mixed into the sample
before adding the next drop. After adding each drop of the titrant, swirl the
Erlenmeyer flask containing the sample.
15. Record the final reading of the volume (at most 2 decimal places) of sodium thiosulfate inside
the burette.
16. The concentration of dissolved oxygen in the sample is equivalent to the number of milliliters
of titrant used. Each mL of sodium thiosulfate added is equivalent to 1 mg/L dissolved
oxygen.
17. Repeat the titration for another 2 trials.

Figure 8. The Flow Diagram for DO Experiments

Source: https://www.quora.com/Why-is-starch-indicator-important-in-a-dissolved-oxygen-analysis
CHEM181: Chemistry for Engineers - Laboratory Determination of Dissolved Oxygen Content of Water

Chemical Reactions:

1. water sample + manganous sulfate solution + alkali-iodide azide reagent

2Mn 2 + (aq) + 4OH − (aq) → 2MnO 2( s) + 2H2 O (l)

• procedure involves the oxidation of Mn 2 + and reduction of O 2

• If O 2 is present, a brownish-orange formation of MnO2 occur which stabilizes and locks
up the dissolved O2 prior to analysis.

2. addition of concentrated sulfuric acid

MnO 2(s) + 2I− (aq) + 4H+ (aq) → Mn 2 + (aq) + I2(aq) + 2H2O(l)

• solution turns to pale yellow
• procedure involves oxidation of iodide ion and reduction of Mn 4 +
• The I2 formed is directly related to the amount of DO in the sample which is determined
by titrating it with Na2S2O3 and using starch solution as indicator.

3. titration with Na2S2O3

2S 2O3 2 − (aq) + I2(aq) → S 4 O 6 2 − (aq) + 2I− (aq)

• procedure involves oxidation of S 2 O 3 2 − and reduction of I2

Results:

Volume of Burette reading in mL Volume of Dissolved

Trial sample in Na2S2O3 used Oxygen
mL Initial reading Final reading in mL in g/mL
1

Volume of Na2S2O3 used = final reading – initial reading

Computations:
CHEM181: Chemistry for Engineers - Laboratory Determination of Dissolved Oxygen Content of Water

Questions:
1. Where does dissolved oxygen come from?

2. Explain dissolved oxygen saturation.

4. What are the factors that affect the dissolved oxygen content in water? Explain how they
affect DO content.

5. Give 5 examples of freshwater organisms and their dissolved oxygen requirements.

6. Give 5 examples of saltwater organisms and their dissolved oxygen requirements.