Direct Colorimetric Method for the Determination

Of Chlorine Dioxide in Water

H. W. HODGDEN' and

R. S. INGOLS

Georgia Institute o f Technology, Engineering Experiment Station, Atlanta, G a .

0.4

HLORIXE dioxide v,-as suggested as a sterilizing agent for

water as early as 1900 by Berge ( 4 ) ,but it was not used on a
plant scale until 1944. I t is now used to some extent in water treatment for controlling tastes and for the removal of manganese
( 6 ) ,and in the textile, m-ood pulp, and paper industries for bleaching.
Methods currently used for the determination of the concentration of chlorine dioxide in water are basically the same as
those used for the determination of hypochlorous acid and the
chloramines. The relative merits of these methods of analysis
have been discussed by Ingols ( 7 ) . Two of the available methodi
(S, 6) are specific for chlorine dioxide but are indirect, while
others ( 2 , 9, I O ) are direct techniques-ie., one may determine
the concentration of chlorine dioxide in a pure solution-but
the techniques do not differentiate chlorine dioxide from hypochlorous acid in a mixture of the two. It would be highly desirable, therefore, t o develop a direct specific method for the determination of chlorine dioxide in water. Studies by Ingols et al.
(8) indicate that chlorine dioxide produces color when it is al-

0.3
U
W

f
0.2

P
Q

0.1

0
0.1
0.3
0.5
0.7
0.9
MOLAR RATIO OF TYROSINE TO CHLORINE DIOXIDE

Figure 2.

Effect of Increasing Tyrosine

Concentration

Constant 1.2 p.p.m. chlorine dioxide concentration a t room

temperature

Chlorine dioxide was generated from sodium chlorite with sulfuric acid. Chlorine-free, chlorine demand-free water was prepared by chlorinating a bicarbonate ion-buffered distilled water
and then dechlorinating with ultraviolet radiation. The sodium chlorite was received from the Mathieson Alkali Works,
Inc., and the tyrosine was purchased from the Eastman Kodak

co.

PREPARATION O F REAGENT BUFFER

POC

520

400

440

WAVE LENGTH

560

(m-1

Figure 1. Photospectrograph of Chlorine Dioxide-Tyrosine

Color Compound

Tyrosine (300 mg.) R-as dissolved in 100 ml. of O.L\- sodium

hydroxide, and then diluted t o 1 liter with distilled water containing 97 ml. of concentrated acetic acid and 150 grams of sodium
acetate. Reagent buffer must yield a final pH of 4.5 t o 4.6.

Taken with a B e c k m a n DU a t 5-mr intervals

lowed t o react with tyrosine, as was shown earlier by Aloy and

Rabault ( I ) . This paper presents the optimum pH and reagent
concentrations for the technique and an evaluation of reproducibility, sensitivity, and stability in the chlorine dioxide-tyrosine
color reaction.
METHOD O F RESEARCH

Chlorine dioxide was added t o a chlorine-free, chlorine demandfree water to give the desired concentration; this was determined by the method of Palin ( I O ) . Tyrosine reagent and a
buffer for pH control were added t o a portion of the sample and
the absorbance of the chlorine dioxide-tyrosine mixture was
determined by a Photovolt colorimeter Model 450 with a 150mm. light path and a B-490 filter with a maximum transmittance
at 490 mp. The choice of this filter is based on the spectrograph
of Figure 1. It was found that cobalt nitrate gave a similar
spectral absorption picture and that 140 p.p.m. of cobalt nitrate
was equivalent t o 0.1 p.p.m. of chlorine dioxide as chlorine.
Cobalt nitrate can be used as a permanent color standard for
field work with the chlorine dioxide-tyrosine reaction technique.
Studies were conducted a t temperatures of 22' to 24' C. and 4'
t o s o C.
1

Present address, Florida State Board of Health, Jacksonville, Fla.

RESULTS

The data as presented in Figure 2 indicate that the absorbance

of the chlorine dioxide-tyrosine compound reaches a maximum
at approximately 0.5 mole of tyrosine to 1 mole of chlorine dioxide
with no further change in absorbance between 0.5 and 0.8 mole
of tyrosine t o 1 mole of chlorine dioxide. Higher concentrations
of tyrosine cause no change in the color concentration.
The data as presented in Figure 3 indicate that there is an
increase in absorbance with a n increase of chlorine diovide in
ratios up to 2.0 moles of chlorine diovide to 1 mole of tyrosine
that is proportional t o the increase in concentration. From ratios
ranging from 2.0 t o 3.3 moles of chlorine dioxide to 1 mole of
tyro-ine there is a further increase in absorbance or color but it
IS not in proportion t o the increase in chlorine dioxide concentration. At ratios ranging from 3.3 to 8 moles of chlorine dioxide
to 1 mole of tyrosine, color is formed very rapidly but the colored
compound is further oxidized by the excess chlorine dioxide to
a colorless compound. The nature of chemical reaction is not
knonm, but probably the ring is broken and a simpler substituted
alanine is formed. It is important, therefore, that sufficient
tyrosine reagent be present when this method is used for the determination of chlorine dioxide.
1224

1225

V O L U M E 26, NO. 7, J U L Y 1 9 5 4
The apparent equation of the chlorine dioxide-tyrosine reaction which produces color is:
H H
C=C
2C102

+ HOC,\

H 0

irOH

C-C-C-

//

2C10;

+0

1 3

Tiris H y ~ i o c h l o r o u s.Icid.
2 ti

0.25
0.19
0.13

.4hscrLnnce
0.01
0 13
0 12
0.01
0.01
0 10

0.20
0.20
0.14

1.0

0.8
0.6

Tyro-ine Reagent, 111.

1
2
3

Chlorine
Dioxide,
P.P. M

HH

H H
C=C
C

H S

%-C
H H

Table I. Absorbance of Tyrosine Reagent with Chlorine

Dioxide with and without Hypochlorous Acid

0.23
0 19
0.13

1
3
P,P.AI.
7.0
____

0.0
0.0
0.0

0.0
0.0
0.0

Extinction coefficient640.

H H O

\
I!
C = C-C-COH

lngole et al. (S) reported that chlorine dioxide does not react

with the amino nitrogen atom of simple amino acid, rather, it

oxidizes the benzene of the phenol ring. Hypochlorous acid,
however, reacts with the nitrogen atom of amino acids and may
also react with the organic portion of the acid but by substitution
rather than oxidation.
The data presented i n Figure 4 indicate that a pH of 1.6 gives
the optimum absorbance in the chlorine dioxide-tyrosine reaction, while within the pH range of 4.3 to 5.2 only slightly less
color is produced. ThuF, within these pH values one ran detrrmine chlorine diovide by this method of analysis.
The data on reproducibility of this method of chlorine dioxide
determination, presented in Figure 5 , indicate that at the very
low concentrations of chlorine dioxide the coefficient of variation
(Cx = x X) shows that there is approximately a 12% variation

.I2

.IO

u_

!
>

-5

.M

>

.04

.02
0

0.5

1.0
p+m.

1.5
2.0
2.5
CHLORINE DIOXIDE

3.0

Figure 5. Relationship between

Chlorine Dioxide Concentration
and Best Reproducibility for 10
Replicates
Variability coefficient = rat constant pH,
X
temperature, and time of contact

MOLAR RATIO CHLORINE DIOXIDE TO TYROSINE

Figure 3. Effect of Increasing

Chlorine Dioxide Concentration
With a constant 3.0 p.p.m. tyrosine concentration in developing color at room
temperature

TIME MINUTES

Figure 6. Rate of Development and

Stability for Color
With 1.0 p.p.m. chlorine dioxide at pH 4.6 and
room temperature

45

5.0

5.5

60

PH

Figure 4.

Effect of Varying pH between 4 and 6 in

Color Formation

Rith 0.76 p.p.m. chlorine dioxide at room temperature with 10

minutes contest

in reproducibility even under the most carefully controlled conditions. At the higher concentrations of chlorine dioxide the probable error is 2 to 3% a t best.
A study was made to determine whether free chlorine would
increase the color of tyrosine and chlorine dioxide. The results,
shown in Table I, indicate not only that hypochlorous acid doe8
not increase the concentration of color, but that as the relative
concentration of the hypochlorous acid increases, it competes
for the tyrosine and prevents the formation of the colored compound unless an adequate amount of the tyrosine is present.
Because an oxidized manganese ion can react with o-tolidine
a t pH 2.0 to give a false chlorine residual, it was considered
possible that the oxidized manganese ions could produce a colored
tyrosine product. A natural water containing manganese was
used with and without both chlorine and chlorine dioxide as the

1226

ANALYTICAL

CHEMISTRY

warmed to 20' C. one should wait 8 to 10 minutes but not longer

than 15 minutes to compare or measure the color formed.

sample for testing for chlorine dioxide with the tyrosine reagent.
The chlorine plus manganese either in natural water or in a bicarbonate solution gives no color with the tyrosine reagent. The
chlorine dioxide oxidizes the manganous ions in the bicarbonate
solution of natural water instantly, causing a dark brolvn color
to form; when tyrosine reagent is added to the manganic solution
or suspension after treatment with chlorine dioxide, no tyrosinechlorine dioxide color develops.
The data of Figure 6 were obtained a t room temperature or
about 25" C. When the temperature of the chlorine diolide
solution was lowered to 4' C., 80 minutes were required to obtain the maximum color intensity.

CONCLUSIONS

Tyrosine is selective for chlorine diouide, producing a color

with which there is no interference from hypochlorous acid,
chloramines, and manganese. The method for the determination of chlorine dioxide is simple and rapid. The reproducibility
is adequate for higher concentrations. The sensitivity is poor
for the chlorine dioxide Concentrations less than 0.2 p.p.m. sometimes used in water treatment.
LITERATURE C I T E D

Aloy, XI. A1 , and Rabault, C., Bull. soc. chzm. Paris, Ser. 4,

RECOMMENDATIONS

111,391 (1908).
Am. Public Health Assoc., Xew York, "Standard AIethodn for
the Examination of Water and Sewage," 9th ed., 1946.
Aston, R. N., J . A m . Wuter Works Assoc , 42, 151 (1950).
Rerge, XI. A., MBm. sot. ing. civzls France, V 1900, Part I, B 475.
Haller, J. F., and Listek. S. C., AXLL.CHEM.,20, 639 (1949).
Harlock, C. R., Water & Sewage Works, 100, 74 (1953).
Ingols. R. S., J . Inst. Water Engrs., 4, 581 (1950).
Ingols, R. S.,Wyckoff, H. A., Kethley, T. .I.,Hodgden, H. W.,
Fincher, E., Hildebrand, J. C., and Mandel, J. E., Ind. Eng.
Chem., 45,998 (1953).
Marks, H. D., Williams, D. B., and Glasgow, G. G., J . Am.
Wuter Works Assoc., 43, 201 (1951).
Palin, A4.T., J . Inst. Water Engrs., 3, 100 (1949).

I t is recommended that 2 ml. of reagent be used per 100-ml.

sample for concentrations up to 2.5 p.p.m. of chlorine dioxide
plus hypochlorous acid and that with higher concentrations, the
sample be diluted below 2.5 p.p.m. or proportionately more reagent be used. Permanent cobalt nitrate standards can he
prepared for comparison of color produced in the chlorine dioxidetyrosine reaction. I t is recommended that cobalt nitrate standards contain 140 p.p.m. of cobalt nitrate for every 0.1 p.p,m.
of chlorine dioxide.
Because in the chlorine dioxide-tyrosine reaction the maximum
amount of color is produced to 6 in 8 minutes at 20" C. Ivhile
beyond 15 minutes the color begins to fade, after the solution is

RECEIVED
for review September 17, 19.53. Accepted March 18, 1954.
Investigations supported in part by a research grant from the National
Institutes of Health.

Detection of Bismuth by Dithizone in Molten Naphthalene

JACK K. CARLTON and WALTER C. BRADBURY
Institute o f Science and Technology, University o f Arkansas, Fayetteville, Ark.

HE organic reagent, dithizone (diphenylthiocarbazone), is

well known for itR use in the detection and determination of
certain metal ions (2). The reagent is commonly employed in a
chloroform or carbon tetrachloride solution and is used as an extraction medium for the desired metal ions. I n applying the
reagent, selectivity is attained by masking the interfering ions
with appropriate complexing agents, by the careful adjustment
of pH, or by a combination of these methods. I n certain cases
double extractions have proved advantageous; the metal ions
are extracted into the dithizone-chloroform medium and the
organic phase is then extracted by an aqueous solution the pH
of which has been adjusted to favor the solubility of the desired
metal dithizonate.
Feigl and Baumfeld ( 1 ) found that several metsls foinied
colored complexes in molten 8-quinolinol and have suggested
this as a method of detection for several of them. West and
Granatelli (3)studied the reaction of metal salts with molten
8-quinolinol microscopically and have established that the
characteristic crystals formed by a number of cations and anions
offer an extremely sensitive method of detecting them.
During an investigation of various reactions in nonaqueous
media the authors found that a solution of dithiaone in molten
naphthalene reacted v-ith bismuth salts very rapidly to yield a
brilliant red color. Bismuth concentrations in the range of
0.004 y are easily detectable. Reactions of the dithizone-naphthalene reagent with other metal ions and the nature of interferences and their masking were studied.
REAGENTS

Dithizone, Matheson Co., a 0.025% solution in Baker's

naphthalene.

6.p.

Bismuth chloride, J. T. Baker Chemical Co., prepared in 1 . O J I

hydrochloric acid to contain 1007 per drop. Dilutions were
made from this stock solution.
Reagent grade chemicals were used in the interference studies
and nere made up to contain 5007 per drop and were used in
this concentration. acid was added where needed to retain
hydrolyzable ions in solution.
EXPERIMENTAL

Because of the intense color of dithizone, maximum sensitivity

could not he obtained using this reagent undiluted in the molten
state. In addition, its melting point is relatively high and its
stability is lowered at higher temperatures. Consequently, naphthalene ivas selected as an inert, low melting solvent for the
reagent. The reagent solution was prepared by dissolving 16
nig. of dithizone in 75 ml. of chloroform, mixing with 66 grams
of naphthalene, and finally evaporating the chloroform from the
mixture.
Dilutions of the stock bismuth chloride solution were made
to establish the limit of identification of this ion. One drop of the
test solution was evaporated on a spot plate previously heated to
about 90" C. and a few milligrams of the reagent were added.
The red color of the bismuth dithizonate was clearly disrerned
a t a concentration of 0.0047 of bismuth. Limits of identification of other metal ions Rere determined following the same procedure. Observations of the reactions between various cations
and the reagent are listed in Table I. RIany of these gave salts
analogous to those obtained employing the chloroform solution
of dithizone, while others vary considerably. In some cases the
color changes required long periods of time and relatively large
concentrations of the metal ion. Cadmium, tin, mercury, zinc.
and silver gave almost instantaneous reactions.