J. Soc. Cosinefir Chemists 19 213-223 (1968) lo68 Socuty o!

Cosmetic Chemistt of Great Britain

Estimation of hexachlorophane
cosmetic products
in
A REPORT BY THE METHODS OF ANALYSIS
SUBCOMMITTEE OF THE SOCIETY OF
COSMETIC CHEMISTS OF GREAT BRITAIN
Syn0psisMethods for analysing hexachlorophane in toiletries and cosmetics are reviewed,
and discussed. Preliminary experiments with uv absorption methods indicated that the differ-
ential absorption method was preferable to the one measuring absorption at fixed wave-
lengths, which could suffer interference from extraneous and irrelevant absorption.
Interlaboratory collaborative tests have been carried out on certain toilet preparations,
viz. alcoholic solutions, talcs and soaps, containing hexachlorophane at levels between 0.2
and 2 o, using the differential absorption method of Elvidge and Peutrell. Results show that
the method is sufficiently accurate and specific for the determination of hexachlorophane in
these preparations.
INTRODUCTION
Hexachlorophane is used in a large number of cosmetic and toiletry
products. It functions as a germicide, bactericide or deodorant in products
such as talcs, soaps, deodorant and/or antiperspirant preparations, tooth-
pastes, etc., and is generally used at a concentration between 0.01% and
2.0%.
The Methods of Analysis Subcommittee of the Society of Cosmetic
Chemists of Great Britain has reviewed existing methods for estimating
hexachlorophane to find a simple specific method suitable for estimating
this material in toilet preparations.
REVIEW OF POSSIBLE METHODS
Published methods can be grouped as follows:
(1) Titration methods.
213
214 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS
(2) Colorimetric methods.
(3) Measurement of absorption at specific wavelengths.
(4) Differential absorption measurements.
(5) Gas chromatography.
(6) Determination of organic chlorine.
Titration methods
Potentiometric titration (1) is suitable for the pure material but not
for hexachlorophane in a product. High frequency titration techniques (2)
have been used for various phenols but they require special equipment.
Colorimetric methods
Several colorimetric methods have been used.
t Amino antipyrine-potassium ferricyanide
This method is widely used and was first published by Gottlieb and
Marsh (3). Mohler and Jacob (4) considered it to be the most sensitive and
accurate method available, whereas Larson (5) and Lord, McAdam and
Jones (6) reported that results were unreliable. The sensitivity of the
reaction is pH dependent and the result is not specific for hexachlorophane.
Achmeteli (7) overcame the interference from methyl salicylate in
toothpaste by using methylene blue instead of potassium ferricyanide, but
anionic detergents also react with methylene blue and must first be removed.
2.6 Dibromoquinone chloroimide
This method, often known as the Gibbs method (8), is based on the
formation of a dibromo-indophenol dye which can be estimated colori-
metrically. It has been adapted by Singer and Stern (9) for the determina-
tion of bisphenols such as hexachlorophane, but is not specific.
Nitrosophenol
Gibbs (10) examined the reaction of phenols with nitrous acid and the
[ormation of coloured nitroso compounds. Stroughton (11) developed a
method for the preparation of nitroso compounds with nitric and sulphuric
acids at 100C, and their rearrangement in the presence of excess alcoholic
ammonium hydroxide to produce a highly coloured quinonoid radical.
Lykken, Treseder and Zahn (12) modified this method and used sodium
nitrite in acetic acid to form the nitrosophenol at room temperature.
ESTIMATION OF HEXACHLOROPHANE IN COSMETIC PRODUCTS 215
Ferric chloride
Larson (5) used this reagent to determine hexachlorophane in soaps.
Most of the colorimetric methods were originally developed for simple
phenols and have to be adapted for chlorinated bisphenols. In general they
are nonspecific, although individual phenols do give slightly different colours.
Measurement of absorption at specific wavelengths
Lord et al (6) measured the uv absorption of bisphenols in alkaline
solution and based a method of determination on well defined maxima in
the region 300-330 nm. The absorption is determined at three wavelengths,
and a Morton and Stubbs (13) correction is applied to allow for irrelevant
absorption due to soap. Clements and Newburger (14) quoted methods for
extracting bisphenols from soaps and cosmetic preparations, and determined
the amount present by measuring the uv absorption in alcoholic solution
at three specified wavelengths and applying appropriate corrections.
Differential absorption measurements
Mahler (15) reported that the uv absorbance of bisphenols varies with
the state of ionisation. Childs and Parks (16) used this phenomenon to
determine hexachlorophane in liquid soap by measuring the difference in
absorption at two pH values.
Elvidge and Peutrell (17) modified the pH 8 buffer by replacing sodium
hydroxide in methanol with trishydroxymethyl amino methane. The pH 8
buffer was selected because chlorinated bisphenols are fully ionized at this
pH but phenol, cresol and resorcinol are not, thereby reducing interference.
Elvidge and Peutrell also gave extraction methods for various types of
cosmetic preparations.
Gas chromatography
Porcaro (18) estimated dichlorophane, hexachlorophane and other
bisphenols by glc, using short columns (0.2-0.3 m), a temperature of 200-
250C and fast flow rates. The carrier gas was helium and the stationary
phase DC 710 (phenyl methyl silicone); under these conditions retention
times were between 5 and 15 min and the sensitivity 5- 10 gg.
Determination of organically combined chlorine
Any of the standard methods can be used but the preferred method is
216
JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS
probably a Sch6niger type combustion. This procedure cannot differentiate
between hexachlorophane and other organic compounds containing chlor-
ine.
CONSIDERATION OF METHODS
In considering these methods it was borne in mind that the hexachloro-
phane would possibly be present in a toilet or cosmetic product at low levels
and that only limited equipment may be available. Other phenolic materials
could also be present, hence it was desirable to have a specific method.
All the titration methods are non-specific and therefore were not
considered further. Likewise the colorimetric methods are non-specific
since the differences between the colours produced by the various phenols
are often not sufficient for characterisation.
After some of the work was started a gas chromatographic method (18)
was published, but it was decided not to do any practical work because
some members of the subcommittee did not have suitable facilities.
Nevertheless it is a specific method, which is relatively simple if the equip-
ment is available.
Determination of organic chlorine is lengthy and non-specific.
Measurement of uv absorption at specific wavelengths with a correction
to allow for irrelevant absorption, and the differential absorption method,
which makes an allowance for absorption by other phenols appeared worthy
of further attention.
They were examined using typical cosmetic products containing
hexachlorophane.
INITIAL EXPERIMENTS
Samples of an alcoholic solution, a deodorant stick, a talc and a soap
each containing hexachlorophane, were distributed amongst six laborator-
ies. Each sample was examined for its hexachlorophane content by two
methods:--
(1) Absorption at three specified wavelengths (14), and
(2) differential absorption (17).
Both papers also describe 'the preparation of samples and these methods
were used where appropriate.
The results for the products are summarised in Table I.
ESTIMATION OF HEXACHLOROPHANE IN COSMETIC PRODUCTS 217
218 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS
The first sample of soap had been hand-prepared, and the hexachloro-
phane had not been uniformly dispersed, so that the results showed a
rather wide variation. These results were discarded and instead a commercial
soap was examined by the differential absorption method only, when more
uniform results were obtained.
The difference between the results obtained by the two methods is
marginal although the differential method perhaps tends to give slightly
lower results. However, members preferred the latter method and results
were slightly less variable. Two laboratories carried out tests, and found
that the method was quantitative for pure hexachlorophane.
STANDARDISATION OF METHOD
A discussion at this point showed that there had been minor variations
in the preparation of samples. The method was therefore carefully edited
to avoid any further ambiguity, and details are given in the Appendix.
THE EFFECT OF PERFUME CONTAINING PHENOLIC CONSTITUENTS
Two alcoholic solutions were tested, one without perfume and the other
with a relatively high level of a number of perfumery compounds which
could conceivably interfere.
The results are given in Table II.
Table II
No. of No. of mean range
laboratories results
--
Non-perfumed solution
0.20 % w/w hexachlorophane 6 8 0.197 o 0.19-0.20
Perfumed solution
0.20 % w/w hexachlorophane
and 0.$ perfume 6 8 0.200 0.19-0.22
Composition of perfume
Parts
Bergamot oil 50
Amyl salicylate 20
Exalavende 10
Cananga oil 5
Orantiol 5
Clove oil 5
Coumarin 3
Birch tar oil 2
Musk D.T.I. 2
102
The results indicated, and subsequent statistical analysis confirmed,
that interference from perfume could be neglected.
ESTIMATION OF HEXACHLOROPHANE IN COSMETIC PRODUCTS 21,0
FURTIlER EXPERIMENTS AND STATISTICAL ANALYSIS
Homogeneous samples of soap and talc containing hexachlorophane
were distributed to participating laboratories and the results obtained using
the standardised method are given in Table III.
Table III
Labora- Soap (0.40 %) Talc (0.35 %)
-i-
tory 1 2 3 1 4
0.37
0.37
0.38
0.31
0.40
0.38
0.33
0.38
O.4O O.39
0.38 0.38
0,35 0.38
0.35
0.35
0.36
0.28
0.33
0.35
2 3
0.35 0.35
0.34
0.34 0.34
0.37 0.35
0.35
0.34
0.34
The results from laboratory 4 were definitely low, significantly so in the
case of the talc, owing to a fault in the spectrophotometer. These results
were therefore excluded from the statistical analysis (Table IV).
Table IV
Statistical analysis
Mean Range Standard 95 % confidence
deviation limits
Soap 0.40 w/v 0.38 0.33-0.40% 0.0210 +_0.045
hexachlorophan e
Talc 0.35 % w/w 0.35 0.33-0.37 % 0.0100 +_0.021
hexachlorophane
Alcoholic solution 0.20 0.19-0.22 0.0074 _+ 0.016
0.20 % w/w
hexachlorophane [
SPECIFICITY OF THE METHOD
Other chlorinated bisphenols which might be present in cosmetic
preparations are dichlorophane-2,2' methylene bis (4 chlorophenol) and
bithionol-2,2' thio bis (4,6 dichlorophenol).
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JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS
The differential absorption curves for these two compounds were
compared with that of hexachlorophane (Fig. 1 and Table V).
Table V
Spectroscopic data
ttexachlorophane
Dichlorophane
Bithionol
inx nm
312
307
327
max cm- 1
32,050
32,600
30,500
approx. E 1
lcm
144
32
157
160 -
140 --
100
80
60
4O
2O
30,500 cm-I
327 nm
32,050 cm-I/["X
312 nm , ,
/ \
32,600cm?
307nm J
, \
\
,,
i \\\
Hexachlorophane
.... Dichlorophane
Bi'l'hionol
o o
o o g o g o o o o o
trigur i Differential absorption spectra
ESTIMATION OF HEXACHLOROPHANE IN COSMETIC PRODUCTS 221
The curve for bithionol is sufficiently different from that of hexachloro-
phane to ensure that it will not intefere. The maxima are much closer for
hexachlorophane and dichlorophane but if there is a mixture the maximum
absorption lies between 307 and 312 nm according to the amounts present,
e.g. a 1:1 mixture has a maximum absorption at 310 nm.
These results indicate that dichlorophane and bithionol will be detected,
should these compounds be present.
DISCUSSION
The results were generally satisfactory and no account was taken of the
type of spectrophotometer used in the collaborative tests. It was found
necessary to standardise the procedure fairly tightly to secure comparable
results. The addition of likely perfumery compounds showed that there was
no significant interference from this source.
For the soap sample there was a tendency for the results to be a little
lower than expected. However, recovery of hexachlorophane added to
untreated soap chips was quantitative and it is considered that the method
is satisfactory. A different extraction procedure may be necessary for other
types of product containing hexachlorophane, e.g. antiperspirant/deodorant
products and toothpastes.
CONCLUSIONS
Inter-laboratory collaborative tests have shown that the differential
absorption spectrophotometric method published by Elvidge and Peutrell
is sufficiently accurate and specific for the determination of hexachlorophane
in certain toilet preparations, e.g. alcoholic solutions, talcs and soaps,
containing hexachlorophane levels between 0.2% and 2%.
ACKNOWLEDGEMENTS
The Committee are grateful to the following companies and organisa-
tions who allowed their staff time and facilities to take part in the collabora-
tive work:--Beecham Products U.K., Chesebrough-Pond's Ltd., Goya Ltd.,
Givaudan & Co. Ltd., Huntingdon Research Centre, The Pharmaceutical
Society of Great Britain, and Unilever Research Laboratory- Isleworth.
(Received: 18th December 1967.)
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JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS
REFERENCES
(1) Technical bulletin H 7. Sindar Corporation, U.S.A.
(2) Ershov, B. P. et al., Khim. Prom. 106 (1957); Anal. Abstr. 5 1904 (1958).
(3) Gottlieb, S. and Marsh, P. B. Ind. Eng. Chem. Anal. Edition 18 16 (1946).
(4) Mohler, E. F. and Jacob, L. N. Anal. Chem. 29 1369 (1957).
(5) Larson, H. L. J. An. Oil Chemists Soc. 28 301 (1951).
(6) Lord, J. W., McAdam, I. A. and Jones, E. B. Soap Perfumery Cosmetics 26 783 (1953).
(7) Achmeteli, H. I. J. Assoc. Offic. Agr. Chemists 48 278 (1960).
(8) Gibbs, H. D. J. Biol. Chem. 72 649 (1927).
(9) Singer, A. J. and Stern, G. R. Anal. Chem. 28 1511 (1951).
(10) Gibbs, H. D. J. Biol. Chem. 71 445 (1927).
(11) Stroughton, R. W. J. Biol. Chem. 115 293 (1936).
(12) Lykken, L., Treseder, R. S. and Zahn, V. Ind. Eng. Chem. Anal. Edition 18 103 (1946).
(13) Morton, R. A. and Stubbs, A. L. Analyst 71 348 (1946).
(14) Clements, J. E. and Newburger, S. H. J. Assoc. Oc. Agr. Chemists 117 190 (1954).
(15) Mahler, W. J. Am. Chem. Soc. 76 3920 (1954).
(16) Childs, R. F. and Parks, L. M. J. Am. Pharm. Assoc. Sci. Ed. 45 313 (1956).
(17) Elvidgc, D. A. and Peutrell, B. J. Pharm. Pharmacol. 13 Suppl. 111T (1961).
(18) Porcaro, P. J. Anal. Chem. 36 1664 (1964).
ESTIMATION OF HEXACHLOROPHANE IN COSMETIC PRODUCTS 223
Appendix
DETERMINATION OF HEXACHLOROPHANE BY DIFFERENTIAL ABSORPTION
Principle
Hexachlorophane in solution at pH 3 (unionised) has a lower extinction value than at
pH 8 {ionised), whereas the spectra of most other constituents of the toilet preparation are
unchanged. Tlxe hexachlorophane content is determined by measuring the difference in
extinction which has a maximum at 312 nm.
Reagents
pH 8 buffer:
Dissolve 6.07gtris-(lxydroxy-methyl) aminomethane (2 amino 2-(hydroxy-methyl) propane
-1:3-diol) in 900 ml methanol. Add 50 ml 0.SN hydrochloric and make up to 1 1 with distilled
water. Nix thoroughly.
ptt 1.4 buffer:
Add 18 ml glacial acetic acid and 40 ml N hydrochloric acid to 900 ml methanol. Dilute
to 1 1 with distilled water and mix thoroughly.
Preparation of samples
Soap:
Accurately weigh about 2g of soap into a 100 ml volumetric flask. Add 10 ml water and
warm gently on a water bath until dispersed (1 ml of pH 8 buffer may,be added if necessary
to prevent excessive foaming). Dilute to 100 ml with pH 8 buffer keeping the solution at
30-35C. Dilute one 10 ml aliquot to 50 ml with pH 8 buffer and a second 10 ml aliquot to
50 ml with pH 1.4 buffer keeping the temperature between 30 and 35C. The solutions at this
dilution should be clear and will not gel for at least 1 hr even if cooled to room temperature.
Proceed with spectrophotometric determination as soon as possible.
Talc:
Accurately weigh about 2g of talcum powder into a No. 3 porosity, sintered glass crucible
or funnel. Extract the powder with 5 successive portions of 20 ml chloroform, draw each
extract through the sinter with gentle suction and collect the combined extracts in a Buchner
flask. Transfer to a beaker and remove the chloroform on a steam bath, taking care to avoid
volatilisation of the bisphenol. This is best achieved by evaporating almost to dryness, then
removing the beaker from the bath and finishing the evaporation using a jet of compressed
air. Dissolve the residue in pH 8 buffer and dilute to 100 mi. Dilute one 10 ml aliquot to 50ml
with pH 8 buffer and a second 10 ml aliquot to 50 ml with pH 1.4 buffer. Proceed with the
spectrophotometric determination.
Alcoholic solution:
Pipette 10 ml of the alcoholic solution into a 100 ml graduated flask and dilute to volume
with pH 8 buffer. Dilute one 10 ml aliquot to 50 ml with pH 8 buffer and a second 10 ml
aliquot to 50 ml with pH 1.4 buffer.
If a w/w result is required, determine the density of the original solution or weigh a 10 ml
aliquot.
Spectrophotometric determination
Place the solution made up with pH 1.4 buffer in the reference beam and the solution made
up with pH 8 buffer in the sample beam. Read the optical density over the range 290-340 nm
{33,480-29,410 cm-1). The maximum absorption is at approximately 312 nm (32,050 cm-1)
and the E/rvalue at this wavelength is 144.
It is advisable to check the E % value and the wavelength for maximum absorption lcm
using a sample of pure hexachlorophane. Calculate the percentage hexachlorophane from the
observed reading.