R. J. C. De Guzman / Chemistry 26.

1 (2017) Page |1

Quantitative Determination of Dissolved Oxygen Content by Winkler Redox

Titration
Raphael Joshua C. De Guzmana
a
Institute of Biology, University of the Philippines Diliman
Performed 17 March 2017; Submitted 29 March 2017

ABSTRACT

The Winkler Titration was used to measure the dissolved oxygen (DO) present in the creek in the Science
Complex Park. The titration is dependent on the oxidative nature of DO which allows to liberate Mn(III)
ions from Mn(II) ions and thus forms Mn(OH) 3 which has strong alkali. When added with I, oxidized Mn
(III) ions is reduced back to Mn (II) ions and also liberates I 2 ions which is equivalent to the DO content in
the water sample. The titrant used was thiosulfate (Na 2S2O3) and the analyte was iodine (I 2). From the
calculations performed after the experiment (See appendix), Thiosulfate was standardized at
0.0011298307 M and the DO content of the water sample is at 2.049 ppm which theoretically cannot
support life. Relative standard deviation is at 41.595 ppt and its confidence interval at 95% is
2.048631741± 0.211547453 .
_________________________________________________________________________________________________________________________
Introduction life
0-2 Highly polluted Cannot support
life
Dissolved oxygen (DO) measures the amount of
free, non-compound oxygen ions in natural
Methodology
water and wastewater depending on the
physical, chemical, and biochemical processes
Water sample to be tested came from the creek
involved. Determination of dissolved oxygen is
in the Science Complex Park near the Institute of
important for marine biologist and water
Mathematics building. The water was collected
chemist to study water masses or as means of
and stored in a 400 mL glass bottle, made sure
measuring water quality to sustain life (see
that the sample has no air trapped inside, and
Table 1). The Winkler Redox Titration, also
then immediately covered with aluminium foil
known as Iodometric Titration, is the commonly
to prevent exposure to sunlight because sunlight
used way of determining DO of water sample
increases dissolved oxygen via free radical
(Skoog, 2015). The titration is different from an
reaction. (Fondriest Environmental Ink, 2014)
Iodimetric titration which uses Na2S2O3 as
The sample was stored inside the lockers for
analyte and I2 as titrant (Fondriest
two days therefore the DO content of the sample
Environmental, Inc, 2014). This method was
decreased since heterotrophic organisms that
developed by Lajos Winkler in 1888 and is
existed inside the sample consumed oxygen for
based on oxidative property of DO. (JM Soden,
respiration.
n.d)
The Winkler method utilized the following
The oxidative nature of DO enables to liberate chemicals namely: Manganese Sulfate
Mn(III) ions from Mn(II) ions and thus forms (MnSO4*2H2O), which was the source of Mn
Mn(OH)3 which has strong alkali. When added ions; KI, the source of Iodide ions; KIO 3, the
with I, oxidized Mn (III) ions is reduced back to primary standard; H3PO4,;NaOH;
Mn (II) ions and also liberates I2 ions which is Na3N,;Na2S2O3*5H2O,; the source of thiosulfate
equivalent to the DO content in the water ions. The experiment made use of 50 mL
sample. From here, the DO content can be burettes, 50-,100-mL beakers, watch glass, 1-mL
measured using parts per million. (Clemente, syringe, 250 mL Erlenmeyer flask, 250 mL
2013) volumetric flask, Analytical balance, Top-loading
balance, and hot plate.
Relationship between DO Content and Water
Quality (Table 1)
DO Content Water Quality Life Support Solution preparation was done during the first
(ppm), 20°C day of the experiment. These include 25 mL 4.0
8-9 Clean good Stream fishes M MnSO4 from MnSO4*2H2O crystals, which the
water calculated and weighed amount (18.7 g) was
6.7-7.9 Slightly polluted Cold water fishes
dissolved in 10 mL distilled water and filtered
4.5-6.6 Moderately Warm water
polluted fishes into a 25 mL volumetric flask; 25 mL 18 M NaOH
Below 4.5 Highly polluted Cannot support solution with 5g KI and 0.15g NaN 3, 250 mL

0.125 M stock Na2S2O3, 500 mL 0.5 M H 2SO4.
Starch solution was freshly prepared by
weighing 1.0 g starch, dissolved, and poured in
approx. 100 mL distilled water and kept at 90°C-
100°C. The 250 mL 0.0125 M standard Na2S2O3
solution from 0.125 M stock Na2S2O3 was done
by group.
In the standardization of Na2S2O3, 0.15 g KIO3
was weighed using a 50 mL beaker and then
dissolved with distilled water. It was then
transferred quantitatively into a 100 mL
volumetric flask, diluted, covered with parafilm,
and mixed. 10 mL was taken out and put in
three Erlenmeyer flasks, labelled properly. 20
mL distilled water was poured in each flask. 1.0
g KI and 10 mL 0.5 H2SO4 was added in each
flask. Using Na2S2O3 as standard, with a
0.011298307 M, titrated the solution until pale
yellow. Immediately added 1.0 mL starch
solution and then continue titration until
complete disappearance of blue color.
Afterwards, standardization was performed in
triplicate.
In the analysis of the water sample, a syringe
was used to add 0.5 mL MnSO4 solution and 0.5
mL NaOH with KI and NaN3 solution without
producing air bubbles inside the sample then 2.0
mL of concentrated H3PO4 using a pipette was
added. The pipette should be below the surface
of the water. The solution was covered again
and mixed and left alone for 10 minutes. Aliquot
taken out was 50.0 mL and transferred in a 250
mL Erlenmeyer flask. Using Na2S2O3 as
standard, with a 0.011298307M, titrated the
solution until pale yellow. Immediately added
1.0 mL starch solution and then continue
titration until complete disappearance of blue
color. All sample triplicate were performed and
the collected data was expressed in 2.049 ppm.
Results and Discussions
The chemical reactions in this experiment are
−¿+6 H
+¿ → 3I 3+ 3H 2O +K
KIO 3−¿+8 I
(1)
−¿→S 4 O 6
(2) 2 S 2 O 32−¿+I 3
−¿→ Mn(OH )2¿
(3) M n2+¿+ 2( OH )
(4) 4Mn(OH)2 + O2 → 4MnO(OH) + 2 H2O
−¿→2 M n
6 H +¿+2 MnO (OH )+2 I
(5)
−¿¿
−¿ → I ¿
(6) I 2+ I 3
−¿ ¿
−¿→S 4 O6 + 3 I
2−¿+I 3
(7) 2 S 2 O3
The standard used for the titration was
thiosulfate (S2O3) and the pertinent chemical
R. J. C. De Guzman / Chemistry 26.1 (2017) Page |2
reactions are (1) and (2). In reaction (1), excess
KI was added to provide I which forms I 3.
−¿¿
Sulfuric acid (H2SO4) is only added after KI
crystals because doing the reverse would
jincrease the dissolved oxygen. Sulfuric acid
produces H+ ions and this is importance
because an acidic environment is needed for the
−¿ ¿
reaction with iodate ( IO3 ) to occur. The
whole process produces a dark yellow solution.
Standardization was continued in reaction (2)
when sodium thiosulfate was standardized
−¿¿
against I 3 . Titration was performed until pale
yellow color formed from the previous dark
yellow. Since it is difficult the determine the
endpoint through pale yellow color, 1° starch
solution was added which turns the solution to
deep blue. The rationale behind this is Iodine
gets adsorbed on the starch molecule surface
and product of adsorption has strong, blue color.
Starch was not added prior to titration because
starch solution undergo decomposition when
mixed with high I2 concentrations and this
results to a higher endpoint. (Titrations.info,
2014)
The solution is further titrated until deep blue
color turns colorless indicating that the
endpoint was reached. Table 1 shows the data
gathered from the standardization. The net
volume (mL) for all three trials was 37.0 and the
calculated molarity of the standard was 0.0113
M
Using the reactions (1) and (2), it can be
deduced that 1 millimole of IO3- is equivalent to
6 millimoles of S2O3.
Pertinent reactions for the analysis of the water
sample are (3) and (4) when water sample was
treated with MnSO4 and NaOH with KI and NaN 3
solution. Addition of MNSO4 solution can
−¿¿
precipitate with O H . Manganese alone
undergoes reduction and causes the volume of
+¿ ¿
¿ titrant used to decrease and thus DO content
¿
¿
will decrease. To counter this, KI precipitates
−¿ ¿
2−¿+ 3I ¿
M n2+¿¿ and produces I that will be oxidized
¿
¿ −¿¿
¿
to I 2. NaN3 on the other hand eliminates nitrites
+ ¿+ I +4 H O ¿
that converts I −¿¿to I 2 in the solution. In
2 2 2
¿
reaction (3), M n2+¿¿ will liberate O H
¿ −¿¿
from
added NaOH and thus forms Mn( OH )2
¿
¿
The sample was again sealed and then mixed
thoroughly and vigorously and formed the
precipitate 4MnO(OH) and this fixed oxygen
content in the sample.
 R. J. C. De Guzman / Chemistry 26.1 (2017) Page |3

In reaction (5), H3PO4 was added to provide

H +¿¿ ions generating an acidic medium which
dissolves 2MnO(OH) to 2 M n2+¿ ¿ and I 2 . The
sample was again mixed thoroughly and
vigorously to completely dissolve the oxygen
and accurately account for the DO content of the
sample. In reaction (6) I 2 reacts with excess
I −¿¿ and forms I 3.
Titration performed for the primary standard is
repeated this time for the water sample and this
References
is indicated by equation (7). Thiosulfate titrated
against I 3 will reduce it to I . Performed
−¿¿
Skoog, Douglas A., Donald M. West, and F. James.
calculations for the standard and the DO content Holler. Fundamentals of analytical chemistry.
can be seen in the appendix. 9th ed. Fort Worth: Saunders College Pub., 1996.
Print.
Conclusions and Recommendations
Petrucci, Ralph H. General chemistry: principles
The dissolved oxygen (DO) content of and modern applications. Toronto, Ont.: Pearson
the water sample is calculated to be at 2.049 Canada, 2011. Print.
ppm and it can be concluded based on the
results of the experiment that the body of water Soden, JM. "The Winkler Titration
where the sample was taken theoretically Method." Sciencing. Leaf Group, n.d. Web. 28
cannot support life because it is highly polluted Mar. 2017.
(See Table 1). Relative standard deviation is at
41.595 ppt and its confidence interval at 95% is Fondriest Environmental, Inc. “Measuring
2.048631741± 0.211547453 . Dissolved Oxygen.” Fundamentals of
Environmental Measurements. 7 Jan. 2014. Web.
These results are however deemed
anomalous since aquatic life do exist in the “Information about end point detection with
source of the water sample. It is hypothesized starch in iodometric titration.” Iodometric
that the DO content of the sample was titration - detection of end point witch starch, 22
significantly altered by heterotrophic organisms Mar. 2014, Web.
present in the sample. The sample was stored
for two days giving these organisms plenty of Clemente, Jamela Jirah L. “Redox Titration:
time to consume most of its DO content and thus Winkler Method for Dissolved Oxygen Content
leading to false results. Other than the Determination.” Diliman, Quezon City,
consumption of oxygen, the deviation of the DO University of the Philippines, University of the
content can be caused by errors such as altering Philippines, 2013.
the correct order of the addition of reagents,
inducing air bubbles during titration, or water Rivera, R.Y.A. “Quantitative Determination of
used in the experiment is not boiled. Dissolved Oxygen Content by Winkler Redox
Titration.” University of the Philippines,
To avoid such anomalous results in the University of the Philippines, 2015.
future, it is best that the water sample must be
subjected to Winkler titration immediately to
avoid addition or reduction of DO content. The
temperature of the sample should also be noted
to better compare accepted DO content for
water quality. Series of replications of the
experiment is also recommended to give a
precise data of the actual DO content in the
water source.
 R. J. C. De Guzman / Chemistry 26.1 (2017) Page |4

Appendix

A. Raw Data Table

Standardization of Na2S2O3 Solution

Primary Standard used: KIO3
Formula mass of 1 standard: 214 g/mol
%Purity of 1 standard: 99.4
Weight of standard, g: 0.15
Volume of standard solution: 150 mL

Trial 1 2 3

Volume of KIO3 Standard, mL 40 40 40

Final Volume Na2S2O3, mL 37.6 37.1 37.6

Initial volume Na2S2O3 0.6 0.1 0.6

Net volume of Na2S2O3 37.0

Average M Na2S2O3 0.0011298307 M

Sample Analysis
Location of sampling: Science Complex Park, Institute of Mathematics.
Total volume of sample: 400 mL

Trial 1 2 3

Final volume of Na2S2O3, mL 1.2 2.3 3.4

Initial volume of Na2S2O3, mL 0 1.2 2.3

Net volume of Na2S2O3, mL 1.2 1.1 1.1

DO Content, ppm O2 2.1693 1.9885 1.9885

Average DO Content, ppm O2 2.049

RSD 41.59451613 ppt

Confidence Interval 2.048631741± 0.211547453

B. Working Equations
 R. J. C. De Guzman / Chemistry 26.1 (2017) Page |5

0.15 g of KIO 3 6 mol Na2 S2 O3 0.010 Laliquot

[ Na2 S2 O3 ]std =
( 214.0
g
mol
KIO 3 )( 1 mol KIO3 )
( 0.994 ) (
0.1 Ltotal volume )
Volume of Na2 S 2 O3 , L

ppmO2=( L Na2 S2 O3used ) ( [ Na2 S 2 O3 ] std ) ¿ ¿

C. Sample Calculations

1. Molarity of the standard Na2S2O3 solution (All Trials)

0.15 g of KIO 3 6 mol Na2 S2 O3 0.010 Laliquot

[ Na2 S2 O3 ]std =
( 214.0
g
mol
KIO 3 )( 1 mol KIO3 )
( 0.994 ) (
0.1 Ltotal volume )
0.037 L Na2 S 2 O3

[ Na2 S2 O3 ]std =0.011298307 M 0.011 M

2. PPM O2 of the water samples (mg/L)

ppm O 2 ,trial 1=( 0 .001 2 )( 0.011298307 ) ¿ ¿

ppmO2 ,trial 1=( 0.001 1 )( 0.011298307 ) ¿ ¿

ppm O 2 ,trial 1=( 0 .001 1 )( 0.011298307 ) ¿¿

Average ppmO2=2.049 ppm

3. Relative Standard Deviation(RSD) and Confidence Interval

Standard Deviation 0.085211846

RSD= (1000 ppt )= ( 1000 ppt )=41.59451613 ppt
Mean 2.048631741

t ( standard deviation ) 4.30 ( 0.085211846 )

Confidence Limits ( 95 % confidence level )=mean ± =2.048631741 ± =
√n √3