Water Research Vol. 15, pp. 449 to 456, 1981 0043-1354/81/040449-08102.

00/0
Printed in Great Britain Pergamon Press Ltd

DETERMINATION OF OZONE IN WATER BY

THE INDIGO METHOD
H. BADER and J. HOIGNI~
Federal Institute of Water Resources and Water Pollution Control (EAWAG), Swiss Federal Institute
of Technology, CH-8600 Diibendorf, Switzerland

(Received 20 May 1980)

Al~tract--The concentration of aqueous ozone can best be determined by the decolorization of indigo
trisulfonate (600 nm, pH below 4) whenever the ozone cannot be measured directly by its u.v. absorp-
tion. The method is stoichiometric and extremely fast. The change of absorbance vs ozone added is
-2.0 _ 0.1 x 104 M -1 cm -~ and is independent of the concentration of aqueous ozone in the range
0.005-30 mg 1- t. The precision of the analysis is 2% or 3/~g 1-1 for low concentrations if a spectropho-
tometer or a good filter instrument is used. Visual methods can be used to measure 0.01 mg 1-1 ozone.
Secondary oxidants produced by ozone in natural water, including hydrogen peroxide or chlorite, do
not interfere; chlorine can be masked. The reagent solution is stable for 3 months. The method is
recommended for kinetic measurements, for studies of ozonation processes and for visual field methods.

1. INTRODUCTION merciaUy available reagents, listed as pH or redox

Approximately a dozen analytical methods for the de-
termination of aqueous ozone have been described in
the literature and about half of them are still applied.
However, most of these methods are not specific and
often give incorrect results.
Among the first reagents used to measure ozone in
air or exhaust gases were indigo and its water soluble
derivatives, the sulfonated indigo compounds such as
indigo disulfonate, indigocarmine and indigo trisulfo-
hate (Thenard & Thenard, 1872; Fonrobert, 1916;
Dorta-Sehaeppi & Treadwell, 1949; Dr~iger &
Driiger, 1966; Verein Deutscher Ingenieure, 1977). In
contrast, the application of the indigo methodfor the
analysis of ozone in drinking water has been neg-
lected. We can cite 3 reasons for this: (1) In most
waterworks ozone is measured by merely adapting
analytical methods already accepted for determi-
nation of chlorine or other oxidants; (2) Most analyti-
cal chemists are sceptical about methods based on
decolorizations; and (3) It was not recognized that
erraneous results are often only due to a preliminary
decomposition of ozone and that this can easily be
avoided by maintaining pH values below 4 (Hoign6 &
Bader, 1976).
In the past few years we have accumulated experi-
ence on the application of the indigo method for ana-
lyzing aqueous ozone in many different types of
waters (Hoign6 & Bader, 1979a, b, 1980). For these
studies this method showed many advantages: it is
very sensitive, precise, fast, specific, and easy to
handle. Since other analytical methods have presented
difficulties, many research and process chemists have
approached us for more detailed information con-
cerning this method.
Indigo is a well known classical blue vat dye
(example: Blue Jeans). Its water-soluble derivatives,
indigo disulfonate and indigo trisulfonate, are corn-
449
indicators. Aqueous solutions absorb light at about
600 n m with a rather high molar absorptivity (Preisler
et al., 1959, reports 2max = 605 rim, ~ = 23,800 M - 1
cm-1 for indigo trisulfonate).
The indigo molecule contains only one C----C
double bond which can be expected to react with
ozone with a very high reaction-rate constant (see
Fig. 1). At low pH the amino groups are protonated
and therefore unreactive (Hoign6 & Bader, 1978,
1979a). Thus, it can be assumed that one molecule of
sulfonated indigo reacts with one molecule of ozone
under proper reaction conditions including high rate
of mixing. Ozonolysis of the reactive ~ bond pro-
duces sulfonated isatin and similar products and elim-
inates the absorbance at 600 nm as shown in Fig. 2
(Dorta-Schaeppi & Treadwell, 1949).
2. EXPERIMENTAL
2.1 Chemicals
Indigo recu3entfor determination of ozone. Generally, the
stock solutions of Indigo Reagent were prepared by dis-
solving 0.6 g l- ~ (1 mM) potassium indigo trisulfonate in
20mM phosphoric acid. These solutions were replaced
when their absorbance at 600 nm had decreased to below
80% of the initial value (typically after 3-4 months storage
in transparent glass bottles). Potassium indigo trisulfonate
was generally purchased from Riedel-de Hahn AG (Redox
Indicator), but a product from Merck, which had been
stored for 10-20 years, gave similar results. The micro-
analysis for C/N/O and titrations with permanganate indi-
cated an indigo content of about 80% of the theoretical
value. (An uncertainty of the concentration of the Indigo
Reagent does not influence the accuracy of the analytical
method which is based on standard curves for changes of
absorbance achieved per added mole of ozone.)
o-Tolidine hydrochloride and DPD (N,N-diethyl-l,4
phenylene-diamine sulfate (GR) were Merck p.A. products.
(Comparable results were also achieved with the DPD test
kit "AQUAMERCK Nr. 11100 for chlorine".) ACVK (Acid
chrome violet K, Michrome Nr. 1047) was an Edward
450
so~
0 H~Nr--~
s o , . I", . . , ~IIl , ~ =| L "'1~j~' ' ~ ~" s o ,
o [, o.e2oooo .-' cm-' 1
1 ~'03
0 0.5-5 cm path length were used. A VITATRON photo-
sc~../,~.~k..~.O
2x II ".T r and corres~ndimg products
"¢'----"'"
(so~)
etc
Fig. 1. Indigo trisulfonate and its ozonation product.
Ascff
0.~
O~
0,~
02
i-
[]11 ,,
0.0
4O0 500
Fig. 2. Absorption spectra of Indigo Reagent after stepwise
ozonation (Indigo trisulfonate added: 10/tM).
Gurr Ltd (London) product; Phosphate Buffer Reagent for
pH 2: 28g N a H 2 P O 4 ' H 2 0 and 35g H s P O , (85V.) dis-
solved in t 1. dist. water. Other reagents were of commer-
cial pro analysis grade.
Aqueous ozone. Stock solutions of aqueous ozone were
prepared by continuously bubbling ozonated oxygen
(about 4%) through a gas-washing bottle into distilled
water chilled to 2°C. This stock solution generally con-
tained a steady-state concentration of ozone of 40 mg 1- ~.
Lower concentrations were achieved by dilution with dis-
tilled water. Whenever not required for later use, this water
was acidified with phosphoric acid to pH 2. When the
bottle was kept closed the concentration of ozone de-
creased less than 0.2~ per min (Hoign6 & Bader, 1978).
The aqueous ozone was transferred into the individual test
bottles with glass pipettes or by plastic dispensers. The
concentration of the diluted ozone water was determined
before and after the samples were withdrawn. These cali-
brations of the stock solution were performed by measur-
ing the u.v. absorbance of ozone at 258 rim. They were
based on a molar absorptivity of 2900 M - * c m - * which
was determined in other studies by a direct comparison
with the iodine method, here considered as the primary
standard (Kilpatrick & Herrick, 1965; Hoign6 & Bader,
1976). For these calibrations t h e samples of aqueous ozone
was acidified by sulfuric acid to give 0.3-0.5 M acid. About
0.2 g o f potassium iodide were added. The liberated iodine
was titrated with 0.1 M sodium thiosulfate using starch
indicator.
Care was taken that the aqueous ozone was always
handled in exactly the same way, whether it was trans-
ferred into samples containing tndigo g ~ t or inlo the
u.v. cells or into flasks for the iodide t i t r a t i ~ , All concen-
trations of ozone are therefore finally based on the iodide
H. BADER and J. HOIGNI~
method; the u.v. method was applied only as a more con-
venient secondary standard.
indigo trisuffomc acid
2.2 Instrumentation
Generally, the absorption measurements for the indigo
method were performed at 600 nm on a Beckman DK 2A
spectrophotometer which was equipped with a thermo-
stated cell compartment maintained at 25°C. Cells of
isatin sutfonic acid meter equipped with a 4 cm cell and a 595 nm interference
filter of 15 nm bandwidth was also used. In experiments to
I~.,-0.0 M-' cm-' I test the indigo method as a visual field method, pairs of
graduated glass cylinders were used (volume 250 ml. height
25 cm).
2.3 Glassware
Best reproducibilities were achieved when the glassware
n.Ri -I ozone o~ded used for handling of aqueous ozone was conditioned by
repetitive use for the same procedures.
2.4 Experimental procedures for the indigo method used with
a spectrophotometer
During this study, many varied procedures were tested.
Only illustrative examples can be given here:
Example for the determination of aqueous ozone in the
concentration range 0.3-15 mg 1- t : Series of 100 ml volu-
metric flasks were prepared. To each flask of the series
10 ml of 0.5 M Phosphate Buffer Reagent for pH 2, 1 ml of
1 mM Indigo Reagent and, in case of the measurements of
high concentrations of ozone, about 20 ml of distilled water
were added. Some of these flasks were used to determine
. the standard curve. For this, individual flasks of the series
600 700 nm were dosed with different amounts of diluted stock sol-
utions of aqueous ozone containing about 2 m g l - ' ozone
e.g. 0, 5, 10... 25 mt, at random to minimize systematic
time drifts of ozone concentrations. This solution was
dosed with a glass pipette whose tip was placed below the
surface of the reagent solution while stirring with a Teflon
covered magnetic bar. The concentration of the ozone in
the diluted stock solution applied was calibrated simul-
taneously by measuring its u.v. absorbance at 258 nm in a
5 cm u.v. cell. Finally, distilled water was added to the
100 ml mark. During this operation the stirrer was tempor-
arily raised above the mark. The absorbance of the residual
indigo present in these series of test solutions was
measured at 600 nm in a 5 cm cell. The values were plotted
vs the amount of the ozone added. A typical standard
curve is presented in Fig. 3(B). The determination of the
ozone in the unknown samples is thereafter performed
similarly by dosing specific volumes of these samples to
others of the prepared flasks of the series. In order to be
within the range of the amount of Indigo Reagent used the
size of the sample was selected to contain an estimated
10--40#g ozone. For example, when the concentration of
ozone was expected to be 0.5 mg 1-1 20-80 ml were
injected and when the concentration was very high, such as
15 mg 1-t. 1-2.5 ml were dosed. The volumetric flasks were
filled again with distilled water to the I00 ml mark. The
residual absorption at 600 nm was compared with the stan-
dard curve and the original concentration of ozone was
calculated.
Comparable procedures were adapted to determine
ozone in other concentration ranges. In cases of higher
concentrations, higher amounts of the Indigo Reagent were
dosed. In this case the determination of the residual
absorbance of the Indigo Reagent was determined using
0.5 cm cells. An example of a standard curve is shown in
Fig. 3(A). In this case the amounts of the Indigo Reagent
and of the ozone applied were about 10 times higher than
those in the example shown in Fig. 2. When lower concen-
trations of ozone were determined, less Indigo Reagent was
used. Figure 3(C) gives an example in which the concen-
trations were 4 times lower than those for the example in
 Determination of ozone in water by the indigo method 451

5iS
A4©m

0.04
08' ~, (A)
0.03

0.02
06 •~
"\ 001
0A ~ I I t I~'lk ! IL
(200 5 IO )5 20 25
Added [ 031/zg I-)
0.2
Fig. 4. Standard curve absorbance vs ozone added to
Indigo Reagent. Measurement with filterphotometer. Inter-
O0 I I i ,.I
I 2 3 4~ ference filter 595 nm. Cone. of Indigo Reagent 0.6 M. (Con-
Added[03] n ~ f' centrations are based on final volume.)

o8~.\
- "\.\ (B)
glass cylinders A and B. Cylinder A was filled to its 25 cm
mark with the sample of aqueous ozone. Cylinder B was
filled with distilled water. Cylinder B was then emptied
into an Erlenmeyer flask and refilled with this reference
solution until the color intensity, when observed from top,
matched that of cylinder A. A standard curve was obtained
04 •~e by repeating such measurements using different known
amounts of ozone. The height of the reference solution in
cylinder B was plotted vs the concentration of the aqueous
02 ozone applied in cylinder A. A standard curve is given in
Fig. 5. Thereafter, the concentration of ozone in the
samples was determined by comparing the height of sol-
(20 I
o.i
I
0.2
I
0.3
"N ~
0'~ ution in the reference cylinder with the heights given in the
Added [ 03] mg I standard curve. The size of the sample was taken into
IlOOem account for the calculation of the concentration.
At=,, I
o.o,. ~ ICl 2.6 Experimental procedures applied for other spectrometric
analysts of ozone
For some tests, ozone had to be determined by the direct
0.03 measurement of its u.v. absorption at 258 nm even in
natural waters. In these waters, which sometimes exhibited
significant background absorption at 258 nm, the measured
absorbance was corrected by determining the background
002 absorbance after immediately destroying the residual
ozone with an approx. 10-fold excess of aqueous nitrite or
butene-ol.
The ACYK method was carried out as described by
oo
Masschelein & Fransolet (1977). The methods based on

I I • i~ h• Approx., A 2~~,.
6 0 Ohm
O. 02 0.04 0,06 (208 O I- ='4
Added [031 mg I-' i
25 • -~o. eo
Fig. 3. Standard curves o f absorbance (A) at 600 nm vs
ozone added to Indigo Reagent. Measurement with 2o .!o.
s t ~ r o p h o t o m e t e r . Conc. of Indigo Reagent added: (A):
0.1 raM; (B) 10 #M; (C) 2.5/~M. (Cone. of Indigo Reagent
and ozone are based on final volumes.)
15~ ~0.48

,e ax -~o32
,

Fig. 3(B). Figure 4 gives an example for a 14 times lower S ,~ ~ ,,o.)6

concentration range which has however measured on a %** I
filter photometer. 0 I I I I i I 1% i
If the rates of fast consumptions of ozone were measured 20 40 60
(Hoign6 & Bad•r, 1981), the Indigo Reagent was injected Added [0~]/~0 I-I
directly into the flasks containing the aqueous ozone. Ozo-
nation reactions were thus immediately stopped upon mix- Fig. 5. Standard curve for visual method. To both
ing with the Indigo Reagent. The concentration of the cylinders of 250 ml 0.4/zmol of indigo trisulfonate had
ozone was determined again from the decolorization of the been added, h* Is the height of reference solution in
Indigo Reagent as described above. cylinder B giving the same visual color intensity as 25 cm
of the solution in cylinder A which was decolorized by the
2.5 Visual method addition of a sample of aqueous ozone. The right-hand
A visual method was developed for field studies: The scale indicates the approximate absorbance of the solution
same amount of the Indigo Reagent was added to a pair of in cylinder A. (Cone. of ozone is based on final volume.)

w.R. 15/4--D
452 H. BADERand J. HO1GNE
o-tolidine and DPD were performed as described in Stan-
dard Methodsfor the Examination of Water and Wastewater
I1975).
3. RESULTS
3.1 Absorption spectra of the Indigo Reagent
The Indigo Reagent shows an absorption maxi-
mum at 600 nm. The apparent absorption coefficient,
based on the amount of total mass of commercial
(non-purified) reagent added is 16,500 M - 1 cm- 1. The
shape of the absorption curve and )~,x do not change
with ozone addition (see Fig. 2).
3.2 Sensitivity and precision of the indigo method as
used with spectrophotometers
The absorption of samples containing the Indigo
Reagent decreased linearly with the amount of ozone
added over all the concentration ranges investigated.
From the slopes of standard curves the photometric
sensitivity of the analytical method may be deduced.
Values are presented in Table 1. The absorbance
change at 600nm is 2.0 + 0.! x 104cm -1 per mol 1-1
of added ozone for all concentrations tested. Lack of
precision may be due to uncertainties in the calibra-
tion of aqueous ozone applied for the determination
of the standard curves. We estimate that this uncer-
tainty can cover a range of 5% when series performed
at different concentration ranges are considered. The
precision of the slope of single standard curves (about
8 points measured) was generally within this range.
Neither deterioration of the absorbance of the Indigo
Reagent solution to 80% of its original value during
extended storage (3 months) nor a partial decoloriza-
tion by a preliminary ozonation affected these calibra-
tions.
The error of the ozone determination decreases
with the relative amount of decolorization and
increases in case of very low concentration ranges.
However, the relative standard deviations calculated
from extended series were generally about + 2% when
the concentration of ozone was above 0.1 mg 1-1 and
when the residual absorbance was in the region of
10-60% of the initial absorbance. The main part of
Table 1. Typical sensitivity values of the Indigo Method
Range of ozone
co dose AA*
(/~M) (mg 1-1) (M - l c m - l)
100 0.2-3.5 (2.02 _ 0.10)' 104
10 0.02-0.3 (2.02 _ 0.10)" 104
2.5 0.02q?.l (1.99 +_0.10)" 104
Co = cone. of potassium indigo trisulfonate applied,
based on final volume.
AA = C h a n g e o f a b s o r b a n c e ( c m - 1) at 600 n m m o l - 1
1- t added o z o n e .
* Estimated range of accuracy of calibrations =
+ 0 . 1 . 1 0 ' * M - l cm -1.
the variances is thereby expected to be due to the
variance in ozone doses.
The effect of the rate of mixing of the aqueous
ozone with the Indigo Reagent solution was tested in
special series. Only a small difference of the sensitivity
factor (2°; between matched pairs of extended series
of simultaneously prepared samples) could be found
between normally and extremely slowly stirred
samples.
3.3 Measurements with filter photometers
Measurements of the residual absorbance of the
Indigo Reagent with a photometer equipped with a
595 nm filter gave results and variances comparable
to those determined with the spectrophotometer. An
example of a standard curve is given in Fig. 4. The
slope of this curve is 85°,0 of that expected from
measurements using a spectrophotometer at 600nm,
probably because the 595 nm filter does not quite
match the maximum of the absorption spectrum and
the filter band-width of about 15nm exceeds the
maximum of the absorption band of indigo trisulfo-
nate (c.f. Fig. 2).
3.4 Visual methods
An example of results found by the visual method is
presented in Fig. 5. An incremental ozone dose of
10 pg 1- 1 can be easily detected when 25 cm cylinders
are used.
3.5 Stoichiometricfactor
Extrapolation of the standard curves to the inter-
cept on the absissa shows that only 80% of the com-
mercial indigo product applied is effective in consum-
ing ozone. This calculation is based on the assump-
tions that the potassium indigo trisulfonate applied
was 100`0/0 pure and that 1 mol ozone added would
decolorize 1 tool of indigo. Above pH 4 the stoichio-
metric factor often decreases and it seems to become
somewhat erratic.
3.6 Rates of reactions
For the application of the Indigo Method for kin-
etic measurements it is important to note that all the
ozone is consumed by the reagent immediately upon
mixing. This was observed on a 1 s time scale even in
systems where the concentration of ozone decreases
by fast concurrent reactions, e.g. 20% s-1, and even
when the concentrations of both the indigo trisulfo-
nate and the ozone were, at the time of mixing, as low
a s 10 - 7 M {Hoign6 & Bader, 1981). That means that
the rate constant for the reaction of ozone with indigo
trisulfonate must be higher than 107M - is-1.
Aqueous indigo trisulfonate which is preozonized
until just decolorized consumes further ozone only
slowly. The apparent reaction-rate constant of t h e s e
ozonolytic products is about 2500 M - ts- 1 (Hoign6 &
Bader, 1981). Based on this, we may assume that t h e
indigo method will not be significantly disturbed by
short temporary local overdoses of ozone.
 Determination of ozone in water by the indigo method 453

03oddN
~nm
-- As.~ O25(mg I"~)
~7t
O.
-- aCVK A~" o~
DPD - - - o-Tolidine A=~'~'~ O~15
o61- .y ....... om A~'L" oz~
/
I_ // IACVK
O.5b.. . . . . -~ \ .............
]"'x,,x~" . - ' " Indigo
0.41'- "" k."~,~

0.3
0.2 I-
t ""
" " --..¢. o-Tolidine
~ "~ ~ .~......
N

O.f I I ~1" ~__

J 2
Time of reoction~h
Fig. 6. Stability of residual absorbance of different reagents after addition of aqueous ozone. In case of
ACVK and Indigo the amount of ozone added decolorized about 40% of the added reagent.

3.7 Temperature effect on absorbance mgl-,

The absorbance of the Indigo Reagent and the sen- 03

ffteOS.
sitivity factor for the determination of ozone de-
creases 2.0% with an increase of 10°C. O.fl

3.8 Stability of residual absorption after ozone addition 0.6

j a
The residual absorbance of Indigo Reagent 0.4
measured after an addition of aqueous ozone in dis-
0.,~
tilled water does not change during a 6 h time period.
A similar stability was only found with ACVK. In 0£ I I I
20 40 60
contrast, D P D and o-tolidine show significant ab- Time offer indigo odd. ~h
sorbance changes even when observed within I h
(ef. Fig. 6). a Rowwoter,loke Zurich (3Ore) I rng fJDOC
Row woter, G r e i f ~ , surfoce ~4rag F'DOC
3.9 Determination of ozone in natural water
Comparable results for the determination of the Fig. 7. Stability of residual absorbance of Indigo Reagent
residual ozone present in ozonized natural water have in different types of raw waters, expressed in apparent con-
centrations of ozone determined. The residual absorbance
always been found by measuring its absorbance at was about 50% of the non-ozonized reference.
258 nm (reference method) and by the indigo method.
Comparisons are presented in Table 2. The direct u.v.
measurement, however, required a rather complicated due rather to errors involved in the data of the refer-
procedure to compensate for the background absorp- ence method applied.
tion of natural water at 258 n m (see Experimental), In some types of ozonated surface water the resi-
also its sensitivity is relatively low (see Discussion). dual indigo trisulfonate shows a slow post-decoloriza-
Discrepancies between the two methods are therefore tion when observed over several hours. Samples of

Table 2. Comparison of cone. of residual ozone determined in different types of

ozonized waters by the direct method (u.v. 258 nm) and by the indigo method
(600 nm)
U.V.2sanm Indigo6oonm
Water (rag 1-1 ozone) (mg 1- l ozone)*

Bidistilled 0.51 0.53

Lake Zurich
(lake water, DOC 1.1 mg 1-1) 0.61 0.60
(after sandfilter, DOC 1.2 mg 1-~)1" 0.35 0.34
0.80 0.83
1.24 1.28
Greifensee (lake water, DOC 4 mg 1-1) 0.43 0.43

* All calibrations are based on a sensitivity factor of 20,000 M - ~ cm- 1.

t Ozonizations performed in plant at Zurich-Lengg.
454 H. BADERand J. HOIGNt~
water from Greifensee--a highly eutrophic lake
exhibited the maximum rate of change encountered in
our survey studies (Fig. 7). Such post-decolorizations
are, however, very small when purified drinking
waters are considered.
3,10 Effect of other oxidants
At pH 2 hydrogen peroxide, chlorite, chlorate and
perchlorate do not decolorize Indigo Reagent when
observed within a few hours and when the concen-
trations considered are within a factor of 10 of that of
the ozone to be determined. However, chlorine and
iodine lead to a slow post-decolorization. Their effects
increase with time. Chlorine can be successfully
masked by adding 500 mg 1-1 malonic acid to the
Indigo Reagent; malonic acid at pH 2 consumes
ozone with a reaction-rate constant of only 3 M ~s-
(Hoign6 & Bader, 1981) and therefore does not com-
pete with the Indigo Reagent for reaction with ozone
when added simultaneously with the reagent. An
example of the effect of chlorine in the absence and
presence of malonic acid is given in Table 3. Products
of ozonolysis of organic solutes seem generally not to
interfere with the residual Indigo Reagent when tested
within a few hours: no decolorization was observed in
any of the solutions we used for kinetic measure-
ments. This study covered many different types of
aromatic hydrocarbons, phenols and other products
(Hoign6 & Bader, 1981). However if, for example,
l mM 1-hexene-4-ol has been preozonized with
0.5 mM 03 at pH 2, it decolorized a 0.01 mM sol-
ution of Indigo Reagent significantly within 40 min.
Permanganate decolorized Indigo Reagent in a fast
reaction.
3.11 Indigo Reagent based on indigo disulfonate
All results reported here are based on an Indigo
Reagent which was prepared from commercial potas-
sium indigo trisulfonate. In response to inquiries
received, some tests have been repeated with an
Indigo Reagent based on commercial types of sodium
indigo disulfonate. Comparable results have been
Table 3. Effect of chlorine on the determi-
nation of ozone. The residual absorption (A)
was measured 4 rain and again I h after the
ozone (and chlorine) was dosed to the Indigo
Reagent. Malonic acid (m.a.+) was added di-
rectly to the Indigo Reagent before reaction
600 n m
0 3 CI2 m.a. Ascm
0.2 mg1-1 l m g l -~ 500mg1-1 4min lh
- - - 0.88 0.88
+ - - 0.51 0.52
+ - + 0.53 0.52
+ + - 0.44 0.24
+ + + 0.48 0.48
- not added.
+ added.
found for this reagent when the absorbancc of the
disulfonate was measured at 610nm, where this re-
agent shows its maximum absorption (Hoign6 &
Bader, 19801.
4. DISCUSSION
4.1 Stoichiometric Jactor for the decolorization of
Indigo Reagent
The sensitivity of the decolorization of Indigo
Reagent stays constant over a wide range of ozone
doses. This can only be expected if it is assumed that
the sulfonated indigo is a spectroscopically uniform
material with respect to its decolorization reaction; in
the presence of reactive impurities (such as other
indigo derivatives), both the absorption maximum
and the sensitivity could shift during ozonation.
An apparent stoichiometric factor of 1:0.8 for the
decolorisation of the Indigo Reagent agrees with the
microanalytical characterization of the commercial
potassium indigo trisulfonate. The phenomenon of
the stoichiometric factor decreasing above pH 4 can
be explained: in the presence of organic materials
with structures similar to that of the ozonolytic
products of indigo, a catalytic chain reaction leads to
an accelerated decomposition of ozone (Hoign6 &
Bader, 1976). Such chain reactions are initiated by
hydroxide ions and therefore proceed at elevated pH
during mixing of the reagents wherever temporary
local excess of ozone is present in the reaction mix-
ture. The secondary oxidants formed from decom-
posed ozone, OH radicals, do not however selectively
oxidize the chromophoric structure of residual indigo
(Hoign6 & Bader, 1978).
4.2 The sensitivity and limits of detection
The sensitivity, which amounts to a change of the
absorbance of AA = - 20,000 cm- 1 mol- t ozone
added l-~ is quite high. If a classical type of filter
photometer or u.v. spectrograph is used, a limit of
detection of about 5 pg I-1 can be achieved. When
working with a high performance spectrophotometer,
a detection limit of 1 #g l- ~ is approached (cf. Hoign6
& Bader, 1981a). Here the handling of the ozonated
water and not the instrumental accuracy become
limiting.
The detection limit of the simple visual method is
10 pg l- 1 when 25-cm cylinders are used. This limit of
detection fulfils the requirements for field studies on
drinking water: e.g. according to a recommendation
accepted in Switzerland, less than 50/~g1-t of resi-
dual ozone should appear at the consumers tap
(Schweiz. Lebensmittelbuch, 1972).
4.3 Sensitivity of the Indigo Method compared with
other spectrophotometric methods
In Table 4, the absorption regions and sensitivities
of different photometric methods are listed for corn-
 Determination of ozone in water by the indigo method
Table 4. Sensitivities of different colorimetric methods
for the determination of aqueous ozone expressed as
AA (change of absorbance per 1 mol 1-t ozone added
per I cm)
Wavelength AA
Method (nm) (M- 1 cm- t) Ref.
u.v. of 03 258 +2900 a
Indigo 600 - 20,000 b 4.4 Dynamic range of the Indigo Method
ACVK 550 - 2500 c
Orthotolidine 440 + 32,000 d The ozone dose must be adjusted to decolorize
DPD 510 + 15,000 e 20-903/o of the Indigo Reagent. For one fixed concen-
Iodide/triiodide 352 +25,000 f tration of the reagent and one predetermined relative
a--Kilpatrick (1965) and Hoign6 & Bader (1976).
b---this study.
c--own measurement; method of Masschelein (1977).
d--own measurement and Sulzer (1958).
e--own measurement, method of Palin (1975).
f---talc, from data of Shechter (1973).
parison. The most straightforward method is the di-
rect measurement of the absorbance of aqueous ozone
at 258 nm. This absorption occurs, however, in a
region where many types of other solutes interfere: in
natural waters, e.g. humic materials often cause a sig-
nificant background absorption which varies with
ozonation. In addition, the instrumental measurement
must be performed at the time at which the concen-
tration of ozone is of interest. Moreover, due to the
low molar absorptivity of ozone, the sensitivity of this
method is relatively low. This direct method, however,
must still be recommended as the best defined and
useful reference method for the calibration of ozone
concentrations whenever the limitations of sensitivity
and background absorptions are not relevant.
In contrast, the Indigo Reagent absorbs light in a
spectral region where only a few other substances
interfere. Natural waters, for example, have generally
no background absorption in the 600 nm region. The
sensitivity, when expressed as the change in absorb-
ance per added ozone, is quite high. In addition,
instrumental sensitivities are generally high at 600 nm
and even the sensitivity of the human eye is relatively
high at this wavelength. For most types of waters
measurements can be performed even hours after the
ozonated water has been added to the reagent sol-
ution.
The sensitivity of the indigo method is about 8
times higher than that of the ACVK method. In ad-
dition, measurements at 600nm are preferable to
measurements at 550 nm where more aqueous solutes
may interfere.
The o-tolidine method exhibits a sensitivity 1.6
times higher than the indigo method. Its non-specific
response to many kinds of oxidants, its low wave-
length region of absorption, and its temporal instabi-
lity, however, disqualify this method when compared
with the others (Sulzer, 1958; Hofman & Stern, 1969),
The DPD method for the determination of ozone is
based on a preliminary oxidation of iodide. However,
all methods based on oxidations of iodide to iodine
455
have the disadvantage of being non-specific; iodine is
also formed by secondary oxidants such as produced
by ozonation of water below pH 6-7 that contains
organic solutes. Moreover, the DPD method shows a
large time drift and therefore cannot be recommended
for extended series of measurements as required for
kinetic studies.
ratio of volume of reagent to volume of aqueous
ozone, the dynamic range is therefore restricted to a
factor of about 4.5.
In non-automated systems, preliminary tests will
help to locate the approximate range of concen-
tration before specifying the analytical procedure with
respect to ratios of volumes to be used. During our
studies, we covered a region of measurement of
0.005-30 mg l-1 of aqueous ozone by merely adjust-
ing the relative volume of aqueous ozone and of
Indigo Reagent applied. This entire range exhibits
constant sensitivity. It covers the requirements gener-
ally encountered in ozone applications.
4.5 Calibrations
In this study we used the u.v. absorbance of ozone
in distilled water at 258 nm as a secondary standard
to calibrate the analytical standard curves. At the
present time we still recommend such calibrations,
although different commercial products of indigo tri-
sulfonate tested so far have shown rather comparable
sensitivity factors (Hoign6 & Bader, 1980). Sensitivity
factors can, however, only be exchanged between
laboratories which perform the measurements on cali-
brated spectrophotometers at 600 nm. Filter instru-
ments will require individual calibrations.
5. CONCLUSIONS
The Indigo method covers the range of concen-
trations between 0.005 and 30 rag l-t. The method is
relatively straightforward and does not require any
difficult or time-consuming operations. It can be per-
formed with the classical instrumentation of water-
work laboratories. The Indigo Reagent is stable: it
can be stored for months as a reagent which is
"ready to use". The absorption to be measured at
600 nm is situated in a range where solutes of natural
waters do not interfere and where even simple visual
methods can be used for measurement. The change
of absorbance reflects the amount of ozone added
with high and constant sensitivity factor of AA =
- 2 0 , 0 0 0 c m - 1 ( m o l . l - l ) - I ozone added. All stan-
dard curves are linear. The sensitivity does not vary
with ozone concentration, with small changes of tem-
perature of reaction, or with the chemical compo-
sition of the water whenever the pH is below 4. For
many types of waters the instrumental or visual
456 H. BADERand J. HOI,GNF'~
measurements can be performed even a few hours
after the ozone has reacted with the Indigo Reagent.
The method is relatively selective. Secondary oxi-
dation products do generally not interfere. The reac-
tion of ozone with the Indigo Reagent is so fast that it
can be applied to stop and measure even very fast
ozonation reactions. The method has been well tested
and applied for the determination of ozone in many
different types of waters. Unexpected difficulties have
not arisen. More practical experiences with this
method are now being accumulated in many water-
works and laboratories.
However, this Indigo M e t h o d cannot be used as a
primary standard. At the present time the reagent
must still be calibrated. A standard curve can be pro-
duced using an aqueous solution of ozone in distilled
water which is calibrated by the direct u.v. absorption
measured at 258 nm or by the classical iodometric
procedure. It is, however, quite possible that a wider
application of the indigo m e t h o d will stimulate cen-
tral national laboratories or a chemical company to
market an indigo reagent with a calibrated sensitivity
factor for the determination of ozone and chlorine
dioxide (see Hoignb & Bader, 1980).
Acknowledgements--We thank Professor Werner Stumm
for his continuous encouragement, and Dr Jiirg Zobrist
and Dr Joan Davis for the valuable reviewing and dis-
cussions. Andreas Locher is thanked for his assistance in
performing many of the experiments during his apprentice-
ship stage.
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