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Analyst, January, 1969, Vol. 94, @. 43-48 43

The Determination of Sulphur Dioxide with

Rosaniline Dyes
BY H. G. C . KING AND G. PRUDEN
(Rothamsted Experimental Station, Harpenden, Herts.)

The compositions of commercial rosaniline hydrochloride (magenta) and

pararosaniline hydrochloride samples have been re-investigated to improve
the colorimetric determination of sulphur dioxide. The dyes, purified in small
amounts by paper chromatography, each give a linear calibration up to
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043

-
80 pg of sulphur dioxide, with the hydrochloric acid formaldehyde reaction
at pH 1.1. Of the two dyes, pararosaniline hydrochloride is preferred because
of its smaller reagent blank value. Impurities can be readily removed from
pararosaniline base by recrystallisation, and this reagent is, therefore,
recommended for routine work.
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INseeking methods to improve the colorimetric finish to the determination of sulphur dioxide
from sulphur in soils by Bloomfield's vanadium pentoxide method,l we have re-investigated
the relative merits of pararosaniline hydrochloride (Ia) and rosaniline hydrochloride (Ib,
magenta or basic fuchsin) as colorimetric reagents in the sulphite-hydrochloric acid-
formaldehyde reaction.

c1-

NH2

la (R=H), Pararosaniline hydrochloride

Ib (R=CH,), Rosaniline hydrochloride (magenta)

Acid-bleached magenta, first used by Steigmann2as a quantitative reagent for bisulphite,

was later used by Urone, Boggs and Noyes3 to determine sulphur dioxide in air. The use
of magenta was largely discontinued about 6 years after its introduction as a quantitative
reagent, because of an alleged lack of purity, usually ascribed to admixed paarosaniline hydro-
chloride and to other, unspecified, compounds. It is known that pararosaniline hydrochloride
may be formed during the manufacture of magenta, although the cause of erratic results,
which some authors attribute to the impurities, has not been shown conclusively to be the
result of the use of mixed magenta - pararosaniline hydrochloride reagents.
The substitution of pararosaniline hydrochloride for magenta was first made by West
and Gaeke? who did not discuss the purity of the new reagent. Later workers,6s6 however,
drew attention to the existence of a variety of commercial pararosaniline reagents of varying
dye potency. Pate, Lodge and Wartburg6 assayed spectrophotometrically eighteen samples
of pararosaniline hydrochloride and basic fuchsin, and accepted only those showing the
spectral maximum of the purest pararosaniline then available. More recently Scaringelli,
Saltzman and Frey7 made a detailed study of the conditions affecting the determination of
sulphur dioxide colorirnetrically, and used a counter-current distribution procedure to purify
pararosaniline hydrochloride.
We have examined various samples of pararosaniline hydrochloride and magenta by
paper chromatography, and have further obtained sufficient of each of the pure dyes by
0SAC and the authors.
 44 KING AND PRUDEN: THE DETERMINATION OF [AIzalyst,
View Article VOl.
Online94
chromatography on thick paper to compare their usefulness in the colorimetric determination
of sulphur dioxide. Standard sulphite solutions were stabilised as the disulphitomercurate
ion: [Hg(SO,),]2-, and the colour of the complex with the acid-bleached dyes was deter-
mined at pH 1-1 after reaction with formaldehyde. With either dye a linear calibration
was found from 0 to 80 pg of sulphur dioxide, but the reagent blank (E = 0.024) for para-
rosaniline hydrochloride at the wavelength of maximum optical density (572 nm) was about
one tenth of that (E = 0.196) of magenta at the same wavelength.
Although none of the magenta samples we examined contained pararosaniline hydro-
chloride as an impurity, it is possible that there are such samples, the purification of which
would be difficult on a large scale.
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043

Also, because of its small reagent blank pararosaniline hydrochloride would be preferred
as a reagent for determining sulphur dioxide. However, the preparation of the pure dye on
a small scale by paper chromatography is unsuitable for routine work, especially if the dye
contains a large proportion of impurity.
We find that pararosaniline base (11)-
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NH2

II, Pararosaniline base

prepared by treating an acid-bleached solution of the hydrochloride with sodium hydroxide,

can be substituted with advantage for the parent dye. The base can be made on as large
a scale as convenient and, when recrystallised from aqueous methanol, is a reproducibly pure
and stable reagent. Variability in the quality of the pararosaniline hydrochloride is over-
come by removing impurities when the base is recrystallised. Rosaniline base, although it
can be prepared and recrystallised in the same way as pararosaniline base, suffers from the
disadvantage of its parent dye in giving the same high blank value in the determination.
EXPERIMENTAL
PARAROSANILINE HYDROCHLORIDE-
After a chromatographic and spectrophotometric examination of various samples, one
that contained few impurities (Hopkin & Williams Ltd., Catalogue No. 6496.6) was selected
for the preparation of small amounts of the pure dye by preparative paper chromatography
and for making pararosaniline base.
MAGENTA-
Samples, some many years old, from various sources were examined. No sample was
found to contain pararosaniline hydrochloride as an impurity, although one, labelled Basic
fuchsine, consisted almost entirely of pararosaniline hydrochloride, with no magenta. In-
organic salts were absent from every dye sample, but all of the samples contained more or
less organic impurity. Small amounts of the pure dye could readily be isolated by paper
chromatography.
A sample of magenta, For the preparation of Schiffs reagent (British Drug Houses
Ltd.), was selected for preparative paper chromatography.
METHODS
PAPERCHROMATOGRAPHY-
From the many combinations of solvents tested, the simplest solvent that would separate
pararosaniline hydrochloride and magenta, giving appreciable RF values, was 60 per cent.
 January, 19691 SULPHUR DIOXIDE WITH ROSANILINE DYES View Article Online45
aqueous ethanol. Solutions of pararosaniline hydrochloride and magenta (0.5 per cent.
w/v in methanol) were applied to Whatman No. 3 chromatographic paper in the form of
bands about 2 cm long, by using a fine glass capillary. The use of thick paper, rather than
the thinner Whatman No. 1 and No. 2 grades, helps to keep the bands as compact as possible
during the run. At a temperature of 22" C the solvent front travelled 30 cm (descending)
in 64 hours, after which time the bands had increased to about three times their starting
width. RF values, to the front edge of the bands, were 0.78 for pararosaniline hydrochloride
and 0.83 for magenta. Although mixtures of the dyes could be distinguishedby their colours,
definite separation of the bands was made difficult by their broadening during the run.
Each dye contained a violet-coloured mobile impurity, RF value 0-70. Magenta samples
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043

also contained a second violet impurity that ran immediately in front of the main dye band.
Each dye left at the origin a blue - violet band that could not subsequently be eluted. The
mobile bands could all be eluted with methanol.
For preparative work the dyes were banded on to several Whatman No. 3 papers, 25 em
wide, cut to a point at their lower edge. The main dye bands were collected in beakers
placed under the papers during prolonged elution with the chromatographic solvent.
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SPECTROPHOTOMETRY-
Measurements of spectra in the visible region were made in 1-cm cells with a Hilger
& Watts Spectrochem Mark 2 spectrophotometer. Purified pararosaniline hydrochloride and
magenta were dissolved in water or ethanol and diluted to give a final concentration of 0.5 mg
in 100 ml of solvent. Pararosaniline base, in aqueous suspension, was made just acid with
hydrochloric acid to regenerate the hydrochloride, then diluted with water to give the same
final concentration. A solution of the base in methanol, in which it is sparingly soluble was
diluted to give a final concentration of 0.5 mg per 100 ml of methanol.

Wavelength, nm

Fig. 1. Visible spectra of A,

pararosaniline hydrochloride in water ;
B, magenta in water; and C, para-
rosaniline base in ethanol

The wavelength of maximum optical density of pararosaniline hydrochloride was 536 nm

in water and M5nm in ethanol. For magenta, the maxima were 540nm in water and
546 to 547 nm in ethanol. Pxarosaniline base, which is insoluble in water, showed a maxi-
mum at 541 nm in ethanol. Fig. 1 shows the spectra of the dyes in water and of the base
in ethanol.
 46 KING AND PRUDEN: THE DETERMINATION OF [Afldyst,
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Article VOl.
The violet impurity, RF value 0-70, showed a maximum at 639 nm for water, and at
fj49nm for ethanol. The fast-running impurity in magenta had maxima at MOnm for
water and 649 nm for ethanol. None of the impurities, in butanol, showed a maximum as
high as 660 nm, reported by Scaringelli, Saltzman and Frey.7
PREPARATION OF PARAROSANILINE BASE-
One gram of pararosaniline hydrochloride was dissolved in 250 ml of 2.6 N hydrochloric
acid. After standing for 2 hours the bleached solution was filtered through a Whatman No. 1
filter-paper. The base was precipitated as pale magenta-coloured shiny plates by adding
a slight excess of 2 . 5 ~sodium hydroxide solution to the filtrate, and the precipitate was
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043

collected on a sintered-glass plate, porosity 3; the filtrate was colourless. The crude base
was washed thoroughly with water to remove sodium chloride and excess of sodium hydroxide,
and was then recrystallised by dissolving in 70 ml of methanol at the boiling-point, adding
300 ml of water at 80" C and allowing the solution to cool at room temperature. The re-
crystallised product, 64 per cent. of the weight of starting material, blackened in the range
200" to 205" C and decomposed at 286" C. Scaringelli, Saltzman and Frey7 prepared the
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base by recrystallisation from water as an intermediate in their subsequent preparation of

pararosaniline hydrochloride. We found that recrystallisation from water gave a yield of
only 12 per cent.
Recrystallisation from aqueous methanol undoubtedly leads to some loss of product,
which is retained in the filtrate, but impurities that are soluble in the solvent are completely
removed, as shown by a chromatographic check on the hydrochloride prepared from the pure
base.
COLORIMETRIC DETERMINATION OF SULPHUR DIOXIDE-
Bleached 9ararosaniline reagent solutiort-One gram of pararosaniline base is dissolved
in 60ml of concentrated hydrochloric acid and the solution diluted to 1 litre with water.
We have not had the opportunity to study the stability of the reagent solution over a long
period, but have kept a solution unchanged for 6 weeks in the dark; the reagent solution of
Scaringelli, Saltzman and Frey' is reported to be stable for at least 9 months.
Potassium tetrachZoromercurate solutim-A solution of 27.2 g of mercury(11) chloride and
14-9g of potassium chloride in water is diluted to 1 litre.
Formaldehyde solution, 0.2 per cent.-Five millilitres of 40 per cent. formaldehyde solution
are diluted to 1 litre with water. The solution should be freshly prepared.
Standard sulphite solution-A solution of 0-4 g of anhydrous sodium sulphite in 500 ml
of water is prepared, corresponding to between 300 and 400pg of sulphur dioxide per ml.
The concentration of sulphite is determined iodimetrically. One millilitre of 0-1 N iodine
solution is equivalent to 3.2033 mg of sulphur dioxide.
Immediately after analysis the sulphite solution is stabilised by dilution with potassium
tetrachloromercurate solution. Ten millilitres of sulphite solution diluted to 600 ml with the
tetrachloromercurate give a solution corresponding to between 6 and 8 pg of sulphur dioxide
per ml.

Wavelength, nm
Fig. 2. Visible spectrum of the sulphur dioxide
-
(60 p g ) pararosaniline hydrochloride - formaldehyde
complex
 January, 19691 SULPHUR DIOXIDE WITH ROSANILINE DYES View Article Online47
CALIBRATION-
Aliquots of the dilute sulphite solution, corresponding to not more than 80 pg of sulphur
dioxide, are transferred to 50-ml graduated flasks, and 10 ml each of bleached pararosaniline
reagent and formaldehyde are added. The solutions are diluted to 50 ml (final pH 1-1)and
are then allowed to stand for 20 minutes before measuring the optical densities, in 1-cm glass
cells, at 672 nm against a reagent blank.
Fig. 2 shows the visible absorption spectrum for the developed colour, and Fig. 3 the
calibration graph.
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043
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Sulphur dioxide, mg
Fig. 3. Calibration for sulphur dioxide with A, para-
rosaniline hydrochloride; and B, magenta, against respective
reagent blanks

DISCUSSION
In the various dye samples we examined, neither the amounts nor the nature of the
impurities were such as to give variable results in the colorimetric determination of sulphur
dioxide. It is difficult to compare our results directly with those of other workers, who used
reagents of varying quality from many sources but, unless objection is made to the higher
reagent-blank value of magenta, there seems to be no reason why either of the rosaniline dyes
should not be used. Although none of our magenta samples was contaminated with para-
rosaniline hydrochloride, we found that one sample, labelled Magenta, was pararosaniline
hydrochloride, and that one sample of the latter was magenta.*
If pararosaniline hydrochloride contains a large amount of impurities, its optical density
will be much less than Ei& = 2600 at 536 nm (water), but small amounts of the pure com-
pound can be obtained by preparative paper chromatography. Column chromatography on
cellulose is unsatisfactory because the dye is adsorbed strongly on the cellulose; this does
not happen on paper chromatograms unless the paper is dried thoroughly after a run and
the chromatogram is then re-run. For gram-scale amounts of the reagent when only impure
samples are available, or as a routine measure, it is preferable to use pararosWne base,
which can be prepared in good yield as a pure, crystalline product. The base gives an extremely
pale acid-bleached reagent solution and as good a calibration as the pure hydrochloride.
The impurities in pararosaniline hydrochloride and magenta can be readily detected by
the chromatographic system described above. The small difference in the structures of the
dyes makes it more difficult to detect a mixture, but this can be done if the bands of dye
applied to the starting line of the chromatogram are kept as narrow as possible. It is useful,
but not always necessary, just to dry the chromatogram and re-run it in the same solvent
to improve the separation.
Much information about possible contamination of magenta with pararosaniline hydro-
chloride can be gained by a study of the spectral maxima of mixtures of the dyes. The
spectral assay by Pate, Lodge and Wartburg6 showed that it is possible to distinguish between
the dyes; the grouped samples having a maximum optical density at 543 nm as para-
rosaniline hydrochloride, and those with maximum at 549 nm were grouped as basic fuchsin
* Large blank values were found by Bloomfield in the colorimetric finish to his determination of sulphur
in soils1 (Dr. C. Bloomfield, personal communication). His pararosaniline hydrochloride was, in fact,
magenta.
 48 KING AND PRUDEN View Article Online
(magenta). Pate, Lodge and Wartburg do not state specifically the nature of the solvent
in which measurements were made, and we have found no solvent for which the maximum
optical density of magenta is as high as 549 nm.
Earlier workers failed to point out the importance in spectral assay of the lower optical
density of magenta compared with that of an equal concentration of pararosaniline hydro-
chloride. Magenta that contains more than 10 per cent. of pararosaniline hydrochloride
shows a decrease in its wavelength of maximum optical density, accompanied by an increase
in its original optical density. For a spectral assay it is necessary first to prepare small
amounts of the pure dyes chromatographically.
Table I shows the change in optical density and maximum wavelength of mixtures of
Published on 01 January 1969 on http://pubs.rsc.org | doi:10.1039/AN9699400043

the dyes in water.

TABLEI
CHANGESIN SPECTRAL MAXIMUM AND OPTICAL DENSITY OF MIXTURES OF MAGENTA
AND PARAROSANILINE HYDROCHLORIDE AT A TOTAL CONCENTRATION OF
0 6 m g IN 1 o o d O F WATER
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Pararosaniline, % 0 10 20 30 40 60 60 70 80 90 100
Magenta, % ,. 100 90 80 70 60 60 40 30 20 10 0
Emax.(nm) .. 640 640 639 639 638 638 637 637 637 636 636
0.889 0.909 0.960 0.960 0.984 1.107 1.177 1.180 1.193 1.293 1-303

Variations in the conditions for the colorimetric determination of sulphur dioxide have
been studied by Scaringelli, Saltzman and Frey,' who measured the developed colour at one
of two pH values, 1.6 (Method A), Amax. = 548nm, giving a large reagent blank value, or
at 1.2 (Method B), Amax. = 575 nm, giving a smaller reagent blank, the change from the
higher to the lower pH value being made by adding 3 N orthophosphoric acid. Under our
conditions, with 100 mg of pararosaniline base in 1 litre of 6 per cent. v/v hydrochloric acid,
the pH of the reaction mixture is 1.1 and the wavelength of maximum optical density is
572 nm.
We suggest that, to eliminate variations in the determination of sulphur dioxide when
impure commercial pararosaniline hydrochloride has been used, recrystallised pararosaniline
base should be substituted.
A considerable improvement can be made at once to the determination of sulphur in
soils (measured as sulphur dioxide) to improve the original calibration of 2 to 25 pg of sulphur,
linear in the range 0 to 10pg.l The determination of sulphur dioxide from other sources,
e.g., from air, or in foods and drinks, may require modifications to the initial stages of the
method to inactivate interfering substances, e.g., heavy metals, nitrogen dioxide and ozone.
These modifications are fully discussed by Scaringelli, Saltzman and Frey,' but the final
colorimetric determination of sulphur dioxide is unaltered.
It has been suggested8 that p-aminoazobenzene is a superior reagent to pararosaniline
hydrochloride. We cannot confirm this ;the reagent blank for the 9-aminoazobenzene reagent
has twice the optical density of that of magenta, and the calibration is linear only up to
60 pg of sulphur dioxide.
REFERENCES
1. Bloomfield, C., Analyst, 1962, 87, 686.
2. Steigmann, A., Analyt. Chem., 1950,22, 492.
3. Urone, P., Boggs, W. E., and Noyes, C. M., Ibid., 1961, 23, 1617.
4. West, P.W., and Gaeke, G. C., Ibid., 1956, 28, 1816.
6. Pate, J. B., Lodge, J. P., and Wartburg, A. F., Ibid., 1962, 34, 1660.
6. Barabas, S., and Kaminski, J., Ibid., 1963, 35, 1702.
7. Scaringelli, F. P., Saltzman, B. E., and Frey, S. A., Ibid., 1967, 39, 1709.
8. Kniseley, S. J., and Throop, L. J., Ibid., 1966, 38, 1270.
Received August lst, 1968