Joshua R.

Masapequena November 7, 2018

BS Biology I Chem 23. 1 Sec. 4

Post Lab Report

Experiment 5
Dissolved Oxygen by the Winkler Method (Winkler Titration)

I. Introduction
Dissolved oxygen refers to the level of free, non-compound oxygen present in water or
other liquids. It is an important parameter in assessing water quality because of its influence on
the organisms living within a body of water. Dissolved oxygen enters water through the air or as
a plant byproduct. From the air, oxygen can slowly diffuse across the water’s surface from the
surrounding atmosphere, or be mixed in quickly through aeration, whether natural or man-made.
The aeration of water can be caused by wind (creating waves), rapids, waterfalls, ground water
discharge or other forms of running water. A dissolved oxygen level that is too high or too low
can harm aquatic life and affect water quality. Non-compound oxygen, or free oxygen (O2), is
oxygen that is not bonded to any other element. Dissolved oxygen is the presence of these free O2
molecules within water. The bonded oxygen molecule in water (H2O) is in a compound and does
not count toward dissolved oxygen levels. One can imagine that free oxygen molecules dissolve
in water much the way salt or sugar does when it is stirred.
Dissolved oxygen is necessary to many forms of life including fish, invertebrates, bacteria
and plants. These organisms use oxygen in respiration, similar to organisms on land. Fish and
crustaceans obtain oxygen for respiration through their gills, while plant life and phytoplankton
require dissolved oxygen for respiration when there is no light for photosynthesis. The amount of
dissolved oxygen needed varies from creature to creature. Bottom feeders, crabs, oysters and
worms need minimal amounts of oxygen (1-6 mg/L), while shallow water fish need higher levels
(4-15 mg/L). Microbes such as bacteria and fungi also require dissolved oxygen. These organisms
use DO to decompose organic material at the bottom of a body of water. Microbial decomposition
is an important contributor to nutrient recycling. However, if there is an excess of decaying organic
material (from dying algae and other organisms), in a body of water with infrequent or no turnover
(also known as stratification), the oxygen at lower water levels will get used up quicker.
The Winkler Method is a technique used to measure dissolved oxygen in freshwater
systems. 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. This test
is performed on-site, as delays between sample collection and testing may result in an alteration
in oxygen content. The Winkler Method uses titration to determine dissolved oxygen in the water
sample. A sample bottle is filled completely with water (no air is left to skew the results). The
dissolved oxygen in the sample is then "fixed" by adding a series of reagents that form an acid
compound that is then titrated with a neutralizing compound that results in a color change. The
point of color change is called the "endpoint," which coincides with the dissolved oxygen
concentration in the sample. Dissolved oxygen analysis is best done in the field, as the sample will
be less altered by atmospheric equilibration.
The concentration of dissolved oxygen can be readily, and accurately, measured by the
method originally developed by Winkler in 1888. Dissolved oxygen can also be determined with
precision using oxygen sensitive electrodes. 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.
The concentration of dissolved oxygen (DO) is one of the most important indicators of the
overall health of a body of water. Waters with consistently high levels of DO (> 6 mg/L) typically
support the most diverse biological communities. Waters with consistently low DO levels (< 3
mg/L) may be virtually devoid of aquatic life or may harbor only a few species adapted to such
conditions.

II. Methodology
For the collection of samples, seawater were collected using standard (3) DO bottles. These
DO bottles were rinsed twice with the sample being analyzed. If a rubber tubing is used to transfer
sample to DO bottle, always keep the tube beneath the surface of the water as the bottle is filled.
The water collected was allowed to overflow from the top of the DO bottle and stopper at once
and that no air bubble should be entrapped in the bottle.

For the reagents utilized in the experiment, there were 6 reagents needed. These reagents were
prepared according to the procedures provided. (to be continued…)

In standardizing Thiosulfate, 2.5 g KI was dissolved in an Erlenmeyer flask with

approximately 100 mL distilled water. 12.5 mL of 1+9 H2SO4 in the solution was also added. To
obtain 25 mL of 0.025 N K2Cr2O7, a volumetric pipette was used and was transferred to the
Erlenmeyer flask and was mix thoroughly afterwards. Titrate with 0.025 N Na2S2O3 until pale
straw yellow was obtained. 1-2 mL of starch indicator was added and titration was continued with
the remaining liberated I2 with 0.025 N Na2S2O3 until the first disappearance of the blue color.
Lastly, the normality of sodium thiosulfate was calculated.

In determining the DO of the samples, each samples were added with 2 mL manganese
sulfate solution as well as 2 mL alkali-iodide-azide reagent. The flasks were mixed by inverting it
several times (Note: Stopper with care to exclude air bubbles). Then, three samples were left to
stand until precipitate has settled at least 1/3 of the way down the bottle. 2 ml of concentrated
H2SO4 was added carefully by allowing acid to run down the neck of the bottle. It was thoroughly
mixed and just like the previous mixing, no air should be trapped in the bottle. 100 ml of each
samples were transferred into 250 mL Erlenmeyer flask and was titrated with thiosulfate solution
to a pale straw color. 1 mL starch solution was added and titration was continued until the first
disappearance of blue color. The volume of titrant, ppm of D.O., and other essential data were
recorded.

III. Results
A. Standardization of Sodium Thiosulfate solution
Primary Standard used:
Formula mass of 1O standard:
% Purity of 1O standard:
Trials 1 2 3
Weight of K2Cr2O7 (g)

Final Reading Na2S2O3 (ml)

Initial Reading Na2S2O3 (ml)
Volume Na2S2O3 (ml)

Molarity of Na2S2O3
Ave. Molarity of Na2S2O3

B. Sample Analysis
Source of Water sample:
Trials 1 2 3
Volume of sample use for
titration (ml)
Final Volume Na2S2O3 (ml)
Initial Volume Na2S2O3 (ml)
Net Volume Na2S2O3 (ml)

mg O2 in sample
% O2 in sample
Average % O2 in sample
IV. Discussion

V. Summary and Conclusion

VI. References/Literature cited