Experiment 8 (BOD) - Lab Manual

Analytical Chemistry 1 Laboratory Manual
For Engineering students
EXPERIMENT 6
Determination of Biological Oxygen Demand using the Modified Winkler
Method
Knowledge of the dissolved oxygen (O2) concentration in seawater is often necessary in

environmental and marine science. It may be used by physical oceanographers to study
water masses in the ocean. It provides the marine biologist with a means of measuring
primary production - particularly in laboratory cultures. For the marine chemist, it provides
a measure of the redox potential of the water column.
The concentration of dissolved oxygen (DO) can be readily, and accurately, measured by
the method originally developed by Winkler in 1888 (Ber. Deutsch Chem. Gos., 21, 2843).
DO can also be determined with precision using oxygen sensitive electrodes; such
electrodes require frequent standardization with waters containing known
concentrations of oxygen. They are particularly useful in polluted waters where oxygen
concentrations may be quite high. In addition, their sensitivity can be exploited in
environments with rapidly-changing oxygen concentrations. However, electrodes are
less reliable when oxygen concentrations are very low. For these reasons, the Winkler
titration is often employed for accurate determination of oxygen concentrations in
aqueous samples. Constant water monitoring of effluents as well as raw and treated water
also include testing for dissolved oxygen. Since an adequate supply of DO is required to
support life in a given body of water, a determination of the amount of DO provides a
means of assessing the quality of water with respect to sustaining life. The table below
provides a general guideline for interpretation of DO readings:
Table 1. General interpretation of DO readings
0-2 ppm Not enough oxygen to

support life
2-4 ppm Only few kinds of fish
and insects can
survive
4-7 ppm Acceptable for warm
water fish
7-11 ppm Very good for most
stream fish including
cold water fish
The biological oxygen demand (BOD) test is an empirical bioassay-type procedure which
measures the dissolved oxygen consumed by microbial life while assimilating and
oxidizing organic matter present. The standard test conditions include dark incubation at
20°C for a specified time period (often 5 days). DO is measured initially and after
incubation, and the BOD is computed from the difference between initial and final DO.
41 | P a g e
In this experiment, the DO and 5-day BOD values for a real water sample will be analyzed
using the modified Winkler method.
CHEMICALS/REAGENTS:
Distilled water
MnSO4·2H2O crystals
NaOH pellets
KI crystals
Na2S2O3·5H2O
Concentrated H2SO4
Starch
KIO3
MATERIALS/EQUIPMENT:
Volumetric flasks, 50-mL, 100-mL, 250-mL, & 500-mL
Erlenmeyer flasks, 250-mL
Burette, 25-mL
Beaker, 250-mL
Syringes, 50-mL
PROCEDURE:
A. Preparation of reagents
 MnSO4 Solution
Dissolve 20.0 g of MnSO4·2H2O crystals in about 30 mL distilled water. Filter resulting
solution into 50-mL volumetric flask if needed, then dilute to mark.
 Alkaline KI solution
Dissolve 18 g NaOH pellets and 10 g KI in 30 mL distilled water in a small beaker
submerged in a water bath. Transfer into a 50-mL volumetric flask and dilute to mark.
 0.125 M Na2S2O3 stock solution
Weigh about 7.8 g of Na2S2O3·5H2O in a 250-mL beaker and dissolve with 100 mL pre-
boiled distilled water. Transfer into 250-mL volumetric flask, wash beaker with pre-boiled
distilled water and combine washings into volumetric flask. Dilute to mark.
 0.5 M H2SO4 solution
Dilute 6.94 mL of concentrated H2SO4 in 500 mL volumetric flask. Dilute to mark.
 1% starch solution
Moisten 0.50 g of soluble starch with a small amount of distilled water until a smooth paste
is obtained. Pour slowly into 100 mL boiling water. Mix thoroughly until dissolved. Cool to
room temperature.
 0.0125 M Na2S2O3 standard solution
42 | P a g e
Prepare approximately 0.0125 M Na2S2O3 in 500-mL volumetric flask.
B. Standardization of Na2S2O3 solution

Weigh 0.15 g of KIO3 to the nearest 0.1 mg and dissolve in 50 mL of pre-boiled distilled
water. Transfer into a 100-mL volumetric flask and dilute to mark. Mix thoroughly. From
the prepared KIO3 solution, transfer 10.00-mL aliquots into three 250-mL Erlenmeyer
flasks. Add about 20 mL distilled water into the flask. Add about 1 g KI crystals and 10
mL of 0.5 M H2SO4. Add reagents one at a time. Swirl to mix and immediately titrate with
standard thiosulfate solution with constant swirling. When solution becomes pale yellow,
add 1 mL of the starch solution and continue titration until the disappearance of the blue
color. Repeat using the 2nd and 3rd aliquots. Calculate the molarity of standard Na2S2O3.
C. Analysis of unknown sample

Obtain four 50-mL syringe (fresh from the pack) and label as Day1A, Day1B, Day5A
and Day5B, respectively. Carefully and slowly, draw the water sample into the four
syringes. Care must be observed as not to allow air bubbles to be trapped inside the
syringe. [Ideal sampling depth is 6 feet from the surface for bodies of water]. Dry the tips
of the syringe with tissue. Carefully expel sample until the plunger is at exactly at the 30
mL mark. Seal the tip of the syringes labeled Day5A and Day5B with paraffin film and
store in dark to be processed after 5 days.
For the Day1A and Day1B samples carefully draw MnSO4 solution until the 35 mL
mark. Be careful not to overshoot the mark and that no air bubbles are trapped inside the
syringe. Mix slowly by cautiously tipping the syringe repeatedly. Draw the alkaline iodide
solution up to the 40 mL mark and mix slowly as above. Finally, draw in the H 2SO4 reagent
up to the 50 mL mark and mix well until all brownish yellow particles dissolve.
Expel the contents of the syringe in separate 250-mL Erlenmeyer flasks. Rinse the
inside of the syringe with distilled water and add the washings. Add about 5 mL of starch
solution (freshly prepared) into the flasks. Titrate the contents of the flask with the
standardized Na2S2O3 titrant for the D.O. determination. Calculate D.O. in ppm.
Do the same with the Day5A and Day5B samples after the 5-day incubation period.
GUIDE QUESTIONS:
1. What are the pertinent chemical equations in the standardization of sodium
thiosulfate solution?
2. Explain the reason for the addition of H2SO4 and excess KI.
3. What is the effect of adding the acid before KI?
4. Present the pertinent chemical equations involved in the analysis.
5. Explain the stepwise production of titratable I2 from dissolved O2 in the sample.
6. Explain why the reagents are added in the specific sequence described in the
experiment.
43 | P a g e
7. Explain the would-be effects (if any) of the following considerations:

a. Water sample was made to stand overnight before analysis
b. MnSO4 was added and the solution was made to stand overnight before the
analysis
8. What are the possible sources of errors and their effect on the calculated
parameters?
44 | P a g e

Experiment 8 (BOD) - Lab Manual

Uploaded by

Copyright:

Available Formats

Experiment 8 (BOD) - Lab Manual

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Experiment 8 (BOD) - Lab Manual

Uploaded by

Copyright:

Available Formats

What is the Winkler method and what is it used to measure?

What factors affect the reliability of oxygen electrodes and why is the Winkler method often preferred?

Analytical Chemistry 1 Laboratory Manual

For Engineering students

Knowledge of the dissolved oxygen (O2) concentration in seawater is often necessary in

0-2 ppm Not enough oxygen to

Prepare approximately 0.0125 M Na2S2O3 in 500-mL volumetric flask.

B. Standardization of Na2S2O3 solution

C. Analysis of unknown sample

7. Explain the would-be effects (if any) of the following considerations:

You might also like