Appendix D The Winkler Detennination1

H.L. Golterman 2

The most precise determination of dissolved oxygen can be carried out by a iodomet-
ric titration according to Winkler [15]. Although this method has been modified some-
what, the principle is unchanged and it is therefore probably the oldest method for
water analysis still employed.
The dissolved oxygen in the water combines with Mn(OH)2 forming higher hydro-
xides under alkaline conditions, which, on subsequent acidification in the presence of
r libarate 12 in an amount stoichiometrically equivalent to the original dissolved O2
content of the sample. There is doubt on the oxidation state of the oxidized manga-
nese hydroxide, but as eventually all electrons transported from the manganese to the
oxygen will be donated from the iodide to the manganese, this argument is not essen-
tial for the practical employment of the method. The reactions are as follows:

2 H2 0 + 4 Mn(OH)2 + O2 -+ 4 Mn(OH)3
or
2 H2 0 + 2 Mn(OH)2 + O2 -+ 2 Mn(OH)4
followed by

4 Mn(OH)3 or 2 Mn(OH)4 + (12 or 8) H+ + 41- -+ (4 or 2) Mn2+ +

+ (12 or 8) H2 0 + 2 12 .
The 12 is then determined by titration with (sodium)thiosulfate:

12 + 4 Na+ + 2 S2 O~- -+ 2 r + 4 Na+ + S4 O~- + 2 H+.

However, any other method of titrating the liberated 12 can be used equally well,
especially when potentiometric apparatus is available. (As 2 0 3 , e.g., has the advantage
of a stable stock solution and does not need to be standardized against another stan-
dard, as the thiosulfate; the pH has however to be between 4 and 9, which can be
easily adjusted with borax buffer solutions).
Normally, however, thiosulfate is used. The endpoint is often determined with the
blue color formed by 12 with starch.

1 This article combines relevant parts of Golterman et al. (1978) and Golterman and Wisselo (1981)
2 Station Biologique de la Tour du Valat, Camargue, Le Sambuc, F -13200 ArIes, France

Polarographic Oxygen Sensors (ed. by Gnaiger/Forstner)

©Springer-Verlag Berlin Heidelberg 1983
The Winkler Determination 347

Table 1. Concentrations of fixative and titrant solutions

Solutions Concentrations Add

Fixative mol dm- 3 a g dm- 3 per 100-150 cm 3

b 500 1 cm 3
MnS04 X 5 H2O 2
Alkaline iodide 10 400 g NaOH 1 cm 3
(with or without azide) 900 g NaI
(10-25 g NaN 3 )
Titrants
Na 2S2 0 3 X 6 H2O 0.0125 6.2 Depending on O 2 conc.
or and volume of (sub)sample
(with 1 pellet NaOH) 0.00250 taken
1/6 KI0 3 0.100 3.567

a mol in the new S.1. sense is used

b Most firms produce different hydrates; all can be used including MnCI 2. The concentration in
mol dm- 3 is decisive

In principle all compounds reacting with the 12 will interfere. One of the most like-
ly ones is nitrite. This interference can be eliminated by the use of NaN 3 (sodium
azide), up to concentrations of 0.36 mmol dm- 3 (= 5 mg dm- 3 N0 2-N). It must be
realized, however, that concentrations of 14 J1g dm- 3 of N0 2 -N, which is a rather high
value for unpolluted waters, is only equivalent to 0.1 % of the normally occurring oxy-
gen concentrations. This modification of the Winkler method is therefore only impor-
tant for partially purified sewage waters.
An important modification of the Winkler method was given by Pomeroy and
Kirschman [11], who advised a higher concentration of Nal than the reagent formerly
used. The advantages are that errors due to volatilization of 12 and to interference by
organic matter are reduced (because of the reaction 12 + 1- ~ I;), that the hydroxides
dissolve more readily, and that the starch end point is sharper. The possible interfer-
ence of organic matter being oxidized by the liberated 12 is often overlooked. Alster-
berg [1] proposed a bromination of the organic matter before the oxygen determina-
tion, while the excess Br2 is later combined with salicylate. This reagent must be pre-
pared freshly and must be colorless; it is best prepared from reagent pure, crystalline
sodium salicylate, which is commercially available. The Br2 destroys NO; as well, so
that NaN 3 is not required. The method is not often employed; its accuracy will be dis-
cussed below.
Greater precision than with the starch end point determination can be obtained
with potentiometric end-point detections, of which the best is the socalled dead stop
end-point technique (cf. Chap. III.l). In this method thiosulfate is added in an accu-
rately measured excess and is back-titrated with K10 3 ; an extra advantage is that the
12 occurs only for a very short moment rendering oxidation of organic matter less like-
ly. The reactions are as follows:

12 + 2 S20~- -)0 S40~- + 2 rand

103 + 2 Sz O~- -)0 r + S4 O~- and
 348 H.L. Goltennan

Fig. 1. Circuit diagram. (From Golter-

man et al. [5])

M1
R3
B1
I

The dead-stop end-point technique is an amperometric or potentiometric titration

with two identical Pt electrodes. One of these is made the indicator electrode by elec-
trical polarization with a small constant voltage, normally 50-100 mY; the other one
serves as a reference electrode. The electrodes are covered with very thin layers of H2
and O2 derived from the hydrolysis of water. Before the end point the current remains
constant (practically zero). As the thiosulfate-tetrathionate couple is not reverSibly
oxidizable, the electrodes cannot be depolarised and the thin adsorbed gas layers serve
as a barrier against the current. As soon as both 10; and 1- are present, oxidation and
reduction reactions become possible at the electrodes, the gas barrier (the electric re-
sistance) disappears, the adsorbed oxygen being reduced and the adsorbed hydrogen
being oxidized. Thus a sharp change in current and thus in voltage is found exactly at
the end point.
Dead stop titrators may be bought, but they can be made relatively easily. A circuit
diagram is given in Fig. 1. Relatively large Pt electrodes (gauze) are used here. Their
large surface enables the usage of low voltages and small currents, so that the apparatus
can easily be used in the field. The use of a precise micro burette is advisable. Best re-
sults will be obtained by graphing the current readings against volume of titrant as the
trace is always slightly curved at the end point. A linear extrapolation is possible using
points sufficiently distant from either side of the end point. With the apparatus of
Fig. 1 we found that this, however, hardly improved the accuracy.

1 Sampling

It is important to realize that the preciSion of the method depends as much on the
sampling procedure as on the chemical determination. For accurate work a displace-
The Winkler Detennination 349

ment sampler is necessary. The lower end of the outlet tube of the sampler is placed
above the bottom of the Winkler bottle. The contents of the sample bottle are dis-
placed about three times, before the sample is collected. The bottle must always be
completely filled and reagents or preservatives must be added immediately after sampl-
ing. If the samples cannot be titrated directly, they should be kept alkaline and be
stored in a cool place, but avoiding temperature changes. The samples must never be
kept after acidification; not even during the titration of other samples of a series.
For very shallow waters the sample may be taken in a syringe and analyzed by a
small-scale technique [4]. See further discussion in [5] and [10].
Recently Golterman and Wisselo [6] presented a ceriometric method for dissolved
oxygen and COD, following an older procedure using Ce 3+ [7). The principle is the
same as for the Winkler method. The advantages are that no volatile h is produced and
that the Ce4+ may be measured either titrimetrically or colorimetrically. Combination
with a COD estimation prevents all difficulties with organic matter. The chemical prin-
ciples behind this combined COD-0 2 technique are not so straightforward as the
Winkler method, the method is therefore not so easily understandable in the beginning.

2 Precision and Accuracy

The accuracy depends very much on the sampling precision and on the accuracy of the
concentration of the titrant, i.e., the thiosulfate. Normally the concentration of the
thiosulfate decreases slightly; this can be diminished by the addition of 0.5 g dm- 3
of Na2C03. The accuracy is high, as the method is based on stoichiometric reactions
[3] and not on a calibration curve, while other possible factors are insignificantly
small.
There are several papers about the preciSion of the Winkler determination; seldom,
however, is the precision of this determination discussed for samples of natural waters
(Chap. III.1).
Rebsdorf [12] found that the Alsterberg modification gave the same results as the
Winkler method in unpolluted waters and higher results - but still too low - in pollut-
ed waters, which he attributed to oxidation of organic matter in the normal Winkler
technique. He found the Alsterberg technique to have a lower precision than the Wink-
ler.
A comprehensive review of modifications of the Winkler method [8] pointed out
that several modifications lead to no improvement because they introduce new sources
of error, decreasing the precision.
Bryan et al. [2] used a photometric titration by measuring the changing absorption
of UV radiation. They mention a coefficient of variation of 0.1 % for seawater; it
seems likely that this will be less good in (eutrophic) freshwater.
Mackereth et al. [9] found a relative standard deviation of 0.6% (n = 7) for the nor-
mal Winkler titration for standard solutions. Talling [13] used an amperometric end-
point detection and found a reproducibility (mean minus extreme) for iodine solutions
equivalent to 0.6 J,Lmol O2 dm- 3 .
350 H.L. Golterman

Carpenter [3] found an accuracy of 0.1 % for the Winkler method, proving a stoichi-
ometric reaction. He discussed precision of different iodometric end-point determina-
tions, but not the precision of the Winkler method.
Tschumi et al. [14] mention that the precision of the Winkler method is often quot-
ed as 0.6 JlIllol dm- 3 and believed that this value refers to the standard deviation. The
standard deviations in their study (using a photometric end-point detection) were
0.06 JlIllol dm- 3 (0.02%) for H2 0, 0.5 JlIllol dm- 3 (0.15%) for lake water containing
0.76 mm 3 phytoplankton per dm 3 and 1.0 ~mol dm- 3 for lake water containing
7.07 mm 3 dm-3 . The influence of the phytoplankton density on the precision is strik-
ing: 1.6 JlIllol dm-3 as largest difference for diluted lake water and 3.4 ~mol dm- 3 for
their concentrated lake water. We have found the same phenomenon for the classical
Winkler determination and think that this precision is controlled by the adsorbance of
12 on the phytoplankton, which, although reversible, may not be sufficiently rapid for
high precision. Therefore I feel that much more attention should be given in the dis-
cussions on the precision of this method to the phytoplankton density and that gen-
eralizations may be dangerous. The 12 adsorption influence is overcome by an electro-
chemical end-point detection of a back titration of an accurately known excess of
thiosulfate, which thus presents optimal conditions. Results that are possible with this
method in lake water containing a high phytoplankton density are given in Table 2.
From this table it can be seen that in 11 out of 15 cases the mean value differs by 0.6
to 0.9 JlIllol dm- 3 from the extreme value. If a manual dead stop method is used (Fig.
1) this difference is about 0.6 JlIllol dm-3 . The pooled standard deviation of the series

Table 2. Results of Winkler determinations chosen at random from routine measurements. The
Winkler determinations were carried out with an automated dead stop titration following proce-
dure 8.1.4 of the I.B.P. manual Nr 8 (Golterman et al. [5])
All samples were from Tjeukemeer, and had chlorophyll a concentrations of about 100 mg m-'

Results in mg dm-' of O2

9.66 11.25 11.68 9.70 11.38 11.60 11.71 10.78

9.63 11.30 11.68 9.70 11.38 11.78 11.78 10.82
9.73 11.27 11.63 9.68 11.44 11.77 11.68 10.81
0.06 0.027 0.033 0.013 0.04 0.12 0.057 0.23 mean - extreme

10.88 11.59 11.26 11.60 12.90 13.34 10.12

10.91 11.53 11.26 11.65 12.91 13.36 10.11
10.91 11.60 11.32 11.63 12.93 13.35 10.13
0.02 0.043 0.04 0.027 0.017 0.017 0.01 mean - extreme

From these values it can be seen that in 11 out of 15 cases the mean value differs by 0.02-
0.03 mg dm- 3 from the extreme value. If a manual dead stop method is used the difference is about
0.02 mg dm- 3 • The pooled standard deviation of this series is 0.04 mg dm- 3 or 0.025 after exclusion
of columns with s > 0.05. The standard deviation was also measured directly on 12 samples. The
following values have been found:
11.75 11.73 11.74 11.80 11.74 11.78
iii = 11.77
11.72 11.72 11.83 11.77 11.81 11.80
In this series s = 0.038 and the coefficient of variation 0.3%
The Winkler Determination 351

of Table 2 is 1.3 ~ol dm- 3 or 0.8 after exclusion of columns with s > 1.5. The stan-
dard deviation was also measured directly on 12 samples, in which the standard devia-
tion s = 1.2 and the coefficient of variation = 0.3% (mean value was 367.8 ~ol dm- 3 ).
Therefore I do not believe that the quoted 0.6 Slmol dm- 3 is the standard deviation,
but that it is the likely difference between lowest (or highest) value and the mean
value in a triplicate series under good, but not optimal conditions. It seems further-
more that Tschumi et al.'s precision is higher than the normal Winkler technique, but
not higher than the most precise versions. For the normal technique a precision of
0.5%~.6% seems to be likely, but may be influenced by phytoplankton density.
With a titrimetric method we found no difference in standard deviation of tap-
water and of lake water with little or much phytoplankton.
The standard deviation for the proposed Cerium technique [6] was about 1%, but
the method was designed to be rapid and to be used in organically rich waters.

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30-75
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