Part 2

RAW MATERIAL SPECIFICATION
WATER
Items Specifications Methods
Appearance Match to standard 1476468
Odor Match to standard 2846466
Microbial Content Test

(Total Plate Count) <=100cfu/g 9184535
(Absence of Psuedomonas species) Must Pass 9413173
(Absence of E. Coli) Must Pass 9413173
ACACIA SENEGAL GUM EXTRACT
Microbial Content Test <=100 cfu/g 9184535

BUTYLENE GLYCOL
Identification Match to standard JSCI methods
Specific Gravity 1.004 – 1.007 2987147

SODIUM BENZOATE
Identification Match to standard JSCI method
Assay 99.0 % minimum JSCI method

METHYLPARABEN
Identification Match to standard USP method
Assay 99.0 – 100.5 % USP method
Melting Point 125 – 128 C USP method

PERFUME
Specific Gravity 0.94 - 0.98 2987147
Microbial Content Test N/A

(Total Plate Count)
CERTIFICATE
The perfume described below complies with the latest IFRA (The International Fragrance
Association) guidelines.
Perfume name / code : Rose ver 1

Name of Supplier : Max Fragrance Co. Ltd..
Address of supplier: 35894 Smiling Street, Sydney, NSW, Australia
________________________________________________ _____________
M. Jones Date
Senior Scientist
Research & Development
ASEAN Company
Part II. A Specifications and test methods of raw material / ingredients

- For fragrance materials, specify the name and code number of the fragrance,
name and address of the supplier, declaration of compliance with the latest IFRA
guidelines
SCCP/0873/05
EUROPEAN COMMISSION
HEALTH & CONSUMER PROTECTION DIRECTORATE-GENERAL
Directorate C - Public Health and Risk Assessment

C7 - Risk assessment
SCIENTIFIC COMMITTEE ON CONSUMER PRODUCTS
SCCP
Extended Opinion on
the Safety Evaluation of Parabens
Adopted by the SCCP

by written procedure on 28 January 2005
SCCP/0873/05
Opinion on the safety evaluation of parabens
TABLE OF CONTENTS
1. BACKGROUND 3
2. TERMS OF REFERENCE 3
3. OPINION 4
4. CONCLUSION 8
5. MINORITY OPINION 10
6. REFERENCES 10
7. ACKNOWLEDGEMENTS 11
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SCCP/0873/05
1. BACKGROUND
Parabens (4-Hydroxybenzoic acid, its salts and esters) are regulated by Cosmetic Directive
76/768/EEC, Annex VI, part 1, reference 12 and can therefore be used as a preservative up to a
maximum concentration of 0.4 % in the finished product for 1 ester and up to 0.8 % for mixtures
of esters. The substances are marketed with the symbol (+) and therefore may also be added to
cosmetic products in concentration other than those laid down in Annex VI for other purposes
apparent from the presentation of the product.
In its opinion of 17 February 1999 (SCCNFP/0125/99) concerning the restrictions on materials

listed in Annex VI of Directive 76/768/EEC on cosmetic products, the Scientific Committee on
Cosmetic Products and Non-Food Products intended for Consumers (SCCNFP) stated that those
substances indicated by (+) in Annex VI, when incorporated into cosmetic formulations for non-
preservative functions, should be subjected to the same restrictions in usage levels and warnings
as when used for preservative effects.
If a preservative marked with the symbol (+) is added for non-preservative purpose to a cosmetic
product in a concentration higher than that laid down in the Annex VI, data to substantiate its
safety should be submitted to the SCCNFP.
2. TERMS OF REFERENCE
The SCCP was asked to answer the following questions:
1. Do the data provided justify a concern that parabens when used up to the maximum
authorized concentration in cosmetic products might pose a risk to the consumer?
2. If yes, do the data provided justify a change in the maximum concentration for their use in
cosmetic products and would this concentration apply to all parabens used (methyl-, ethyl-,
propyl- and butylparaben)?
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SCCP/0873/05
3. OPINION
3.1. Recent Official Reports on Parabens
Parabens are the alkyl esters of p-hydroxybenzoic acid and are allowed as antimicrobial
preservatives for use in food products, medicinal products and cosmetics.
Recently, three important official reports have been issued on the toxicity profile and safety of
the use of parabens in consumer products. They were based upon a previous report of the
Scientific Committee on Food [SCF 1994].
3.1.1. The Scientific Committee on Food (SCF), SCF 1994
a) In a document issued 25 February 1994 on p-hydroxybenzoic acid alkyl esters and their
sodium salts, the SCF reported that acute toxicity of parabens was only seen at high dosages.
All the parabens produced similar symptoms with rapid onset of ataxia, paralysis and central
nervous system depression, resembling anaesthesia, suggesting their toxicity is related
mainly to the free acid. With non-lethal dosages, recovery is prompt.
b) In vitro and in vivo mutagenicity studies provided no evidence of genotoxicity for methyl,
propyl and butyl paraben. A carcinogenicity study with butyl paraben in the mouse was
reported to be negative, although the study appeared not being performed according to the
appropriate guidelines.
c) Reproduction and teratogenicity studies in the rat with ethyl paraben (up to 10% in the diet)
found no adverse effects on reproductive performance. Foetal anomalies, however, were
observed, though without a clear dose-response relationship. For this reason, a new oral
teratogenicity study in the rat was requested.
d) The absorption, metabolism and excretion of parabens had been studied in rats, rabbits, dogs
and humans. The methyl, ethyl and propyl esters of p-hydroxybenzoic acid appeared to be
well absorbed and the ester linkage was assumed to be readily hydrolyzed. Urinary excretion
of the unchanged esters was very low, usually less than 1% of the administered dose. Butyl
paraben was suspected to follow a different metabolite pathway, but studies in dogs had
shown no evidence of accumulation of either parent compound or metabolites in the tissues.
e) A number of special studies revealed that the parabens (in particular propyl and butyl
paraben) were able to induce cell proliferation in the forestomach and glandular stomach of
rats. A supplementary study in the rat on propyl paraben, given as a solution instead of a
ground powder, was requested.
f) Finally, the report stated several subchronic and chronic toxicity tests conducted in rats, dogs
and mice, resulting in a NOAEL-value of 1000 mg/kg bw/day. Based on this value, the EC
Scientific Committee on Food established a temporary Acceptable Daily Intake (ADI) of up
to 10 mg/kg as the sum of methyl, ethyl and propyl paraben and their sodium salts. That ADI
was temporary since the Committee asked for some additional information with regard to the
reproductive effects and more data on the cell proliferation effect of the compounds in the rat
forestomach.
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SCCP/0873/05
3.1.2. The European Food Safety Authority (EFSA), EFSA 2004
a) Newly available studies on the developmental toxicity of methyl paraben in rats, mice,
hamsters and rabbits were evaluated and no evidence of developmental toxicity up to
300 mg/kg bw/day (rabbits) or 550 mg/kg bw/day (rodents) was observed.
b) In the re-evaluation of the proliferative effects of parabens on forestomach cells in rats, it

was concluded that this effect only occurred above a certain threshold exposure that is not
reached in man.
c) An evaluation was also undertaken of the estrogenicity of parabens in vitro. However, for
methyl, ethyl and propyl paraben, such activity was not detectable in vivo using uterotrophic
assays (oral and s.c.) in mice and rats. On the contrary, for butyl and isobutyl paraben (not
used in food), a positive effect was seen after s.c. injection. For the major metabolite, p-
hydroxybenzoic acid, no effect was present.
d) Dietary administration of methyl and ethyl paraben to juvenile male rats had no effect on sex
hormones and reproductive organs at dosage levels up to 1000 mg/kg bw/day. Thus the
NOAEL for methyl and ethyl paraben is 1000 mg/kg bw/day.
For propyl paraben impaired spermatogenesis, reduced testosterone levels and reduced sperm
cell count numbers were observed and a LOAEL of 10 mg/kg bw/day was established.
e) The EFSA report came to the conclusion that the ADI of up to 10 mg/kg remains in place for
the sum of methyl and ethyl paraben and their sodium salts on the basis of a NOAEL of
1000 mg/kg.
For propyl paraben on the other hand, this ADI was not considered appropriate. An ADI for
the propyl ester could not be established because of the lack of a clear NOAEL.
3.1.3. The Danish Institute of Food and Veterinary Research, Anonymous 2004
In September 2004, the Danish Institute of Food and Veterinary Research issued a report entitled
"Note on Parabens in Food, Cosmetics and Consumer Products".
This document refers to the EFSA report for a full overview of the available toxicity data on
parabens, and focuses on the problems of endocrine disrupting potential, effects on the male
reproductive system, skin penetration of parabens, breast cancer & paraben-containing
cosmetics, possible interactions between different xenoestrogens, and possible low doses effects.
The authors repeat the conclusions of the EFSA report [EFSA 2004] and do not raise additional
concerns with regard to the use of parabens in cosmetics [Anonymous 2004].
3.1.4. The Norwegian Institute of Public Health (NIPH), Paulsen and Alexander 2003
In 2003, the NIPH published a report briefly summarizing the toxicity of the parabens and
studying in particular the alleged endocrine disrupting potential of the molecules. The authors
conclude that:
5
SCCP/0873/05
a) Different parabens have varying estrogenic potential in cell cultures and animal studies, but
their potency is 1000 to 1,000,000 times lower than the potency of 17β-estradiol or
testosterone.
b) In order to perform a revised and complete risk evaluation, data on reproduction in long-term
animal experiments, data from multiple generation experiments and more detailed
knowledge on the pharmacokinetics of parabens under use conditions, are required.
c) A preliminary risk assessment followed by the calculation of the Margin of Safety (MoS) for
the use of parabens as a preservative in cosmetic products, leads to worst case MoS's of 122
and 73 for adults and children, respectively.
In these calculations, the following parameters were considered:
- NOAEL (developmental toxicity of propyl and butyl paraben) 10 mg/kg bw/day

- total global exposure to all cosmetic products: 17.7 g
- percutaneous absorption (based on human in vitro studies): 3.5%
- contribution of dietary parabens: not considered (very small)
- mean human body weight: 60 kg
- maximum permitted concentration of paraben mixture: 0.8%
- larger body surface per body mass of children versus that of adults: factor 1.7
- extensive biotransformation of parabens into
p-hydroxybenzoic acid (liver, skin): not accounted for
d) Neither interactions, additive or synergistic effects, nor effects at doses below the ones
tested, are likely.
3.2. Toxicity Profile of the parabens used in cosmetic products
Viewing the availability of the four above-mentioned official reports, the toxicity profile of the
parabens will only be briefly summarised in the current report and more detailed sections will be
dedicated to the specific potential problem areas.
3.2.1. General toxicology
Based on acute, subacute and chronic toxicity studies in rats, dogs and mice, parabens have been
proven to be practically non-toxic, not carcinogenic, not genotoxic nor co-carcinogenic, and not
teratogenic. A NOAEL value of 1000 mg/kg bw/day was accepted for all esters.
Parabens are not expected to accumulate in tissues and the ester linkage of the parabens is
expected to be readily hydrolyzed [SCF 1994].
3.2.2. Estrogenic effects of parabens
In a number of in vitro studies, such as the recombinant yeast estrogen screen, parabens have
proven being able to bind the estrogen receptor, to activate genes controlled by these receptors,
and to stimulate cell growth and increase the level of immune reactive estrogen receptor protein.
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SCCP/0873/05
The estrogenic potency increases with increasing length and branching of the alkyl side chains
(methyl < ethyl < propyl < butyl < isobutyl), but remains at all times 1000 to 1,000,000 times
below the potency of 17β-estradiol. p-Hydroxybenzoic acid, the common metabolite of all
parabens, appeared to be inactive in the in vitro assays.
The in vivo estrogenic activities of parabens have been tested in uterotrophic assays employing
either immature female rodents or adult ovariectomized female rodents after oral, subcutaneous
or dermal administration. Again, butyl paraben appeared being more potent than propyl, ethyl
and methyl paraben, and again the values remained several magnitudes of order below the
potency of 17β-estradiol. Conflicting results have been reported for p-hydroxybenzoic acid
tested in vivo. One study claims that it has no estrogenic effect; another study gives potency
values 1000-fold below the 17β-estradiol level [EFSA 2004, Anonymous 2004, Paulsen and
Alexander 2003].
3.2.3. Effects of parabens on the male reproductive system
Methyl, ethyl, propyl and butyl paraben have been examined for effects on the reproductive
organs in the male offspring of rats and mice.
Male neonatal Wistar rats s.c. injected with butyl paraben at 2 mg/kg bw/day on postnatal
days 2 to 18 showed no detectable effects on any reproductive parameter [Fisher et al. 1999].
This study, however, demonstrated only very minor effects for compounds such as genistein,
bisphenol A and octylphenol administered at high dosages.
On the contrary, 10 mg/kg bw/day administered to post-weaning male Wistar rats for eight
weeks through their diet caused a decrease of the cauda epididymal sperm reserve, a decrease in
sperm count, in daily sperm production and in serum testosterone [Oishi 2001].
In another study, pregnant Sprague Dawley rats were s.c. injected with daily dosages of 100 or
200 mg butyl paraben/kg bw from gestational day 6 to postnatal day 20, and the offspring were
examined. Both tested dosages showed clear effects, amongst which a decrease in sperm count
and sperm motile activity in the epididymus [Kang et al. 2002].
Administered to ICR (Cjr:CD-1) mice in the diet for 10 weeks at dosage levels of 14.4, 146 and
1504 mg/kg bw/day, butyl paraben showed some clear effects at the two highest dosage levels,
including increased epididymal weights, despite of a decrease in testis spermatid count and in
serum testosterone levels. The authors were unable to explain this discrepancy [Oishi 2002a].
In 2002, Darbre et al. published a study on the estrogenic activity of isobutyl paraben in vitro
and in vivo (subcutaneous administration). The authors conclude that the studies clearly show
that isobutyl paraben is more potent, but the doses used in the in vivo test were not higher than
the ones tested for all other parabens (1.2 and 12 mg isobutyl paraben/mouse, equivalent to
approximately 24 and 240 mg/kg bw). [Darbre et al. 2002].
As far as propyl paraben is concerned, a four-week administration via the diet (0, 10, 100,
1000 mg/kg bw/day) to the Wistar rat showed similar effects as caused by butyl paraben, though
at a dosage of 100 mg/kg bw/day. The propyl ester caused only minor effects at
10 mg/kg bw/day. No dose-response relationship was observed [Oishi 2002b].
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SCCP/0873/05
Recently, methyl and ethyl paraben have been tested for effects on secretion of sex hormones
and the Wistar rat male reproductive system. Dosages of 103 and 1030 mg/kg bw/day failed to
induce adverse effects [Oishi 2004].
Taking together the above-mentioned studies, it is clear that butyl paraben shows the highest
potency with regard to effects to the male reproductive system. This is in accordance with the
results of the estrogenicity tests. It was also concluded that, in order to define an exact NOAEL
for propyl paraben, additional studies were necessary [EFSA 2004, Anonymous 2004].
Very recently, a detailed developmental toxicity evaluation of butyl paraben given by oral
gavage to Sprague-Dawley rats became available [Daston 2004]. Dosage levels of 0, 10, 100 and
1000 mg/kg bw/day were administered on gestation days 6-19 (sperm positive day being
gestation day 0). Foetuses were evaluated on gestation day 20. The highest dosage level caused
decreases in maternal weight gain (significant during gestation day 18-20 interval). Maternal
food consumption was, however, also decreased in that group for the gestation days 6-20
interval. No differences from the control group could be observed in any of the developmental
parameters including embryo/foetal viability, foetal weight, malformations or variations. The
maternal NOAEL for butyl paraben was established on 100 mg/kg bw/day. It was further
concluded that butyl paraben does not have the potential to cause developmental toxicity in the
Sprague-Dawley rat at oral dosages up to 1000 mg/kg bw/day [Daston 2004].
3.2.4. Breast cancer and the use of paraben-containing cosmetics
The suggested link between the use of (paraben-containing) underarm cosmetics and breast
cancer has been discussed and refuted in a separate SCCP opinion on Parabens, underarm
cosmetics and breast cancer [SCCP/0874/05]. This issue will therefore not be reconsidered in the
present opinion.
4. CONCLUSION
• Is the concern justified that parabens, when used up to the maximum authorized
concentration in cosmetic products, might pose a risk to the consumer?
Methyl and ethyl paraben
Previous toxicological data have led the EU Scientific Committee on Food to establish an
Acceptable Daily Intake (ADI) level of 10 mg/kg bw/day as the sum of methyl, ethyl and propyl
p-hydroxybenzoic acid esters and their sodium salts [SCF 1994].
With the most recent findings on the alleged estrogenic effects of the parabens and more
importantly with the newly discovered effects on the male reproductive system, as described
under point 3.3 of this opinion, the European Food Safety Authority reviewed the safe use of
parabens in food. The Panel on Food Additives, Flavourings, Processing Aids and Materials in
Contact with Food came to the conclusion that the ADI remained unchanged for methyl, ethyl
paraben and their sodium salts, but that for propyl paraben no ADI could be established, by lack
of a clear NOAEL value.
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SCCP/0873/05
Therefore, it is the opinion of the SCCP that methyl and ethyl paraben can be safely used up to
the maximum authorized concentration as actually established (0.4%).
Propyl, butyl and isobutyl paraben
Propyl paraben
- The developmental rat study provided for propyl paraben failed to indicate a clear NOAEL
value, but it suggested that the potency of propyl paraben is clearly lower than the potency
of butyl paraben [Oishi 2002b]. For the latter, a NOAEL value of 2 mg/kg bw/day was
proposed. This value was obtained in a study where the compound was orally administered
to male Wistar rats for 17 days and the investigated reproductive parameters included
testicular weight, aquaporin-1 immunoexpression, rete testis distention and efferent duct
epithelial cell height [Fisher 1999].
- The LOAEL-value for propyl paraben in Wistar rats was shown to be 10 mg/kg bw/day
[Oishi 2002a].
- The major adverse effects of concern caused by propyl paraben involve the male
reproductive system. It must not be forgotten, however, that the majority of cosmetic
products will be used by females. Especially the cumulative cosmetic exposure value of
17.79 g/day is a clear overestimation for the normal male population.
Butyl and isobutyl paraben
- For butyl paraben a NOAEL value in Cjr:CD-1 mice (oral study) of 2 mg/kg bw/day has
been proposed [Oishi 2002b].
- Very recently, a developmental study on butyl paraben became available. In that study, it
was shown that the maternal NOAEL of butyl paraben was 100 mg/kg bw/day and that the
compound did not have the potential to cause developmental toxicity in the Sprague-
Dawley rat.
It is known that potent estrogens interfere with the development of the reproductive
system, placental function, embryonic growth, embryo viability and maintenance of
pregnancy [Daston et al. 1997]. Although indirectly, these parameters could be responsive
indicators of estrogenicity. This leads to the suggestion that butyl paraben did not have a
strong estrogenic potential during the developmental study.
It is the opinion of the SCCP that the available data do not enable a decisive response to the
question of whether propyl, butyl and isobutyl paraben can be safely used in cosmetic products
at individual concentrations up to 0.4%.
Therefore industry is requested to provide the complete developmental toxicity dossier of these
three esters of p-hydroxybenzoic acid. The reason for this request is that the current opinion is
based upon extended literature data and there is a significant possibility that additional in vivo
data have been generated within industry without having been submitted to the SCCP.
9
SCCP/0873/05
• If yes, do the available data justify a change in the maximum concentration for their use in
cosmetic products and would this concentration apply to all parabens used (methyl-,
ethyl-, propyl- and butylparaben)?
Methyl and ethyl paraben
For the methyl and ethyl p-hydroxybenzoic acid esters, the maximum authorized concentrations
remain unchanged.
Propyl, isopropyl, butyl and isobutyl paraben
As the present discussion is based solely upon data in the literature, it is the SCCP's opinion that
more information is needed in order to formulate a final statement on the maximum
concentration of propyl, isopropyl, butyl and isobutyl paraben allowed in cosmetic products.
More specifically, the following data are requested before end of March 2005:
- full descriptions of available in vitro percutaneous absorption studies;

- a complete dossier with regard to the reproductive and developmental toxicity of propyl,
isopropyl, butyl and isobutyl paraben, with special focus on the male reproductive system.
5. MINORITY OPINION
Not applicable
6. REFERENCES
Anonymous, Note on Parabens in Food, Cosmetics and Consumer Products. Danish Institute of
Food and Veterinary Research, Department of Toxicology and Risk Assessment, September
2004.
Darbre P.D., Byford J.R., Shaw L.E., Horton R.A., Pope G.S., Sauer M.J., Oestrogenic activity
of isobutylparaben in vitro and in vivo. J Appl Toxicol. 22(4), 219-26 (2002).
Daston G.P., Gooch J.W., Breslin W.J., Shuey D.L., Nikiforov A.I., Fico T.A., Gorsuch J.W.
Environmental estrogens and reproductive health: a discussion of the human and environmental
data. Reprod Toxicol. 11(4), 465-481 (1997).
Daston G.P. Developmental toxicity evaluation of butylparaben in Sprague-Dawley rats. Birth
Defects Res Part B Dev Reprod Toxicol. 71(4), 296-302 (2004).
EFSA, Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and
Materials in Contact with Food on a Request from the Commission related to para
hydroxybenzoates (E214-219), Question number EFAS-Q-2004-063. adopted on 13 July 2004
The EFSA Journal 83, 1-26 (2004).
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SCCP/0873/05
Fisher J.S., Turner K.J., Brown D., Sharpe R.M. Effect of neonatal exposure to estrogenic
compounds on development of the excurrent ducts of the rat testis through puberty to adulthood
Environ Health Perspect. 107(5), 397-405 (1999).
Kang KS, Che JH, Ryu DY, Kim TW, Li GX, Lee YS. Decreased sperm number and motile
activity on the F1 offspring maternally exposed to butyl p-hydroxybenzoic acid (butyl paraben).
J Vet Med Sci. 64(3), 227-235 (2002).
Oishi S. Effects of butylparaben on the male reproductive system in rats. Toxicol Ind Health.
17(1), 31-9 (2001).
Oishi S. Effects of propyl paraben on the male reproductive system. Food Chem Toxicol.
40(12), 1807-1813 (2002a).
Oishi S. Effects of butyl paraben on the male reproductive system in mice. Arch Toxicol. 76(7),
423-429 (2002b).
Oishi S. Lack of spermatotoxic effects of methyl and ethyl esters of p-hydroxybenzoic acid in
rats. Food Chem Toxicol. 42(11), 1845-1849 (2004).
Paulsen J.E. and Alexander J. Evaluation of parabens in cosmetic products. Norwegian Institute
of Public Health (2003).
SCCP/0874/05 - Opinion of the Scientific Committee on Consumer Products on Parabens,
Underarm Cosmetics and Breast Cancer, adopted by written procedure on 28 January 2005
SCF. Opinion on p-hydroxybenzoic acid alkyl esters and their sodium salts expressed on 25
February 1994. European Commission, Reports of the Scientific Committee for Food (Thirty-
fifth series), http://europa.eu.int/comm/food/fs/sc/scf/reports/scf_reports_35.pdf (consulted Nov
2004), p.9-12.
7. ACKNOWLEDGEMENTS
Members of the working group are acknowledged for their valuable contribution to this opinion.
The members of the working group are:
Dr. C. Chambers Prof. J.-P. Marty

Prof. R. Dubakiene Dr. S.C. Rastogi
Dr. R. Grimalt Prof. J. Revuz
Dr. B. Jazwiec-Kanyion Prof. V. Rogiers (Rapporteur)
Prof. V. Kapoulas Prof. T. Sanner
Prof. J. Krutmann Prof. G. Speit
Prof. C. Lidén Dr. I.R. White (Chairman)
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SCCP/0891/05
EUROPEAN COMMISSION
HEALTH & CONSUMER PROTECTION DIRECTORATE-GENERAL
Directorate C - Public Health and Risk Assessment

C7 - Risk assessment
SCIENTIFIC COMMITTEE ON CONSUMER PRODUCTS
SCCP
OPINION ON
Benzoic Acid and Sodium Benzoate
Adopted by the SCCP

during the 4th plenary of 21 June 2005
SCCP/0891/05
Opinion on Benzoic Acid and Sodium benzoate
TABLE OF CONTENTS
1. BACKGROUND ………………………………………………… 3
2. OPINION ………………………………………………… 4
3. CONCLUSION ………………………………………………… 25
4. MINORITY OPINION ………………………………………………… 25
5. REFERENCES ………………………………………………… 25
6. ACKNOWLEDGEMENTS ............................................................................ 29
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SCCP/0891/05
1. BACKGROUND
1.1. Background
The Scientific Committee on Cosmetic Products and Non-Food Products intended for
Consumers (SCCNFP) was requested to review the data submitted to support the safety of
benzoic acid, its salts and esters (COLIPA1 No. P 2), when used at concentrations other than
those laid down in Annex VI to Directive 76/768/EEC as preservatives, for other specific non-
preservative purposes apparent from the presentation of the products.
The SCCNFP adopted an opinion on 4 June 2002 on Benzoic acid and Sodium benzoate
(SCCNFP/0532/01, final). In its opinion the SCCNFP stated, that “the SCCNFP can only assess
the safety of substances for which appropriate data has been submitted for evaluation. Safety
assessment is specific and not generic. Only toxicological data for benzoic acid and its salt
sodium benzoate have been made available for review. Therefore, there is no review of other
salts of benzoic acid or any of its esters. These will require separate evaluation when the
necessary data have been made available.”
It was stated furthermore, that “the SCCNFP does not find the submission appropriate for a
safety evaluation of benzoic acid and sodium benzoate for the applied “other uses” in cosmetic
products”.
Recently, the European Commission received Submission II on Benzoic acid, its salt and esters.
2. TERMS OF REFERENCE
1. On the basis of provided data the SCCP is asked to assess the risk to consumers when
Benzoic acid, its salts and esters are used for non-preservative purposes in cosmetic rinse-off
products at a maximum concentration of 2.5 % and in cosmetic oral-care products at a
maximum concentration of 1.7 %, and in leave-on products up to 0.5%.
2. Does the SCCP recommend any further restrictions with regard to the use of Benzoic acid,
its salts and esters safe when used for non-preservative purposes in cosmetic products?
1
COLIPA – European Cosmetics Toiletry and Perfumery Association
3
SCCP/0891/05
3. OPINION
3.1. Chemical and Physical Specifications
3.1.1. Chemical identity
3.1.1.1. Primary name and/or INCI name
Benzoic acid (INCI name)

Sodium benzoate (INCI name)
3.1.1.2. Chemical names
Benzoic acid Benzene carboxylic acid; benzene formic acid; carboxybenzene; benzene
carboxylic acid; phenylcarboxyl acid; phenylformic acid, E210
Sodium benzoate E211
3.1.1.3. Trade names and abbreviations
3.1.1.4. CAS / EINECS / ELINECS number
CAS EINECS
Benzoic acid : 65-85-0 200-618-2
Sodium benzoate : 532-32-1 208-534-8
3.1.1.5. Structural formula
Benzoic acid Sodium benzoate
3.1.1.6. Empirical formula
Benzoic acid : C7H6O2

Sodium benzoate : C7H5O2Na
3.1.2. Physical form
Benzoic acid : solid

Sodium benzoate : solid
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SCCP/0891/05
3.1.3. Molecular weight
Benzoic acid : 122.13

Sodium benzoate : 144.11
3.1.4. Purity, composition and substance codes
No data
3.1.5. Impurities / accompanying contaminants
No data
3.1.6. Solubility
Benzoic acid : 2.91 g/l in water at 20oC

Sodium benzoate : 556 g/l in water at 20oC, hygroscopic
3.1.7. Partition coefficient (Log Pow)
Benzoic acid : 1.88

Sodium benzoate : -2.269
3.1.8. Additional physical and chemical specifications
Benzoic acid
Organoleptic properties : /
Melting point : 122 °C
Boiling point : 249.2°C
Flash point : /
Vapour pressure : 0.0011hPa
Density : 1.321 g/cm3 at 20°C
Viscosity : /
pKa : -2.269
Refractive index : /
Stability : /
Sodium benzoate
Organoleptic properties :
Melting point : >300.0 °C
Boiling point : 464.9°C
Flash point : /
Vapour pressure : 0.0011hPa
Density :
Viscosity : /
pKa : -2.269
Refractive index : /
Stability : /
5
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3.2. Function and uses
Benzoic acid is a natural ingredient occurring in many foodstuffs and in plant extracts.
Benzoic acid, its salts and esters are used as preservatives in cosmetic products, with a maximum
concentration of 0.5 %, (calculated as acid), as regulated by the EU Cosmetics directives
76/768/EEC.
This request is for use for non-preservative purposes in cosmetic rinse-off products, at a
maximum concentration of 2.5 %, and in cosmetic oral-care products, at a maximum
concentration of 1.7 %, and in leave-on products, up to 0.5%. However these purposes are not
specified.
Other uses for benzoic acid and its salts include regulated use as food preservatives, most
suitable for foods, fruit juices, and soft drinks in an acidic pH range. In the EU, there are
regulations controlling the maximum levels of benzoic acid and its salts for use in foodstuffs
ready for consumption and the specific purity criteria of food additives. The levels are expressed
as the free acid.
Non alcoholic drinks 150 mg/l

Alcoholic drinks 200 mg/l
Jams & jellies 500 mg/kg
Aspic 500 mg/kg
Ref.: AR 1, AR 2
In the United States, benzoic acid and sodium benzoate are on the FDA list of substances that are
generally recognized as safe (GRAS). Both may be used as antimicrobial agents, flavouring
agents and as adjuvants with a current maximum level of 0.1% in food. The FDA has not
determined whether significantly different conditions of use would be GRAS. The FDA has
sought fully up-to-date toxicology information.
Ref.: AR 3
Benzoic acid is used in oral medicines up to 0.15%, in parenteral medicines up to 0.17% and in
topical drugs up to 0.2%. Benzoic acid is used as an active ingredient in anti-fungal cream with
salicylic acid (3.0%) up to 6%.
Sodium Benzoate, expressed as benzoic acid, is permitted in oral medicines up to 0.5%, in
parenterally administered up to 0.5%.
Ref.: AR 4, AR 5
Benzoic acid is also an intermediate in the synthesis of phenol and caprolactam. Other end
products include sodium and other benzoates, benzoyl chloride, and diethylene and dipropylene
glycol dibenzoate plasticizers. Sodium benzoate is primarily a preservative and corrosion
inhibitor (e.g., in technical systems as an additive to automotive engine antifreeze coolants).
6
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3.3. Toxicological Evaluation
3.3.1. Acute toxicity
3.3.1.1. Acute oral toxicity
Acute oral toxicity information is based on summary information, no dossiers were provided.
The indication is that the substances can be considered to possess low acute oral toxicity. The
calculated LD50 values taken from the available experiments on the acute oral toxicity of
Benzoic Acid and Sodium Benzoate are listed in the following tables:
Benzoic Acid
Species LD50 [mg/kg] Reference
Rats 1700
2530 4
Mice 1940 5
2370 6
Sodium Benzoate
Species LD50 [mg/kg] Reference
Rats 1714 7
2100 fasting 8
3140 9
3450 not fasted 8
4070 10
3.3.1.2. Acute dermal toxicity
Benzoic acid
Rabbit LD50 > 5 000 mg/kg
LD50 > 10 000 mg/kg
Ref.: 14, citation only
3.3.1.3. Acute inhalation toxicity
No data submitted with Submission II (2004)
Benzoic acid
Rats LC50 > 0.026 mg/l/h
Ref.: 14, citation only
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3.3.2 Irritation and corrosivity
3.3.2.1. Skin irritation
Animal studies
Benzoic Acid
Guideline : OECD 404 (1981), EEC test method B.4, Annex V of EEC Directive
84/449/EEC (September 1984)
Species/strain : New Zealand albino rabbit
Group size : 3 females
Test substance : 0.5 g of benzoic acid moistened with 0.25 ml Milli-RO water
Batch : /
Purity : 99.5%
GLP : in compliance
A paste, (0.5 g benzoic acid and 0.25 ml water), was applied evenly to 6 cm2 Metalline on a
permeable tape (Micropore). This was applied to the right flank of the rabbit. A control patch
without the test substance was applied to the left flank. These were left in place for 4 hours. The
test site was cleaned first with a dry tissue and then by swabbed with a dampened tissue. The
skin was examined for erythema, eschar formation and oedema at 1, 24, 48 and 72 hours after
removal of the patches.
Results
Two animals showed slight erythema initially. 1 of these also showed slight oedema up to 24 h.
This was resolved by 48 h. There was no indication of a systemic effect.
The results of this study indicate that the test item, benzoic acid, was minimally irritating
(modified primary irritation index (PII) of 0.5) when applied to the intact rabbit skin under semi-
occlusive patch conditions. The test substance does not need to be labelled as a skin irritant.
Ref.: 12
In submission II, three additional animal tests confirmed the low irritation potential of Benzoic
Acid were summarized in a table. These were also provided in Submission I, but are only
citations or summary information.
Species Exposure Skin irritation Reference

rabbits neat Benzoic acid/occluded / 24 h mild 13
rabbits 500 mg dry powder/ 24 and 72h PII = 1.66/8.00 14
rabbits 500 mg dry powder/ semiocclusive / 24 h not irritating 15
Sodium Benzoate
Guideline : OECD 404 (1981)
Test substance : 0.5 g sodium benzoate moistened with 0.25 ml Milli-RO water
Batch : /
Purity : > 99.5%
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GLP : in compliance
Approximately 24 hours prior to the treatment, the dorsal fur was shaved. A paste, (0.5 g sodium
benzoate and 0.25 ml water), was applied evenly to 6 cm² Metalline on a permeable tape
(Micropore). The 6 cm² patch was removed four hours after semi-occlusive contact. The skin
reactions were assessed approx. 1 hour, 24, 48 and 72 hours after termination of the exposure.
Under the conditions of the study, sodium benzoate was neither corrosive nor irritating (PII 0)
when applied to the intact rabbit skin under semi-occlusive patch conditions for four hours. The
test substance does not need to be labelled as a skin irritant. The animals did not show any
symptoms of systemic intoxication.
Ref.: 16
Application of 500 mg sodium benzoate as a dry powder to rabbit for 24 h; responses were
scored at end of treatment and after 48 h. It was considered not irritating.
There are no details of study.
Ref.: 17
Human Studies
Benzoic Acid
The results, presented only in scientific papers, are from studies both on volunteers and patients
from dermatological clinics. 2% benzoic acid in petrolatum over 46h did not irritate intact skin
of healthy volunteers. 24h application of 30% benzoic acid in ethanol was found to be the lowest
irritating concentration.
Ref. : Gad et al, 1986, Kligman, 1977, Frosch & Kligman, 1976 cited in 13
Chamber test (72 h/occlusive): 0.1 ml of 7.5 % and 15 % benzoic acid in ethanol on scarified
skin, 30 % benzoic acid in ethanol on normal skin (6 volunteers).
Results
Scarified skin : 7.5 % in ethanol, moderate irritant
15.0 % in ethanol : marked irritant with erosions
Normal skin : 30 % in ethanol, lowest irritant concentration
Ref.: 19, 21
Chamber test (20 min/occlusive), open test (30 min): 15 µl of 5 % benzoic acid in petrolatum, 15
atopic and 16 non-atopic patients. The atopics showed redness in both the chamber test, (73 %)
and the open test, (80 %). Non-atopics showed 80% redness in both the chamber test and in the
open test. There was no statistical difference between atopics and non-atopics.
Ref.: 20
8 out of 627 patients (1.3%) from dermatological clinics showed positive reactions to 5 %
benzoic acid, in petrolatum under an occlusive dressing for 24 or 48 h. At this concentration, the
authors suggest that these results could be interpreted as marginally irritating, rather than
allergic.
Ref.: 18
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Non-immunologic contact urticaria
Benzoic acid (5.0 % in petrolatum) was tested in an open test on 29 atopic and 74 non-atopic
persons.
Results
Contact urticarial reactions to benzoic acid were seen in 27/29 (93 %) of the atopics and in 64/74
(87 %) of the non-atopics.
In the chamber test, 20 min occlusion (recorded 10 min later) 0.1 % benzoic acid in petrolatum
and 0.05 % benzoic acid in water elicited reactions. When water was the vehicle, the reactions
were oedematous.
Ref.: AR6
Urticaria occurred in 1/10 and 5/12 healthy volunteers with 0.1 and 1.0% benzoic acid, following
20 min covered contact. It was not clear if this was an irritant or immune response.
Ref.: Clemmensen &Hjorth, 1982 cited in 13
Non-immune immediate contact reactions (NIICR), erythema and oedema, have been produced
in 78/80 women tested with 2% benzoic acid petrolatum but there was no correlation between
the susceptibility to NIICR and age, atopic status or tanning ability. In another study, it was
demonstrated that benzoic acid induced non-immune immediate contact reactions in the majority
of 200 volunteers with no specific skin condition. Approximately 10% of the volunteers
appeared particularly sensitive, reacting fairly strongly. The response, erythema or oedema, was
specific to an individual with no significant correlation with age or sex on the degree of NIICR.
Ref.: 54
The Concise International Chemical Assessment Document (CICAD) No. 26 conclusion on

benzoic acid and sodium benzoate was: “However, both substances are known to cause non-
immunologic immediate contact reactions. This effect is scarce in healthy subjects, while in
patients with frequent urticaria or asthma, symptoms or exacerbation of the symptoms were
observed”.
Ref.: 55
3.3.2.2. Mucous membrane irritation
Guideline : EEC test method B.5, Annex V of EEC Directive 84/449/EEC

Test substance : Benzoic acid
Purity : > 99.5%
Batch no : /
Dose : 77 mg as a fine powder
GLP : in compliance
The test substance was instilled in a single application. The product was poured into the right
conjunctival sac. After application, the lids of the treated eye were held closed for approximately
two seconds. The untreated left eye was used as a control. The degree of eye irritation was
10
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evaluated at immediately after dosing, 1, 24, 48 and 72 hours and 7, 14 and 21 days after
treatment.
After 60 min, Animal 1 showed no reaction to light, with translucent corneal opacity and iridial
injection. The other 2 also had slight corneal opacity. All 3 had moderate chemosis and slight
conjunctival redness.
The corneal opacity noted in the first animal increased to nacreous areas. This persisted for 72 h.
The translucent areas of opacity persisted up to Day 21. In Animal 2, the slight corneal opacity
also persisted up to Day 21, but was resolved in Animal 3 by Day 7. By Day 14, in Animal 1, the
iridial injection was resolved but no reaction to light. Animal 2 showed iridial injection on Day
2. The slight conjunctival redness in all animals increased to severe with a white/grey
discolouration. It persisted in Animals 1 and 2 up to Day 21 and in Animal 3 to Day 7. The
chemosis decreased slowly. In Animals 1 and 2, it was not completely resolved by Day 21 but it
was resolved in Animal 3 by Day 14. The animals did not show any symptoms of systemic
intoxication.
The estimated Draize score of 35 (60 min) is classed as severely irritating according to Kay and
Calandra. Under the EU criteria, it should be labelled as an eye irritant.
Comment
This experiment was conducted in 1988, but the information was not provided in Submission 1.
Ref.: 22
Guideline : OECD 405

Group size : 3 males
Test substance : Benzoic acid DAB 8
Batch no : /
Dose : /
GLP : /
treatment.
The authors considered benzoic acid was considered as a mild irritant, but the data provided is
inadequate.
Ref.: 23, cited in BUA (14)
Instillation of 50 mg benzoic acid into the conjunctival sac of 2 rabbits.
Results
Moderate irritation. This is just a summary.
Ref.: 15, cited in BUA (14)
Additional information that was not mentioned but quoted in the BUA reference, was a citation
of a single application of 100 mg dry powder to the rabbit eye. The irritation score was 65.0/110.
It was scored at 24, 48, and 72 h.
11
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Ref.: cited in BUA (14)
Comment
The data provided is inadequate.
Sodium benzoate
Guideline : OECD 405

Test substance : Sodium Benzoate
Purity : ≥ 99.5%
Batch no : /
Dose : 60 mg ground as a fine powder
GLP : in compliance
treatment.
Under the conditions of this study, sodium benzoate produced irritation of the conjunctiva that
was reversed within 14 days. There was no effect on the cornea or iris. It was considered mildly
irritating (Kay and Calendra score 9.3) and in the EC classification.
Ref.: 24
Application of 50 mg sodium benzoate/rabbit for 24 h; responses were scored at 24 h, 48 h and

72 h; post-exposure observation time: 7 d.
Results: not irritating.
Ref.: 17
3.3.3. Skin sensitisation
Benzoic acid
Animal tests
The results of two in vivo guinea pig assays, a Magnusson-Kligman test and a Buehler test were
reported. These were in-house tests with no further details. Benzoic Acid did not induce
sensitization in either test and was thus considered to have low skin sensitization potential.
Ref.: 25
12
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Human volunteer studies
Benzoic Acid at 2 % and 5 % in petrolatum did not induce sensitization in two maximization
tests.
25 human volunteers were given five 48 h patch tests (over a 10 d period) with 2 % benzoic acid
in petrolatum. None gave positive reactions when challenged 10-14 d after the induction phase
with a final 48 h closed patch test with 2 % benzoic acid in petrolatum.
This information is as a summary only.
Ref.: 26 and cited in BUA (14)
10 persons allergic to benzoyl peroxide were tested by a 48 h patch test with 5 % benzoic acid in
a hydrophilic petrolatum. No reactions at 48, 72 and 96 h.
Ref.: 27
In a cosmetic intolerance assay, 5202 patients with suspected allergic contact dermatitis were
patch tested (537 of the patients had a history of “intolerance” or allergy). Patch test conditions
were not specified, but allergic reactions was noted in 34 patients (0.7% incidence) to benzoic
acid.
Ref.: 28
The OECD SIDS report found both benzoic acid and sodium benzoate were non-sensitizing in
animal test but showed a very low incidence in humans (patients) tested by the patch test.
This report included 2 additional human patch test references that were not in the submissions.
The results showed that benzoic acid was an occasional or positive sensitizer (Rademaker &
Forsyth, 1989; Forsbeck & Skog, 1977)
Ref.: 56
Sodium benzoate
Human
There were 2 additional human patch test references that were not included with any of the
submissions. The first was a large cohort of 2045 patients, 5 gave positive results. These gave
occasional positive results (Brasch, J. et al., 1993). In the second test, patch tests gave positive
nonimmunologic contact urticaria (Nethercott, J.R., 1984).
Ref.: 56
3.3.4. Dermal / percutaneous absorption

14
C-labelled benzoic acid (4 - 40 µg/cm2 dissolved in acetone) was applied to excised human
skin in a static diffusion chamber. Samples of penetrated amounts were measured in 1- 2 hr
interval over the first 24 hrs and 3-6 hrs interval over the remaining days. No occlusion or
washing were applied and the investigation was determined at least in duplicate. A median of
45% of the applied benzoic acid was found in the receptor phase 48 hrs after application.
Ref.: 40
Percutaneous absorption of benzoic acid was studied in the Mexican hairless dog and in man.
14
C-labelled benzoic acid was injected subcutaneously and applied dermally to the neck skin of
dogs. Human data were obtained from earlier investigations. Excretion of benzoic acid in man
13
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was rapid, almost complete by day 3. In the dog, excretion was less extensive and greatly
prolonged. This was accounted for by the persistence of benzoic acid in the skin.
Maximum absorption rate in man was 3.0 %/h, total absorption was 42.6 % of applied dose.
This in vivo result is comparable with in vitro test results obtained. The total penetration of
radio-labelled benzoic acid was found to be approximately 43 % in a 5-day period. In this human
study, 6 test subjects received a dose of 4 µg/cm² Benzoic Acid dissolved in acetone. They were
allowed to wash their skin 24 hours after application of Benzoic Acid. It should be noted that the
exposure of the in vitro and in vivo studies were extremely long (2 days and 5 days,
respectively).
Ref.: 41
3.3.5. Repeated dose toxicity
3.3.5.1. Repeated Dose (28 days) oral / dermal / inhalation toxicity
Oral
Benzoic Acid and sodium benzoate show a low toxic potential in rodents in repeat dose studies.
Benzoic Acid
Guideline : Directive 84/449/EEC, B.1 "Acute toxicity (oral)”
Species/strain : 50 Spartan rat
Group size : 5 male/ female
Test substance : technical grade benzoic acid
Purity : /
Batch no : /
Dose : 500, 1250, 1984, 3150, and 5000 mg/kg.
Vehicle : corn oil
GLP : /
The test compound was suspended in corn oil and administered orally. Volumes of 10 ml/kg bw
were administered at all dosage levels.
All surviving rats, males and females, exhibited normal body weight gains during the 14 day
observation period. The acute oral LD50 of benzoic acid in male albino rats was calculated to be
2742 mg/kg (2279-3299 mg/kg).
The acute oral LD50 of benzoic acid in female albino rats was calculated to be 2360 mg/kg
(2042-2726 mg/kg).
A combined acute oral LD50 for benzoic acid in male and female albino rats was calculated to
be 2565 mg/kg (2292-2870 mg/kg).
Ref : IUCLID, Unpublished study (IRDC#163-282). Acute Toxicity Studies in Rats and Rabbits.
(1974)
Comment: This study is available on IUCLID but was not submitted.
In rats, 648 mg/kg bw/d in feed for 28 days, has no effects.

Ref.: 14
14
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Sodium benzoate
0.5, 1, 2, 4, and 8 % sodium benzoate in drinking water were administered for 35 days to groups
of four female and four male Swiss albino mice.
In the 8 % dose level (approx. 24 g/kg bw/d) all mice died within 3 weeks. In the 4 % dose level
(approx. 12 g/kg bw/d) 3 male and 3 female mice died within the 35-day observation period. The
bodyweight of the surviving mice was substantially reduced. The 2 % dose level was chosen for
a chronic toxicity (carcinogenicity) study.
Ref.: 42
In another study the body weight was slightly decreased at 2200 mg/kg bw/d. Groups of 6 male
and 6 female Sherman rats were given 2 % (approx. 2.0 to 2.4 g/kg bw) or 5 % (5.7 g/kg bw for
females and 7.8 g/kg bw for males) sodium benzoate in the diet for 28 days. In the 5 % dose
group, all female rats died by day 11 and males by day 13. In the 2 % dose group a slight
significant body weight depression was observed in male rats.
Ref.: 43
Dose levels from 16 to 1090 mg sodium benzoate/kg bw were given to groups of 10 rats (5 male
and 5 female) for 30 days with the diet. No dose related adverse effects were observed.
Ref.: 10
Inhalation
Guideline : similar to OECD guideline 403
Species/strain : Rat, Sprague-Dawley (Charles River CD).
Group size : 10 males + 10 females per dose
Test substance : benzoic acid
Batch no : 59230350
Purity : /
Dose : 0, 0.025, 0.25 and 1.2 mg/l
Exposure : dust aerosol exposure, (diameter 4.7 µ) 6 h/day for 5 days/week
GLP : in compliance
At 1.2 mg/l two animals (out of 20) died, exhibiting irritation of their upper respiratory tract.
These animals did not gain as much weight as controls, exhibited decreased organ weights and
multifocal to generalized pulmonary fibrosis and inflammatory cell infiltrate. Exposure to 0.25
mg/l also resulted in an irritation of the upper respiratory tract. All treated animals survived and
their body weight gain did not differ from controls. Neither significant effects on organ weights,
nor significant effects on hematologic or biochemical parameters in the 0.25, 0.025 or 0 mg/l
treatment groups were found. Exposure to the test material at all levels resulted in an increase in
the frequency of pulmonary fibrosis and inflammatory cell infiltrate.
No compound related gross lesions were seen in any of the rats from the test groups that were
terminally sacrificed or died during the study.
Compound-related microscopic lesions, consisting of an increase in the intensity and extent of
inflammatory cell infiltrate and an increase in the incidence, intensity and extent of interstitial
fibrosis in lungs of rats from all test groups, were observed.
Ref.: 44
15
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3.3.5.2. Sub-chronic (90 days) oral / dermal / inhalation toxicity
Benzoic acid
50 male and 50 female mice received 80 mg benzoic acid/kg bw/d by gavage for 90 days. 14
surviving mice were subjected to a restricted dietary intake (90 % restriction) for up to 5 days.
10 mice were tested on their tolerance to CCl4 (test on detoxifying capacity of liver) by giving
0.1 ml CCl4 in an single dose by oral intubation at the end of the test.
Reduced body weight gain was observed and the mortality was higher in connection with
reduced tolerance to CCl4 or food restriction than in the control group. The authors did not
mention whether this finding was statistically significant. The relevance of the study will be
compared to other long-term and carcinogenicity studies.
Data are not sufficient to justify conclusion.
Ref.: 45
Sodium benzoate
Groups of 4-5 male and 4-5 female rats received 0, 1, 2, 4, and 8 % sodium benzoate equivalent
to 640, 1320, 2620, and 6290 mg/kg bw/d in the diet for 90 days.
4/8 animals died (average 13 days to death) in the 8 % dose level group, the average weight gain
of the surviving rats was reduced and the relative liver and kidney weight was significantly
increased with histopathological changes in liver and kidney (7/16).
Data are insufficient to justify a NOAEL of 4 % in the diet (2.6 g/kg bw/d).
Ref.: 8
3.3.5.3. Chronic (> 12 months) toxicity
Benzoic acid
25 male and 25 female mice were given 40 mg/kg bw/d for 17 months. Benzoic acid was fed in a
paste prior to the main feed.
The weight of liver, kidney and testes relative to body weight in mice sacrificed at the end of the
test period were lower in the group receiving sorbic acid (40 mg/kg bw/d) than in the group
treated with benzoic acid. No further details were given.
Ref.: 45
10 male and 10 female rats received 40 mg benzoic acid/kg bw/d for 18 months. Benzoic acid
was fed in a paste prior to the main feed.
The rats developed some tolerance to a lethal dose of benzoic acid given terminally (25 %
mortality after 4000 mg/kg bw compared to 100 % mortality on the control group given one dose
of 3600 mg/kg bw). No further details were given.
Ref.: 45
3.3.6. Mutagenicity / Genotoxicity
Bacterial Reverse Mutation Test
Benzoic acid
Benzoic acid (up to 10 mg/plate) was tested in the Salmonella/microsome test using S.
typhimurium TA 92, TA 94, TA 98, TA 100, TA 1535 and TA 1537. No significant increases in
the numbers of revertant colonies were detected in any S. typhimurium strains at the maximum
dose.
16
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Ref.: 29
Benzoic acid (up to 10 mg/plate) was tested in the Salmonella/microsome test using S.
typhimurium TA 97, TA 98, TA 100, TA 1535, and TA 1537 with and without metabolic
activation. Benzoic acid was non-mutagenic in this test.
Ref.: 30
Benzoic acid was tested in the Salmonella/microsome test using S. typhimurium TA 98, TA 100,
TA 1535, and TA 1537 with and without metabolic activation. Benzoic acid was nonmutagenic
in this test (< 0.0099 revertants/nmol).
Ref.: 31
Sodium benzoate
Sodium benzoate (up to 3.0 mg/plate) was tested in the Salmonella/microsome test using S.
typhimurium TA 92, TA 94, TA 98, TA 100, TA 1535 and TA 1537. No significant increases in
the numbers of revertant colonies were detected in any S. typhimurium strains at the maximum
dose.
Ref.: 29
Sodium benzoate was tested in the S. typhimurium strains TA98, TA100, TA1535, TA1537 and
TA1538 in the presence and absence of S9 mix. Sodium benzoate was non-mutagenic in this
test.
Ref.: 32
In vitro mammalian chromosome aberration test in human lymphocytes

Benzoic acid
Chromosome aberration test was carried out on benzoic acid (up to 1.5 mg/ml) using Chinese
hamster lung fibroblasts. No metabolic activation system was applied. 8 % of the cells treated
with benzoic acid (1.5 mg/ml) showed structural aberrations at 48 hrs after treatment.
Ref.: 33
Benzoic acid (1.5 mg/ml in DMSO) gave equivocal results using Chinese hamster lung
fibroblasts. 8% chromosome aberration and 1% polypoidy were reported.
Ref.: 29
Sodium benzoate
Human embryonic lung cells (WI-38) were used. Dose levels for sodium benzoate were 0, 2, 20,
200 mg/ml. 200 mg/l, previously established as the lowest toxic level. The positive control was
0.1 mg/ml triethylene melamin. The test compound was added to three culture bottles for each
dose level, 24 hours after plating. When sufficient mitoses were observed, usually after 24-48
hours, the cells were harvested and prepared for the analysis. 100 anaphases per dose were
analysed. For the mitotic index at least 500 cells were counted.
Sodium benzoate produced no significant chromosomal aberrations in human tissue culture cells
when tested at any dose level tested. No substance related effects were observed. The
chromosome abnormalities as well as the mitotic indices were within normal values.
Ref.: 34
Chromosome aberration test was carried out on sodium benzoate (up to 2.0 mg/ml) using a
Chinese hamster fibroblast cell line. No metabolic activation system was applied. 38 % of the
cells treated with sodium benzoate showed chromosome aberrations at 48 h.
17
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Ref.: 29, 35,
Chromosome aberration test was carried out on sodium benzoate using a pseudodiploid Chinese
hamster cell line (DON). No metabolic activation system was applied.
Concentration above 0.002 mol/l showed twofold background effects of chromosome
aberrations; but no increase in the frequency of sister chromatid exchange was observed.
Ref.: 36
Chromosome aberration test was carried out on sodium benzoate using human embryonic lung
culture cells. Sodium benzoate produced no significant increase in the aberration frequency in
the anaphase chromosomes when tested at the dosage levels 0, 2.0 µg/ml, 20 µg/ml and 200
µg/ml.
Ref.: 33
In vitro Sister Chromatid Exchange (SCE)

Benzoic acid
Benzoic acid, without metabolic activation, was reported to be negative for SCEs in vitro using 3
different cell lines. These were literature citations with no other information.
Ref.: 37, 38, 39
In vivo Host-Mediated Assay

0, 50, 500 and 5000 mg sodium benzoate/kg bw was given orally to mice (single dose or once a
day for 5 days) in an Host-Mediated Assay. Elevated mutant frequencies were seen with
Salmonella TA 1530 in the acute intermediate dose level. The subacute and the other acute dose
levels showed no increase in mutant frequencies. Tests with Salmonella G 46 were negative
while giving slightly elevated mutant frequencies. Tests with Saccharomyces D 3 produced no
increases in recombinant frequencies.
Ref.: 33
In vivo chromosome aberration test

Chromosome aberration of sodium benzoate was investigated in vivo in rats. Five rats per group
were dosed with 0, 50, 500 and 5000 mg/kg bw by gavage in an single dose or once a day for 5
days. The animals were killed at 6, 24 and 48 h after dosing in the acute study and 6 h after
dosing in the subacute study. Bone marrow metaphase chromosomes were checked for
aberrations.
A low incidence of treatment related breaks was observed in the acute study but within the
control range. In the subacute study, only the mid dose showed breaks (1%) but were not
considered to be significant. The mitotic indices were not different from controls. Sodium
benzoate did not induce chromosomal aberrations in this test system.
Ref.: 34
In vivo dominant lethal assay

A dominant lethal assay was conducted in rats. Following dosing with sodium benzoate by
gavage (0, 50, 500, 5000 mg/kg bw, single dose or once a day for 5 days) treated male rats, 5 per
group, were mated with two females per week for 8 weeks (acute study) or 7 weeks (subacute
study). Fertility Index, number of implantations, corpora lutea, pre-implantation losses,
resorptions/pregnant female and proportions of females with one or more dead, and two and
more and overall dead implants were monitored.
18
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In the acute study all three doses showed at week 8 significant, dose-related decreases in corpora
lutea, at week 7 significant dose-related increases of average pre-implantation losses. Average
resorptions were significant, dose-related increased at the low and high doses of week 2 and the
low and intermediate doses of week 7 were significantly increased over the control group.
Overall dead implants were significant increased at week 7 for the low and intermediate doses
and week 2 for the low dose.
In the subacute study significant increases of average pre-implantation losses were observed at a
number of weeks but no significant increases of average resorptions were observed. Overall dead
implants were not increased.
The authors considered sodium benzoate to be non-mutagenic in rats in this test system although
positive results were obtained.
Ref.: Submission 1, 33
Remark
IPCS CICAD 26 (2000) mentioned this dominant lethal assay as a positive result, however
evaluation of the raw data in the original report (by experts of the industry consortium and a
recent independent review by Prof. R. Kroes) gives no support for this. In addition the authors of
the study clearly conclude negative. FDA also evaluated this study as negative. In addition
sodium benzoate doesn’t contain a structural alert for genotoxicity.
Ref.: 55
Other recent Evaluations of Genotoxicity
OECD SIDS Initial Assessment Report on Benzoates: Benzoic acid, Sodium benzoate,
Potassium benzoate, Benzyl alcohol
They concluded that Benzoic acid and Sodium benzoate, (also Potassium benzoate and benzyl
alcohol) showed no mutagenic activity in in vitro Ames tests. Various results were obtained with
other in vitro genotoxicity assays. Sodium benzoate (and benzyl alcohol) showed no
genotoxicity in vivo. While some mixed and/or equivocal in vitro chromosomal/chromatid
responses have been observed, no genotoxicity was observed in the in vivo cytogenetic,
micronucleus, or other assays. The weight of the evidence of the in vitro and in vivo genotoxicity
data indicates that these chemicals are not mutagenic or clastogenic. They also are not
carcinogenic in long-term carcinogenicity studies.
They also remarked since the sodium salt of benzoic acid instantaneously dissociates to the
benzoic acid, the studies with sodium benzoate are also representative for benzoic acid and
potassium benzoate.
Ref.: 56
JECFA report, 1998

In addition data from in vivo genotoxicity studies on benzyl acetate and benzaldehyde are
supportive evidence for the non-genotoxicity of benzoic acid and sodium benzoate.
Ref.: 52
Scientific Committee on Food Opinion: Re-evaluation of genotoxicity

In view of the additional genotoxicity data now available, the Committee has compiled its own
summary of the studies (Annex 2). The Committee agrees that since benzylacetate,
19
SCCP/0891/05
benzylalcohol and benzylaldehyde are all metabolised to benzoic acid, the results of in vivo tests
performed with these compounds as well as with sodium benzoate may be applied also to
benzoic acid itself. Considering the database as a whole, weak genotoxic effects have been
reported mainly at chromosome level in some in vitro systems. However, all the in vivo
genotoxicity tests were negative at somatic or germ cell level. The essentially negative results
obtained in three carcinogenicity studies (one in mice, two in rats) on sodium benzoate,
nothwithstanding some limitations, give further reassurance. On this basis, it is very unlikely that
benzoic acid would interfere with chromosomes in vivo.
Ref.: 3
3.3.7. Carcinogenicity
Benzoic Acid:
20 male and 20 female rats were fed 1 % Benzoic Acid (approx. 700 mg/kg bw/d) in diet for a
maximum of 1000 days. No evidence of carcinogenicity was seen upon microscopic examination
of all analysed tissues.
Ref.: 48
Sodium benzoate
Sodium benzoate was given in 2 % in drinking water to 50 female and 50 male Swiss albino
mice from weeks 5 on for lifespan. The average daily intake of sodium benzoate was 119.2 mg
for a female and 124.0 mg for a male (approx. 5.95 - 6.2 g/kg bw/d).
There was no effect of the survival of the treated mice when compared with the untreated
control. There were no significant differences between the tumour distribution in sodium
benzoate-treated and untreated control mice.
Ref.: 42
Groups of 50 males and 52 female Fischer 344 rats were fed sodium benzoate at 1 or 2 % in the
diet (approx. 0.5 or 1.0 g/kg bw/d) for 18 to 24 months. The control group consisted of 25 males
and 43 females.
No clinical signs directly attributed to sodium benzoate were observed in treated animals.
Differences in the average body weight between the treated and control groups were negligible.
40 rats died during the first 16 months, except for myeloproliferative disorder developed in one
female control rat, all other dead animals showed pneumonia with abscess. Around 100 rats
including those of the control group died after 16 months of hemorrhagic pneumonia with
oedema.
The poor survival in all groups does limit the value of this study, although the type of tumours
was similar between test and control rats of each sex.
Ref.: 42
20
SCCP/0891/05
3.3.8. Reproductive toxicity
3.3.8.1 4-Generation reproductive toxicity
Benzoic acid
A four generation study with benzoic acid was conducted in rats. Males and females of the first
and second generation were fed 0.5 or 1.0 % benzoic acid in the diet (approximately 0.25 or 0.5
g/kg bw d). The third generation was treated for 16 weeks and generation 4 was treated until
breeding.
There were no unfavourable side-effects on growth, food utilisation, duration of life, procreation,
feeding of the offspring, weight of organs and histological pattern of organs in the 1 % dose
group. In the 0.5 % group there was a significant prolongation of lifetime.
NOAEL: 500 mg/kg bw.
Ref.: 41
3.3.8.2. Teratogenicity
Benzoic acid
In a study to determine the teratologic effects, benzoic acid was administered in a single dose of
510 mg/kg bw to 7 on pregnant Wistar albino rats at day 9 of gestation. The malformations and
resorption rates were comparable to those in control animals. The data was considered
inadequate, since only a single dose was used.
Ref.: 46
Sodium benzoate
Sodium benzoate at doses of 0, 1.75, 8, 38 and 175 mg/kg bw was administered by gavage to
groups of at least 20 pregnant albino CD outbred mice and White albino rats on gestation day 6
to 15. Groups of 21 to 22 pregnant hamsters were dosed with 0, 3, 14, 65 or 300 mg/kg bw on
gestation days 6 to 10. Groups of 10 Dutch-belted rabbits were artificially inseminated and then
dosed by oral intubation with 0, 2.5, 12, 54 or 250 mg/kg bw on gestation days 6 to 18.
Caesareans were performed on mice, rats, hamsters and rabbits on days 17, 20, 14 and 29,
respectively.
There was no clearly discernible effect on nidation or on maternal or foetal survival. The number
of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the
number occurring spontaneously in the sham-treated controls.
The NOEL in these studies was in all species identical with the highest dose tested:
mice and rats NOEL : 175 mg/kg bw

hamsters NOEL : 300 mg/kg bw
rabbits NOEL : 250 mg/kg bw
Ref.: 47
21
SCCP/0891/05
3.3.9. Toxicokinetics
Benzoic acid and its sodium and potassium salt are considered with benzylacetate,
benzylalcohol, benzaldehyde as a single category from the human health view by JECFA, as
they are all rapidly metabolized and excreted via a common pathway within 24hrs.
Sodium benzoate is expected to immediately dissociate and form benzoic acid in an aqueous
environment.
Ref.: 52
3.3.10. Photo-induced toxicity
No data was submitted.

In the JECFA report, there was a summary of an in vitro test by Eberlein-Konig et al, 1993.
Suspensions of human erythrocytes were incubated with Benzoic Acid and Sodium Benzoate.
Each material was tested at 10-5, 10-4 and 10-3 mol/l. Erythrocyte-free samples were also
incubated with the test material and used as controls. Following incubation, suspensions and
samples were exposed to varying amounts of UVA Iight from one of three sources. Hemolysis
was measured as a function of absorbance of 550 nm light.
The substances did not produce significant photohaemolysis.
Ref.: 52
3.3.11. Human data
Included in the appropriate sections
22
SCCP/0891/05
3.3.12. Special investigations
3.3.13. Safety evaluation (including calculation of the MoS)
100% skin absorption was assumed since the dermal absorption studies were old and not to
modern standards.
CALCULATION OF THE MARGIN OF SAFETY
Rinse off-products (ro):

Maximum content of active ingredient (a. I.) [%] 2.5
Exposure to rinse-off-products E [g/day] 0.72
Maximum amount of a. I. applied Iro [mg/day] 18.00
Oral hygiene-products (oh):
Exposure to oral hygiene-products E [g/day] 3.52
Maximum amount of a. I. applied Ioh [mg/day] 59.84
Eye-products (ep):
Exposure to eye-products E [g/day] 0.05
Maximum amount of a. I. applied Iep [mg/day] 0.25
Non rinse off-products (nro):
Exposure to non rinse-off-products E [g/day] 13.50
Maximum amount of a. I. applied Inro [mg/day] 67.50
Typical body weight (human) TBW [kg] 60 kg
Exposure from all product groups:

Systemic exposure dose SED [mg/kg bw/d] = [Ioh + Iro + Iep + Inro x A]/TBW = 2.43
No observed adverse effect level (NOAEL) from a 4-generation study 500 mg/kg bw/d
(selected by SCF)
MARGIN OF SAFETY (MOS):
MOS (NOAEL/SED) 500 mg/kg bw/d / 2.43 mg/kg bw/d = 206
23
SCCP/0891/05
3.3.14. Discussion
Since the adoption of the previous opinion of the SCCNFP on 4 June 2002 on Benzoic acid and
Sodium benzoate (SCCNFP/0532/01, final), there have been 2 major re-evaluations of the data
on Benzoic acid and sodium benzoate.
The SIDS report agrees that many toxicological studies on benzoic acid and its salts are old and
“do not always fulfil for 100% present-day guidelines”. They did appear to be acceptable studies
for evaluation, since ‘well-known’ research groups and/or test laboratories ran the studies
according to scientific standards and or accepted protocols at that time. Also, all were peer-
reviewed and published in high quality scientific literature. Most of them have been reviewed
and accepted by other fora like FDA, JECFA, and IPCS as acceptable studies.
In addition, there is good consistency in the individual data for a substance in the group as well
as between members of the group (benzyl acetate and benzaldehyde data inclusive). Therefore,
taken as a whole, the available studies give a robust database for hazard assessment and hazard
evaluation of these compounds and further studies are not indicated. The JECFA Committee
(1997) concluded that the data reviewed for compounds in this group were sufficient to
demonstrate lack of teratogenic, reproductive or carcinogenic potential. Consequently, the
Committee concluded that further studies were not required’
Ref.: 56
The conclusions of the SCF in 2002 were:
”The database is much more extensive than that considered by the Committee in 1994, both for
developmental toxicity and for genotoxicity.
There appear to be sufficient studies to conclude absence of teratogenic potential, with an overall
NOAEL for developmental toxicity of 500 mg/kg bw/day, based on effects on foetal weight. The
fact that this overall NOAEL takes into account gavage as well as dietary studies gives further
reassurance. It is therefore concluded that a further teratogenicity study on benzoic acid should
no longer be required.
Similarly for genotoxicity, while some of the in vitro tests have been positive or equivocal, all
the results from in vivo studies have been negative. It is therefore concluded that an in vivo study
for clastogenic activity on benzoic acid should no longer be required.
On the basis of these data and the other types of study previously evaluated by the Committee,
the Committee can establish a full Group ADI of 0 - 5 mg/kg bw for benzoic acid and its salts
including benzyl alcohol and related benzyl derivatives used as flavourings.”
Ref.: 3
This consensus of opinion for teratogenic, reproductive or carcinogenic potential of benzoic acid
and sodium benzoate is welcome.
Benzoic acid and sodium benzoate rapidly metabolize and excrete via a common pathway within
24hrs. Systemic toxic effects on liver and kidney were observed.
Benzoic acid and sodium benzoate have low acute oral and dermal toxicity with LD50 values
>2000 mg/kg bw. The 4 hrs inhalation exposure of benzoic acid at 0.026mg/l/h also show low
acute inhalation toxicity.
Benzoic acid is a mild skin irritant, but sodium benzoate was not a skin irritant. Neither benzoic
acid nor benzoate gave indication of a sensitizing effect in animals, but occasionally very low
positive reactions were recorded with humans in patch tests with benzoic acid. It has been
suggested that these positive reactions are a non-immunologic contact urticaria.
24
SCCP/0891/05
In the CICAD report, the conclusion on benzoic acid and sodium benzoate was: “However, both
substances are known to cause non-immunologic immediate contact reactions. This effect is
scarce in healthy subjects, while in patients with frequent urticaria or asthma, symptoms or
exacerbation of the symptoms were observed”. This was considered to be a possible cholinergic
mechanism. This should be borne in mind for products used on children.
Benzoic acid should be labelled as an eye irritant under the EU criteria for classification.
The percutaneous absorption study of 14C-labelled benzoic acid in vitro with excised human skin
was old, not conforming to modern guidelines.. Total absorption was found to be approximately
45 %. These data were comparable to in vivo data in human, in which the total absorption of
labelled benzoic acid was approximately 43 %.
Data is only given for sodium benzoate, but not the other salts or esters. It is thought that the loss
of acidity due to the salt (sodium, potassium) should decrease toxicity. Since the loss of acidity
is not uniform, this could be a problem. No data was provided for the esters.
4. CONCLUSION
Adequate data was provided only for benzoic acid and sodium benzoate, but not the other salts
or esters.
On the basis of provided data, the SCCP is of the opinion that benzoic acid and sodium benzoate
are safe for use for preservative and non-preservative purposes in cosmetic rinse-off products at
a maximum concentration of 2.5 % and in cosmetic oral-care products at a maximum
concentration of 1.7 %, and in leave-on products up to 0.5%. The possible non-preservative
functions have not been stated.
However, in the interest of consumer safety, generation of data on irritation and sensitisation of
the other salts and esters is required if they are to be used in cosmetic products.
Comment: Inclusion of all relevant data should have been made available, particularly if from
the grey literature as required by the SCCNFP Notes of Guidance (SCCNFP/0690/03). In the
IUCLID database, more modern percutaneous absorption studies were summarised. The study
for eye irritation was not provided with the Submission I. These should all have been provided
with Submission I.
5. MINORITY OPINION
Not applicable
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SCCP/0891/05
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* Referred to but no documentation supplied.
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Substances, VUOS 539-18, Cincinnati , Ohio, USA, 29 March. Cited in Henschler D (ed) Toxikologisch-
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2. Abe S et al. (1984) Studies on the Toxicity of Oxaprozin (1) Acute Toxicity of Oxaprozin, its Metabolites
and Contaminants. IYAKUHIN KENKYU 15 359 - 70 (Japanese, only abstract and tables in English)
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Hirzel, Wissenschaftliche Verlagsgesellschaft, (1995)
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Swelling Test (MEST) Toxicol Appl Pharm 84 93 - 114
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Bayer AG data. Cited in BUA-Stoffbericht 145, December 1993. S. Hirzel, Wissenschaftliche
Verlagsgesellschaft, (1995)
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BUA-Stoffbericht 145, December 1993. S. Hirzel, Wissenschaftliche Verlagsgesellschaft, (1995)
*22. RCC NOTOX Acute eye irritation/corrosion study with natrium benzoate in rabbits (study no. 014669). RCC
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Wissenschaftliche Verlagsgesellschaft, (1995)
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(1989) TNO BIBRA Toxicology International Ltd.
24. Leyden JJ & Kligman AM (1977) Contact sensitization to benzoyl peroxide. Contact Derm 3 273 - 275
25. Broeckx W et al. (1987) Cosmetic intolerance. Contact Derm 16 189 – 194
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Chem Toxic 22 623 - 636
27. Zeiger E et al (1988) Salmonella Mutagenicity Test: IV. Results From the Testing of 300 Chemicals. Environ
Mol Mutagen 11 Suppl. 12 1 - 18
28. McCann J et al (1975) Detection of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300
chemicals. Proc Nat Acad Sci 72 5135 - 5139
29. Prival MJ et al (1991) Bacterial mutagenicity testing of 49 food ingredients gives very few positive results.
Mutat Res 260 321 - 329
28
SCCP/0891/05
30. Ishidate M JR et al (1988) A comparative analysis of data on the clastogenicity of 951 chemical substances
tested in mammalian cell cultures. Mutat Res 195 151 - 213
31. Ishidate M JR & Odashima S (1977) Chromosome tests with 134 compounds on Chinese hamster cells in
vitro - a screening for chemical carcinogens. Mutat Res 48 337 - 354
32. Abe S & Sasaki M (1977) Chromosome Aberration and Sister Chromatid Exchange in Chinese Hamster
Cells Exposed to Various Chemicals. J Natl Cancer Inst 58 1635 - 1641
33. Fabrizio DBA (1974) Mutagenic Evaluation of Compound FDA 71-27 Sodium Benzoate, Litton Bionetics,
Inc. (very bad copy!)
34. Frantz TJ (1975) Percutaneous absorption. On the relevance of in vitro data. J Invest Derm 64 190 - 195
35. Hunziker et al (1978) Animal Models of Percutaneous Penetration: Comparison between Mexican Hairless
Dogs and Man. Dermatologica 156 79 - 88
36. Toth B (1984) Lack of Tumorigenicity of Sodium benzoate in Mice. Fundam Appl Toxicol 4 494 - 496
37. Fanelli GM & Halliday SL (1963) Relative toxicity of Chlortetracycline and Sodium benzoate after oral
administration to rats. Arch Int Pharmacodyn 144 120 - 125
38. Shtenberg AJ & Ignat'ev AD (1970) Toxicological Evaluation of some Combinations of Food Preservatives.
Fd Cosmet Toxicol 8 369 - 380
39. Food and Drug Research Labs., Inc. (1972) Teratologic evaluation of FDA 71-37 (sodium benzoate) in mice,
rats, hamsters and rabbits. NTIS Report PB-221 777
40. Kimmel CA et al (1971) Studies on Metabolism and Identification of the Causative Agent in Aspirin
Teratogenesis in Rats. Teratology 4 15 - 24
41. Kieckebusch W & Lang K (1960) Die Verträglichkeit der Benzoesäure im chronischen Fütterungsversuch.
Arznei-Forsch 10 1001- 1004
42. Sodemoto Y & Enomoto M (1980) Report of carcinogenesis bioassay of Sodium benzoate in rats: Absence
of carcinogenicity of sodium benzoate in rats. J Environ Pathol Toxicol 4 87 - 95
*43. Not given
*44. Not given
45. Beratergremium für umweltrelevante Altstoffe (BUA) der Gesellschaft Deutscher Chemiker (ed) BUA-
Stoffbericht 145, December 1993. S. Hirzel, Wissenschaftliche Verlagsgesellschaft, (1995)
46. BIBRA Report Toxicity Profile - Benzoic Acid (1989) TNO BIBRA Toxicology International Ltd.
Additional references (provided by SCCNFP):
AR 1. European Parliament and Council Directive No 95/2/EC of 20 February 1995 on foodadditives other than
colours and sweeteners. Official Journal L 061 , 18/03/1995 p. 0001-0040
AR 2. Commission Directive No 96/77/EC of 2 December 1996 laying down specific purity criteria on food
additives other than colours and sweeteners. OJ No L339, 1-6.
AR 3. FDA, Code of Federal Regulations, Title 21- Food and Drugs, Volume 3 [Revised as of April 1, 2001], 184.
464-559.
AR 4. Handbook of pharmaceutical excipients 3rd ed 2000 editors: Kibbe, I, Arthur H, Pharmaceutical Society of
Great Britain; American Pharmaceutical Association
AR 5. Martindale : The Complete Drug Reference 32nd ed. 1999 editors: Parfitt K, Sweetman, S C, Blake P S,
Parsons, A V.
AR 6. Lahti A (1980) Non-immunologic contact urticaria. University of Oulu.
AR 7. Final report on the safety assessment of benzyl alcohol, benzoic acid, and sodium benzoate. Cosmetic
Ingredient Review, Washington, DC, 20036, USA. Intern. Journal of Toxicology (2001), 20 (Suppl. 3), 23-
50
AR 8. International Programme On Chemical Safety, (2000) Concise International Chemical Assessment Document
No. 26 Benzoic Acid And Sodium Benzoate. http://www.inchem.org/documents/cicads/cicads/cicad26.htm
AR9. Feldmann RJ & Maibach HI (1970) Absorbtion of some organic compounds through the skin in man. J
Invest Derm 54 399-404).
AR10. Edwards RC, Voegeli CJ (1984) Inadvisability of using caffeine and sodium benzoate in neonates. Am J
Hosp Pharm 41, 658.
AR11. Schiff D et al (1971) Fixed drug combinations and the displacement of bilirubin from albumin. Pediatrics 48,
139 -41.
7. ACKNOWLEDGEMENTS
29
SCCP/0891/05
Members of the working group are acknowledged for their valuable contribution to this opinion.
The members of the working group are:
Dr. C.M. Chambers (rapporteur)

Prof. R. Dubakiene
Dr. R. Grimalt
Dr. B. Jazwiec-Kanyion
Prof. V. Kapoulas
Prof. C. Lidén
Prof. J.-P. Marty
Dr. S.C. Rastogi
Prof. J. Revuz
Prof. T. Sanner
Dr. I.R. White (chairman)
30
JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY
Volume 4, Number 5, 1985
vary Ann Liebert, Inc., Publishers
Final Report on the Safety

Assessment of Butylene Glycol,
Hexylene Glycol, Ethoxydiglycol,
and Dipropylene Glycol
Butylene Glycol, Hexylene Glycol, Ethoxydiglycol, and Dipropylene Glycol are

viscous liquids used in the cosmetic industry as humectants, emulsifiers, plasti-
cizers, and solvents.
The results of acute, subchronic, and chronic oral toxicity studies using a
variety of animal species indicate a low order of toxicity for the Glycols. Results
of parenteral injection, inhalation, and acute and subchronic cutaneous toxic-
ity studies likewise support a low order of toxicity. Butylene Glycol, Ethoxydi-
glycol, and Dipropylene Glycol caused minimal to mild irritation of rabbit skin,
whereas Hexylene Clycol was moderately irritating. The Glycols produced mild
to severe ocular irritation when tested in rabbits, with Hexylene Clycol produc-
ing the most severe irritation. Although undiluted Hexylene Glycol produced
severe ocular irritation, a 25 percent aqueous solution produced no signs of ir-
ritation. Undiluted Butyiene Glycol was not an eye irritant to rabbits but was to
humans.
Human skin patch tests on undiluted Butylene Glycol and undiluted Hex-
ylene Glycol produced a very low order of primary skin irritation. A repeated
insult patch test on Butylene Glycol produced no evidence of skin sensitiza-
tion.
Based on the available data it is concluded the Butylene Glycol, Hexylene
Glycol, Ethoxydiglycol, and Dipropylene Glycol are safe as presently used in
cosmetics.
INTRODUCTION
B
utylene Glycol, Hexylene Glycol, Ethoxydiglycol, and Dipropylene Glycol
are viscous liquids used in the cosmetic industry as humectants, emulsifiers,
plasticizers, and solvents.
223
224 COSMETIC INGREDIENT REVIEW
CHEMICAL AND PHYSICAL PROPERTIES
Structure/Composition
1. Butylene Glycol is an aliphatic diol. It conforms to the formula:
HOCHzCH,CHCH,
CAS Number: 107-88-o
Other names include 1,3-Butanediol and 1,3-Butylene Glycol.(‘)
2. Hexylene Glycol is the aliphatic alcohol that conforms to the formula:
CH,
I
CHa-C-CHz-CHCHS
I I
OH OH
CAS Number: 107-41-5
Other names are 2-Methyl-2,4-Pentanediol and 2,4-Pentanediol,2-Methyl-.(‘)
3. Ethoxydiglycol is the ether alcohol that conforms to the formula:
CAS Number: 11 l-90-0
Other names include Diethylene Glycol Monoethyl Ether; Ethanol, 2-(2-Eth-

oxyethoxy)-; 2-(2-Ethoxyethoxy)EthanoI.(‘)
4. Dipropylene Glycol is a mixture of diols that conforms generally to the

formula:
CHLHCHz-0-CHLHCt+
I I
OH OH
The material of commerce is primarily a mixture of 3 isomers, with the majority

being disecondary (85 to 90 percent). Primary-secondary and diprimary isomers,
along with up to 5 percent unidentified material, make up the remainder.
CAS Number: 110-98-5
Other names are: Di-1,2-Propylene Glycol; 1,l ‘-Oxybis-2-Propanol; 2-Propa-

nol, 1,l ‘-Oxybis-. (L.2)
FINAL REPORT: SAFETY ASSESSMENT OF THE GLYCOLS 225
Properties
Butylene Glycol is a clear, practically colorless, viscous, hygroscopic liquid.

It is odorless and has a slightly sweet, characteristic taste, Butylene Glycol is mis-
cible in all proportions with water, acetone, and alcohol. It is immiscible with
fixed oils and insoluble in aliphatic hydrocarbons, benzene, toluene, and carbon
tetrachloride. This glycol does dissolve most essential oils and synthetic flavoring
substances.(3-7)
Hexylene Glycol is a clear hygroscopic liquid with a mild, sweet odor. It ex-
hibits exceptional solvency for a variety of materials and is miscible with aliphatic
and aromatic hydrocarbons as well as with such polar substances as water, fatty
acids, and alcohols. It is combustible.(5*6*s)
Ethoxydiglycol is a colorless hygroscopic liquid, soluble in water and most
organic solvents.(4vg)
Dipropylene Glycol is a colorless, slightly viscous liquid that is soluble in
water, ethanol, and acetone.(5)
Other physical and chemical properties of the glycols are shown in Table 1.
Production
Butylene Glycol is produced by the catalytic hydrogenation of acetaldehyde

using as catalysts Raney nickel, copper, or platinum oxide. It is purified by distil-
lation. (6.7.lO)
TABLE 1. Properties of Glycols
Butylene Hexylene Ethoxydi- Dipropylene

Property Clycol Glycol glycol Clycol Reference
Molecular weight 90.12 188.18 134.18 134.18 4
Boiling point (“C) 208 (0.8 atm) 1g7vmmm) 195V60mm) 229-32 4
207.5 197.1(7f3hm) - 222-38(760mm) 2, 7, 8
Freezing point (T) -50 - -105 Supercools 2, 6, 7, 9

Specific gravity (g/ml) 1.0053': 0.9254': 0.9881': 1.0224:: 4
1.0053:: 0.9233:: 0.988:: 1.020-1.030D 2, 7-9
Refractive index r$ 1.4418 1.4250 1.4300 - 4
1.4412 1.4276 - 1.439 2, 7, 8
Vapor pressure 0.06 mm - 0.26 mm 0.01 mm 2, 4, 5, 9

Viscosity (cps) 103.9 (25°C) 41.7 (209C) - - 7,8
Solubility
Water Soluble Soluble Soluble Soluble 3, 4
Alcohol Soluble Soluble Soluble Soluble 3, 4
Ether Insoluble Soluble Soluble - 4
Acetone Soluble Soluble Soluble 2, 4, 7
-
Benzene Insoluble Soluble - 4, 7
-
Carbon tetrachloride Insoluble - - - 7
Aliphatic hydrocarbons Insoluble Soluble - 7,8
-
Aromatic hydrocarbons - Soluble - - 8
Fatty acids - Soluble - 8
-
Hexylene Glycol is manufactured by the condensation of 2 molecules of ace-

tone to produce diacetone alcohol, which is further hydrogenated to produce
Hexylene Glycol. This is then purified by distillation.‘*’
Ethoxydiglycol is prepared from the reaction of ethylene oxide with ethanol
followed by purification by distillation.‘g’
Dipropylene Glyol is the reaction product of propylene glycol and propylene
oxide.(‘sl’)
Analytical Methods
Chemical and chromatographic methods may be used for the identification

and separation of the glycols. Chemical methods involve oxidation of the glycols
with subsequent reaction with a reagent. (I*) Gas chromatography may be used
for the identification of gIycoIs.“3,14’
Impurities
Butylene Glycol has a minimum gas chromatographic assay of 99.5 percent

by weight. It contains a maximum of 0.5 percent water, up to 0.005 percent
acetic acid, and trace amounts of branched 1,3-Butylene Glycol.“)
Hexylene Glycol contains a maximum of 0.1 percent water and 0.005 per-
cent acetic acid. (8)
Ethoxydiglycol has a minimum gas chromatographic assay of 99.0 percent
and contains water and acetic acid to maximums of 0.1 and 0.01 percent, respec-
tively. Known impurities include diethylene glycol and triethylene glycol.(g)
The isomer distribution of Dipropylene Glycol is as specified by the commer-
cial buyer. It contains up to 0.1 percent water, up to 0.01 percent acetic acid, and
up to 0.001 percent inorganic chlorides. The maximum combustion residue of
Dipropylene Glycol should not exceed 0.005 percent.(*)
USE
Noncosmetic Use
Butylene Glycol has been tested as a parenteral drug solvent,“” in the manu-
facture of polyester plasticizers, and as a humectant for cellophane and tobac-
CO.(~) It serves as a surfactant, coupling agent, and solvent.“) Butylene Glycol has
both indirect food additive (IFA) and direct food additive (DFA) status with the
Food and Drug Administration when used as a solvent for flavors,“” as an anti-
oxidant and stabilizer, and as a component of packaging (21 Code of Federal
Regulations [CFR] 173.220; 177.1200; 175.105; 177.1680; 177.1210;
178.2010).'16'
Hexylene Glycol has IFA status as a defoaming agent (21 CFR 176.210).(‘6) It
is also used in hydraulic brake fluids, printing inks, and textile dye vehicles. Hex-
ylene Glycol serves as a coupling agent, as a fuel and lubricant additive and as an
emulsifying agent.“.“)
FINAL REPORT: SAFETY ASSESSMENT OF THE CLYCOLS 227
Exthoxydiglycol has IFA status for use as a component of paperboard and ad-
hesives (21 CFR 175.105; 176.180).““’
Dipropylene Glycol is an IFA ingredient for use in adhesives, lubricants, and
as a defoaming agent (21 CFR 175.105; 178.3910; 1 76.200).“6’ It is also used as a
solvent for nitrocellulose and shellac and as a partial solvent for cellulose acetate.
Other applications include solvent mixtures, lacquers, coatings, and printing
inks.“’
Cosmetic Use
The glycols generally are used as cosmetic emulsifiers, solidifiers, plasti-

cizers, cosolvents, and film producers. (“.l*) Butylene Glycol is used as a humec-
tant, especially in hair sprays and setting lotions. (l*-*‘) It helps retard the loss of
aromas from essential oils, preserves against spoilage by microorganisms, and is
used as a solvent for benzoates. Special grades of Dipropylene Glycol (based on
odor quality) are used as carriers for perfume oils.(*)
Cosmetic product formulation data on the use and occurrence of the glycols
in finished products are made available by the Food and Drug Administration
(FDA) and compiled through voluntary filing of such data in accordance with
Title 21 part 720.4 of the Code of Federal Regulations. Ingredients are listed in
prescribed concentration ranges under specific product type categories. Since
certain cosmetic ingredients are supplied by the manufacturer at less than 100
percent concentration, the value reported by the cosmetic formulator may not
necessarily reflect the true concentration found in the finished product; the
actual concentration in such a case would be a fraction of that reported to the
FDA. Since data are only submitted within the framework of preset concentration
ranges, the opportunity exists for a 2- to lo-fold overestimation of the actual con-
centration of an ingredient in a particular product (Table 2).(*l)
Butylene Glycol is used in 165 products at concentrations from less than 0.1
percent to greater than 50 percent. It is used in hair and bath products, eye and
facial makeup, fragrances, personal cleanliness products, and shaving and skin
care preparations.‘*”
Hexylene Glycol is used in 85 cosmetic formulations at concentrations of use
ranging from less than 0.1 percent to 25 percent. The types of products in which
Hexylene Glycol is used include bath and hair preparations, eye makeup, soaps,
and skin care preparations.‘*”
Ethoxydiglycol has been reported as an ingredient in 80 product formula-
tions at concentrations from less than 0.1 percent to 50 percent. The types of
products include eye makeup, fragrances, hair and nail preparations, and shav-
ing and skin care preparations.‘*”
Dipropylene Glycol is reported as an ingredient in 50 cosmetic formulations
in concentrations ranging from less than 0.1 percent to 50 percent. It is used in
hair care and bath products, perfumes, facial makeup, doedorants, and shaving
and skin care preparations.‘*l’
The Glycols may contact all parts of the integument to which the products
are applied and may remain in contact for several hours daily.‘*‘)
TABLE 2. Product Formulation Data’*‘)
No. Product Formulation5

Total No. Within Each Concentration Range I%)
Containing
Product Category* Ingredient >50 > JO-25 >5-70 >J-5 >O.l-l 50. I
Butylene Clycol
Bath oils, tablets, and salts 1 - - - -

Other bath preparations 4 1 - - -
Eyeliner 3 - - 3 -
Eye shadow 13 - 12 - -
Eye makeup remover 4 - - 4 -
Mascara 34 - - 29 -
Other eye makeup prepara- 1 - - 1 -
tions
Colognes and toilet waters - 1 - - 2
Perfumes - - - 2 -
Other fragrance preparations - - 1 - -
Hair conditioners - - 2 1 1
Hair shampoos (noncolor- - - - - -
in@
Tonics, dressings, and other 1 - - - 1 -
hair grooming aids
Blushers (all types) 7 - 1 3 3 -
Face powders 1 - - - 1 -
Makeup foundations 19 - 3 16 - -
Makeup bases 1 - - 1 - -
Rouges 2 1 - 1 - -
Other makeup preparations 2 - 1 1 - -
(not eye)
Cuticle softeners - - 1 - -
Bath soaps and detergents - - 1 - -
Deodorants (underarm) - 1 - - -
Other personal cleanliness - - - 1 -
products
Aftershave lotions 4 - - - 2 2
Skin cleansing preparations 13 - - 2 10 -
(cold creams, lotions,
liquids, and pads)
Face, body, and hand skin 8 - 1 2 2 1
care preparations (exclud-
ing shaving preparations)
Moisturizing skin care prep 11 - 3 2 6 -
arations
Night skin care preparations - - - - -
Paste masks (mudpacks) - - 1 1 1
Skin fresheners - - - 4 1
Wrinkle smoothers (re- - - - 1 -
movers)
Other skin care preparations 7 - - 2 3 1
Suntan gels, creams, and 1 - - - 1 -
liquids
1981 TOTALS 165 2 23 46 76 9

TABLE 2. (Continued)
No. Product Formulations Within

Total No. Each Concentration Range (%)
Containing
Product Category* Ingredient >10-25 >5-10 >l-5 >o. I-I 50.1
Hexylene Clycol
Bath oils, tablets, and salts 4 1 3 - -

Bubble baths 3 - - 2 1
Eye makeup remover - - - 1
Hair conditioners 7 - 1 2 4
Permanent waves 1 - - -
Hair rinses (noncoloring) 1 1 - - -
Hair shampoos (noncoloring) 29 - 10 13 5
Hair dyes and colors (all types requiring 20 17 - 3 -
caution statement and patch test)
Hair bleaches - - 1 - -
Bath soaps and detergents 3 - - 3 - -
Deodorants (underarm) 2 - - - 2 -
Skin cleansing preparations (cold 4 - - 3 1 -
creams, lotions, liquids, and pads)
Face, body, and hand skin care prepa- 1 - - 1 - -
rations (excluding shaving prepara-
tions)
Moisturizing skin care preparations 3 - - 2 1 -
Paste masks (mudpacks) - 1 - - -
Skin fresheners 3 - - 1 2 -
Other skin care preparations - - 1 - -
1981 TOTALS 85 20 15 32 17 1
No. Product Formulations

Total No. Within Each Concentration Range (%I
Containing
Product Category* Ingredient >25-50 >lO-25 >5-10 >l-5 >0.7-l 50. I
fthoxydiglycol
Mascara - 1
Colognes and toilet waters - 1
Hair conditioners 2 2
Hair shampoos (noncolor- - 1
ing)
Wave sets - - - 1 -
Hair dyes and colors (all 14 - - 4 10 -
types requiring caution
statement and patch test)
Hair tints 13 - - -
Hair bleaches 5 - - -
Other hair coloring prepa- - - -
rations
Nail polish and enamel - - 1
remover
Aftershave lotions 2 - - - - 2
Skin cleansing preparations 14 1 - 1 8 3
liquids, and pads)
TABLE 2. (Continued)

Total No. Within Each Concentration Range f%)
Containing
Product Category* Jngredient >25-50 > IO-25 >5-10 >l-5 >O.T-I 50.1
Face, body, and hand skin 3 - - - 1 1 1

Moisturizing skin care prep 3 - 2 1 -
arations
Night skin care preparations - - 1 -
Paste masks (mudpacks) 1 - 1 1
Skin lighteners - - 1 -
Skin fresheners - - 2 -
Other skin care preparations - - 1 3 1
1981 TOTALS 80 1

Total No. Within Each Concentration Range I%)
Containing
Product Category* Ingredient >50 > IO-25 >5-JO >l-5 >o. I-I so. 1
Dipfopylene Clycol
Bath oils, tablets, and salts 1 1 - - - -

Colognes and toilet waters 2 - - 2 - -
Perfumes 12 6 4 - 1 1
Sachets 1 - - - -
Other fragrance preparations 1 - - - -
Hair sprays (aerosol fixa- - - -
tives)
Hair shampoos (noncolor- 1 - -
ing)
Tonics, dressings, and other - 1 - - -
hair grooming aids
Wave sets 4 - -
Lipstick - - 1 1 -
Makeup bases - 1 -
Deodorants (underarm) - 4 -
Aftershave lotions - 1 1
Skin cleansing preparations - - -
liquids, and pads)
Face, body, and hand skin 3 - 3 -
Foot powders and sprays - - - - 1
Moisturizing skin care prep- 3 - - 1 2 -
arations
Skin fresheners 2 - 1 - - 1
Wrinkle smoothers (re- - - - 1 -
movers)
Other skin care preparations 1 1 .- -
1981 TOTALS 50 9 6 10 14 4
*Preset product categories and concentration ranges in accordance with federal filing regulations (21 CFR
720.4).
BIOLOGICAL PROPERTIES
Absorption, Metabolism, and Excretion
Romsos and associates(**) investigated the effects of Butylene Glycol on lipid

metabolism in rats, pigs, and chicks. The animals were fed a basal high carbohy-
drate diet that contained approximately 20 percent protein and 5 percent fat. In
the test diets, Butylene Glycol was substituted isocalorically for the carbohydrate.
The diets were offered ad lib except for 1 experiment in which pigs were fed to
satiety twice daily. Controls received the basal diet. Addition of up to 20 percent
Butylene Glycol to the diet did not affect body weight gain of the 3 species. Blood
fi-hydroxybutyrate content increased in all 3 species. Plasma triglyceride concen-
trations decreased in the rat, increased in pigs, and remained the same in chicks.
Plasma glucose decreased in the rat and remained stable in pigs and chicks. Die-
tary Butylene Glycol decreased the rate of fatty acid synthesis in the liver of rats,
but there was no such effect in pigs or chicks.
Mehlman et al.(23) studied the metabolic fate of Butylene Glycol in the rat.
Two groups of 14 rats each were fed for up to 7 weeks either a control diet of 70
percent carbohydrate and 30 percent fat or 45 percent carbohydrate, 30 percent
fat, and 25 percent Butylene Glycol. Body weight gain and epididymal fat pad
weight decreased in test animals receiving the test diet. Blood acetoacetate and
fl-hydroxybutyrate concentrations were increased significantly. Blood pyruvate
concentration was decreased significantly in animals fed this glycol for 7 weeks.
The metabolism of glucose and Butylene Glycol to ketones by hepatic tissue
taken from test animals was also studied. The conversion of glucose to lactate
and pyruvate was decreased, as was the concentration of ketones. In liver slices,
Butylene Glycol was metabolized to acetoacetate and P-hydroxybutyrate. Butyl-
ene Glycol, therefore, is metabolized in the cytosol and converted by the liver to
ketones; it is then oxidized in the tricarboxylic acid cycle.
Tate et al.(24) also studied the metabolic fate of Butylene Glycol in rats. They
found that the conversion of the glycol to /3-hydroxybutyrate in the liver was de-
pendent on NAD’ and inhibited by pyrazole. They found that hepatic alcohol
dehydrogenase was the catalyst in the catabolism of the glycol to an intermediate
aldol and then to /3-hydroxybutyrate.
Mehlman et al.(25) also reported that hepatic alcohol dehydrogenase was the
enzyme responsible for the initial oxidation of Butylene Glycol.
The metabolites of Butylene Glycol in the brain of rats were determined by
Morris et al.(26) The animals were fed diets containing 1 of the following: 47 per-
cent dietary calories as glucose, 47 percent calories as Butylene Glycol, or 47
percent calories as ethanol for 62 days. The animals were killed and metabolites
in the brain were determined. Butylene Glycol significantly decreased glutamate,
lactate, and pyruvate concentrations. Glucose concentrations and the NADPl
NADPHIDH ratios were also decreased.
Rats and mice excreted up to 40 percent of a 200 mg daily oral dose of Hexyl-
ene Glycol in the urine. (*‘) An oral 1 mmol/kg dose of Hexylene Glycol adminis-
tered to rabbits was excreted as glucuronate (67 percent of original dose).(28)
When administered orally or by subcutaneous injection to rabbits, Ethoxydi-
glycol was oxidized in the body or excreted as glucuronate. Such administration
was followed by a marked increase in the urinary content of glucuronic acid.(2g)
-
Reproductive Physiology
The effect of the ingestion of Butylene Glycol on pregnant rats and on the
metabolism of their offspring was studied. r30) Groups of pregnant rats were given
water (control) or Butylene Glycol (9 percent w/v) in drinking water. Treatment
was continued throughout the period of gestation and lactation. The length of
gestation was not affected by the glycol. The investigators found that rats that in-
gested Butylene Glycol through gestation bore offspring with a slight increase in
RNA content in neurones taken from the cerebral cortex at 18 days. In 8- and
18-day-old pups, protein synthesis in the liver was significantly reduced. In
amino acid incorporation studies, neuronal perikarya from 8-day-old pups incor-
porated 45 percent more amino acids into acid-insoluble polypeptides than did
controls. However, protein synthesis in neurons from 18-day-old pups was se-
verely inhibited by maternal ingestion of the glycol. Therefore, maternal treat-
ment with the glycol exerted opposite effects on neuronal protein synthesis at dif-
ferent stages of postnatal development of progeny. Amino acid incorporation by
free and membrane-bound ribosomes from liver of 8- and 18-day-old pups was
increased by Butylene Glycol.
Neuropharmacology and Behavior
Ayers and lsgrig (31) studied the effect of Butylene Glycol on the behavior of
rats in several experiments. To study its effect on voluntary activity (running), the
compound was administered intragastrically in a 3.5 g/kg dose, once a week for 3
weeks. Glycerol, water, sucrose, or a sham was administered on 4 other days in
the week. Butylene Glycol depressed voluntary running activity; this finding
might be due to the glycol’s interference with hunger motivation, since intuba-
tion of the compound depressed food and water intake. These investigators also
studied the dose-effect of Butylene Glycol on food and water intake and urine
output in several different experiments. In one study, groups of 8 male Charles
River Sprague-Dawley rats were fed Butylene Glycol in doses of 0, 1.75, 3.5,
5.25, or 7.0 g/kg. Corn oil was added to the last 4 dosages to produce a constant
caloric value. Distilled water and lecithin (emulsifier) were also added to pro-
duce a constant dose volume of 11.4 ml/kg. Each dose was administered to each
animal daily over a 5-day period. Ingestion of the glycol produced a dose-related
depression of activity and food and water intake. In another experiment, 10 male
Charles River Sprague-Dawley rats were trained to balance on a rotating dowel.
Butylene Glycol (7.0 g/kg) was administered once a week for 3 weeks. Other ani-
mals received glycerol, sucrose, or ethanol. The glycol produced more falls than
any of the other test compounds. The glycol-treated rats were barely able to
stand, and Butylene Glycol apparently acted as a CNS depressant or a muscle re-
laxant.
The effects of Butylene Glycol on neuropharmacology, behavior, and CNS
function were studied in male and female Sprague-Dawley rats, which were
given IP doses of 0.2 g/ml of Butylene Glycol in sterile 0.15 M saline.‘32’ Control
animals were given equal volumes of saline. Another group of rats was fed a
liquid diet containing Butylene Glycol in increasing concentrations (0.07 g/ml for
2 days; 0.08 g/ml for 2 days; 0.09 g/ml for 3 days; then 0.1 g/ml for 5 days). The IP
administration of the glycol caused a dose-related impairment of motor coordina-
tion 1 hour after treatment as measured by aerial righting reflex. Administration

of the compound depressed cGMP content in the cerebellum but did not alter
plasma-leuteinizing hormone. “Conflict behavior,” as measured by the number of
electrical shocks accepted by rats, was attenuated after treatment with glycol,
i.e., more shocks were accepted by treated rats than by control rats. Compound
treatment also caused a decrease in blood pH and blood pressure. When etha-
nol-withdrawn rats were acutely treated with Butylene Glycol, a dose-related de-
crease in tremors was observed. Feeding of rats caused no significant motor coor-
dination impairment, and all rats gained weight. After 12 days, the glycol was
removed from the diet. No tremors developed after 1 hour, but 1 of 9 rats devel-
oped seizures. After 7.5 hours, tremors increased, and 33 percent of the rats de-
veloped seizures; 1 died.
Microbiological Effects
Butylene Glycol can be used as the sole carbon source by some strains of my-
cobacteria.‘““) However, other investigators found it to be toxic to some microor-
ganisms and useful as a cosmetic preservative.(lg) Harb and Toama’34) reported
that Butylene Glycol is the most efficient polyol as an antimicrobial agent. It in-
hibits both gram-positive and gram-negative microorganisms, as well as molds
and yeast. However, it is not sporicidal. Those microbes against which Butylene
Glycol is effective include Escherichia co/i, Pseudomonas aeroginosa, Staphylo-
coccus aureus, Corynebacterium hofmanii, Aspergillus niger, Aspergillus fumi-
gatus, Pityrosporum oxalicurn, Fusarium sp., and Candida albicans.
Animal Toxicology
Oral Toxicity
Acute Studies
The acute oral LDSO of Butylene Glycol was 23 g/kg in rats(6.35) and 11 g/kg in
guinea pigs.(35’ A nail lotion containing 5.0 percent Butylene Glycol had an LDso
of > 5 g/kg in rats, (36) and a product containing 21.35 percent Butylene Glycol
produced no deaths when fed to rats at 15 g/kg.(37)
Windholz(6’ reported the acute oral LDso of Hexylene Glycol in rats was 4.70
g/kg. Other sources reported an oral LDSO of 4000 mg/kg for rats,(35) 3290 mg/kg
for rabbits,‘38’ 2800 mg/kg for guinea pigs,‘38’ and 3900 mg/kg for mice.(35) A skin
care product formulation containing 1.6 percent Hexylene Glycol was found to
be “slightly toxic” or “practically nontoxic” when administered orally to groups of
10 rats in 4 separate assays. (3g)An eye makeup remover containing 1 .O percent
Hexylene Glycol caused no deaths when administered orally to 10 mice at 15
ml/kg. c40)
The acute oral LDso values for Ethoxydiglycol are: in rats, 5.54 g/kg; in mice,
6.58 g/kg; and in guinea pigs, 3.87 g/kg. (41) A paste mask product formulation
containing 2 percent Ethoxydiglycol caused no signs of toxicity when adminis-
tered orally to 10 rats at 13 ml/kg. (42) A body lotion containing 1 .O percent Eth-
oxydiglycol caused no deaths when 15 ml/kg oral doses were administered to 10
mice. (43)
-
The acute oral LDso of Dipropylene Glycol in rats was 15 g/kg.‘35’ A shaving
preparation containing 7.2 percent Dipropylene Glycol had an oral LDso of >5
g/kg. (44)
Subchronic Studies
For subchronic effects of Butylene Glycol, see also the Metabolism section of
this report.
Larsen(*‘) reported that Hexylene Glycol fed to mice daily at 5, 10, or 20 mg
Hexylene Glycol for 57 to 81 days produced no effect on growth curves. Also,
rats receiving up to 150 mg of this glycol in the diet daily for 4 months had no ab-
normality in growth, behavior, or fertility; some changes were present in renal tis-
sue of rats of the 200 mg/day group.(*‘)
No effect was caused by the daily consumption of 0.49 g/kg Ethoxydiglycol
by each of 5 rats for 30 days. However, 0.87 g/kg caused reduced feed consump-
tion. (45)
Four groups of Charles River Wistar rats, each consisting of 12 males and 12
females, were fed diets containing 0 (control), 0.25, 1 .O, or 5.0 percent Ethoxydi-
glycol for periods up to 90 days. Observations were made of body weight, food
intake, hematological parameters at 6 and 12 weeks, kidney function, blood urea
concentration, and urine composition. Organs examined were the liver, kidneys,
brain, spleen, heart, adrenals, and gonads. The investigators found that through-
out the 90 days, the general conditions remained good. One male rat in the 5
percent group died on Day 23; there was weight loss and degeneration of renal
tubules and liver. Growth rates of male and female rats were reduced when fed
the 5 percent diet, and this was associated with a decrease in food consumption.
No hematological changes were produced by any diet. However, activity of
urinary glutamic-oxaloacetic transaminase was increased in both sexes fed the 5
percent diet, which was indicative of impaired renal function. An increase in
weight of the kidneys occurred in both sexes fed the 5 percent diet. No other im-
portant changes were found.(46)
The effect of subchronic ingestion of Ethoxydiglycol was studied in rats,
mice, and pigs. The glycol was fed for 90 days to groups of 15 male and 15 female
rats at dietary concentrations of 0 (control), 0.5, or 5 percent and to groups of 20
male and 20 female mice at dietary concentrations of 0 (control), 0.2, 0.6, 1.8,or
5.4 percent. Three groups of 4 male and 4 female pigs were fed daily oral doses of
0 (control), 167, 500, or 1500 mg/kg. There was a reduction of growth in rats and
mice at the highest concentrations. All 3 species had reduced hemoglobin con-
centration at the highest doses administered. Oxaluria occurred in rats and mice
at highest concentrations. Three pigs given 1500 mg/kg per day for up to 21 days
died with signs of uremia. For the surviving pigs, the dose was then reduced to
1000 mg/kg per day. Six of the 20 male mice fed the 5.4 percent diet died of renal
damage. The relative weights of the kidneys was increased in all 3 species fed the
highest concentration of glycol and in mice fed the 1.8 percent diet. Microscopic
changes were hydropic degeneration of the proximal renal tubules in all 3 spe-
cies fed the highest dosage and in pigs receiving 500 mg/kg per day. Hydropic de-
generation of the hepatocytes was observed in those pigs above a dose of 500
mg/kg. Hepatic cell enlargement was found in those mice fed the 1.8 and 5.4 per-
cent diets. The “no effect” level for Ethoxydiglycol in rats was 0.5 percent of the
diet. In mice it was 0.6 percent of the diet, and in pigs it was 167 mg/kg per
day.(47)
Chronic Studies
Butylene Glycol was fed to Sprague-Dawley weanling rats and beagle dogs
for 2 years. In the rat study, 60 male and 60 female control rats were fed a basal
diet and water ad lib. Three test groups of 30 male and 30 female rats each were
fed diets containing 1 .O, 3.0, or 10.0 percent Butylene Glycol for 2 years. Obser-
vations were made on body weight, food consumption, compound consump-
tion, pharmacological effects, urinalysis, and gross appearance. Erythrocyte and
leukocyte counts, packed cell volume, and hemoglobin values were determined.
At 1 year, 10 animals from each group were killed, and all survivors were killed at
the end of 2 years. Representative organs were weighed and examined micro-
scopically. These tests were negative for deleterious or toxic effects due to the in-
gestion of Butylene Glycol at any dietary concentration.‘48)
In the study using beagle dogs, a control group of 4 males and 4 females were
fed a basal diet, and 3 similar groups received diets containing 0.5, 1 .O, or 3.0
percent Butylene Glycol. Daily or weekly observations were made of feed intake,
elimination, clinical appearance, pharmacological effects, and feed and com-
pound consumption. The blood, urine, and representative organs were exam-
ined as with the rat portion of the study described above. Two animals from each
group were killed after 1 year and the remainder after 2 years. As with the rats, no
toxic effects were produced by the ingestion of Butylene Glycol at any dietary
concentration.(48)
Ethoxydiglycol was fed to rats at 1 .O g/kg per day for 2 years. The rats had
slight hepatic damage, some interstitial edema in the testes, and in 1 animal,
oxalate crystals in the kidney.(26)
Parenteral Toxicity
The subcutaneous LDso of Butylene Glycol in mice and rats was 16.5 ml/kg
and 20.1 ml/kg, respectively. (‘I) No deaths were caused by intraperitoneal injec-
tion of 1 .O g/kg Butylene Glycol into 5 mice. (4g) Butylene Glycol was evaluated
for tissue irritation using chicken pectoral muscle. Injections of 0.5 ml of the gly-
col, 1.3 cm deep into the right and left pectoral muscle of each of 6 chickens,
caused only minimal tissue irritation.(50)
Hexylene Glycol had a subcutaneous LD50 of 13 g/kg in rabbits and ro-
dents (38) The intraperitoneal LDso of Hexylene Glycol in the mouse was 4.5 ml/
kg. (51; N IOSH (35) lists the intraperitoneal LDso in mice as 1299 mglkg.
The subcutaneous LDso of Ethoxydiglycol in mice was 5500 mg/kg.(38) In rats
and rabbits, it was 3.4 ml/kg and 2.0 ml/kg, respectively.‘52’ The intraperitoneal
LDso of Ethoxydiglycol in rats was 6310. mglkg. (3*) The intravenous LDso in dogs
was 3000 mg/kg and in cats, 5000 mg/kg. (38) Other investigators report intrave-
nous LD,,s in mice, rats, and rabbits as 3.9, 2.9, and 0.9 ml/kg, respectively.(52)
The intraperitoneal LD50 of Dipropylene Glycol in rats and mice was 10 g/kg
and 4600 mg/kg, respectively. (35)The intravenous LDso in rats and dogs was 5800
mg/kg and 1 1,500 mg/kg, respectively.‘35)
Inhalation Toxicity
Rats survived an 8-hour exposure to the saturated, room temperature vapors
of Hexylene Glyc~l.(“~)
Cutaneous Toxicity
Acute Studies
The cutaneous LDso of Hexylene Glycol in rabbits and rodents was 13.2 g/
kg.(38’ A product formulation containing 5.0 percent Butylene Glycol had a cu-
taneous LD50 of >2 g/kg when tested in rabbits.(361
The cutaneous LDso of Ethoxydiglycol was 6 g/kg in ratst3*) and 10.3 g/kg in
rabbits.‘53’
A product formulation containing 7.2 percent Dipropylene Glycol produced
a cutaneous LD50 of >2 g/kg when tested in rabbits.t4”
Subchronic Studies
A product formulation containing 3 percent Butylene Glycol was applied
daily at 500 mg/kg to the clipped intact and abraded skin of each of 8 albino rab-
bits for 4 weeks. A control group of 8 rabbits remained untreated. All of the ani-
mals survived the duration of the study. Clinical observations of compound-re-
lated importance were confined to the skin, which had slight erythema with
drying and flaking. No systemic effects as evidenced by microscopic tissue exam-
ination were attributable to the test material.(54)
Skin Irritation
Primary irritation
Undiluted Butylene Glycol produced no more than minimal skin irritation
when tested under occlusion on the skin of rabbits for 24 hours’55’ or daily for 4
consecutive days.(56)
Undiluted Hexylene Glycol produced moderate irritation when 465 or 500
mg was applied to the skin of rabbits for 24 hours.‘3s) A 24-hour application of
1.84 g/kg undiluted Hexylene Glycol to the skin of rabbits caused mild edema
and erythema.(57)
Undiluted Ethoxydiglycol was a mild irritant when applied to rabbit skin (500
mg for 24 hours). (38) According to Rowe, (57) it is a nonirritant to rabbits.
Undiluted Dipropylene Glycol caused mild irritation when 500 mg was ap-
plied to rabbit skin for 24 hours.(38)
Several product formulations containing 5.0 to 21.4 percent Butylene Glycol,
1 .O to 1.6 percent Hexylene Glycol, 1 .O percent Ethoxydiglycol, or 7.2 percent
Dipropylene Glycol were tested for 24 hours under occlusion on rabbit skin
(Leberco Labs). (36,44.58-64)
Th e products produced no irritation to moderate irrita-
tion, depending upon the particular formulation tested. The degree of irritation
did not correlate with the concentration of glycol.
Cumulative Irritation
A daily dose of 0.5 ml of a paste mask product formulation containing 2 per-
cent Ethoxydiglycol was applied to the backs of 3 albino rabbits for 14 consecu-
tive days. Each treatment site was rinsed with warm tap water 30 minutes after
treatment. There was slight erythema 24 hours after the initial application that
had disappeared by 48 hours. There were no other signs of irritation.(42’
Ocular irritation
According to NIOSH, (35) 505 mg of undiluted Butylene Glycol applied to the
rabbit eye was an irritant. No irritation was observed when 0.1 ml of undiluted
Butylene Glyc01’~~’ or a 40 percent aqueous solution of Butylene Gly~ol(~~) was
instilled into 1 eye of each of 6 rabbits.
Irritation was severe when 93 mg undiluted Hexylene Glycol was instilled
into the eyes of rabbits,‘381 and Rowe’57) reported that this glycol caused cornea1
damage in a rabbit. A 25 percent aqueous solution of Hexylene Glycol caused no
ocular irritation when tested in the rabbit.(67’
Moderate toxic effects were found in the eyes of rabbits instilled with 500 mg
undiluted Ethoxydiglycol. Mild effects were caused by 125 mg.(38)
Laillier et al.(68) studied the ocular effects of Ethoxydiglycol and other chemi-
cals using the rabbit. The chemicals were applied to a series of 4 animals at appli-
cation frequencies of 1, 3, 6, 7, and 13 times over periods of 2, 4, 7, 26, and 58
hours. The chemicals were used either pure or as a 25 percent dilution in distilled
water in 0.1 ml volumes. Ocular edema was measured by the following formula:
mg dry tissue weight

x 100
mg wet tissue weight
In addition, aqueous humor and conjunctival content were assayed for effects 1
hour after Evans blue solution was injected into the rabbit’s marginal ear vein.
Ethoxydiglycol was very irritating to the rabbit’s eye in this assay system (Table 3).
Undiluted Dipropylene Glycol is an irritant in the rabbit eye in an amount of
5 10 mg. (35)
Several product formulations containing 5.0 to 21.35 percent Butylene Gly-
col, 1 .O percent Hexylene Glycol, 1 .O to 2.0 percent Ethoxydiglycol, or 7.2 per-
cent Dipropylene Glycol produced no more than minimal, transient irritation
when instilled into the eyes of rabbits.(36.42*44.69-72’Another product formulation
containing 1.6 percent Hexylene Glycol produced mild to moderate irritation in
the eyes of rabbits. (73)This formulation had also produced mild to moderate irri-
tation when applied to the skin of rabbits.
Clinical Assessment of Safety
Nutritional and Metabolic Studies

Tobin and associates(74) investigated the nutritional and human metabolic ef-
fect of Butylene Glycol. Three studies were conducted.
In the first experiment, the effect of Butylene Glycol and urea in the nutrition
of 12 men and women was studied. The volunteers went through a 2-day deple-
tion period in which nitrogen intake was 1.23 g/day. Caloric intake was evaluated
and adjusted so that during the 4- to 7-day test periods subjects consumed diets
with constant caloric content. Dietary variation was randomly distributed during
TABLE 3. Ocular Irritation in Rabbits by Measuring Tissue Edema: Ethoxydikol(68)
Conjunctivae
No. of
instillations lime* (pg Evans Blue/ Corneas Aqueous Humors
(4 Rabbits Each) (hours) (% Dry Weight) g Dry Weight) (% Dry Weight) (pg Evans Blue/ml)
Ethoxydiglycol 1 2 13.9 f 0.8+ 439 f aa+ 24.1 f 1.2+ 135.1 f 69.1+

(undiluted) 3 4 12.7 f 1.3+ 439 f 92+ 21.2 f 2.0+ 21.7 f 16.0+
6 ,7 13.1 f 0.6+ 497 f 168+ 18.6 f 1.9+ 9.0 l 4.9+
7 26 16.4 f 1~9+ 906 f 171+ 17.1 f 1.5+ 10.9 f 8.0+
13 58 15.9 * 1.0+ 988 f 283+ 15.8 f 1.4+ 29.5 f 22.9+
25% Ethoxydiglycol in 1 2 17.7 f 1.1+ 189 f 28+ 24.8 f 1.3 3.3 f 2.6+
distilled water 3 4 20.7 f 0.8 91 f 23 25.9l 1.5 0.8 f 0.25
6 7 .18.6 f 1.4 178 & 34+ 25.3 * 1.2 11.6* 8.6+
7 -26 20.8 l 0.4 177 f 32+ 25.5 f 1.5 ’ 12.8 f 11.9+
13 58 19.4 k 1.4 139 f 27 27.1 f 1.0 5.6 f 3.0+
*Time in hours over which instillations made.

+Significantly different from controls (Student’s t-test).
Mean f 95% confidence limits.
FINAL REPORT: SAFETY ASSESSMENTOF THE CLYCOLS 239
the 4 experimental periods: 1 in which the glycol (15 g) was substituted isocalori-
tally for starch in the diet, 1 in which starch without the glycol was ingested, 1 in
which urea (4 g of nitrogen/day) was added to the diet, and the fourth and final
period in which the glycol and urea were added to the diet. Butylene Glycol, as
compared to starch, caused a significant decrease of urinary nitrogen excretion.
Subjects fed urea or glycol plus urea had a significant increase in urinary excre-
tion of nitrogen. The glycol did not increase fecal nitrogen excretion, but urea
feeding did. Feeding the glycol or urea or the combination caused a less negative
nitrogen balance than did the starch feeding. The glycol caused a lowering of
blood glucose and no increase in blood ketones. The investigators concluded
that Butylene Glycol can be used as a caloric source by human beings.
The second study investigated the .effect of Butylene Glycol on endocrine
function and its influence on glucose homeostasis. Twenty-seven women volun-
teered for a 15day experiment in which one half were fed 40 g of Butylene Gly-
col per day for 5 days or a calorically equivalent quantity of sucrose for 5 days. At
the end of 5 days, the diets were switched. Mean blood glucose concentrations
and serum insulin concentrations were lower in the second and third weeks of
ingestion. The glycol had no effect on triglyceride or cholesterol concentrations.
Fasting insulin and growth hormone concentrations were somewhat increased
by the glycol.
In the third study, 10 adult male and female volunteers were fed for 12 days 6
g of nitrogen per day, starch, and vitamin and mineral supplements. The Butyl-
ene Glycol was substituted isocalorically for sucrose to provide 10 percent of the
total caloric intake. Glucose tolerance tests were performed on the sixth and
twelfth days of the test. Glucose concentrations were normal, as were free fatty
acid and growth hormone values. Serum insulin values and blood pyruvate and
lactate values were normal, and /3-hydroxybutyrate, acetoacetate and triglyceride
values were likewise normal. Butylene Glycol was nontoxic in these tests.‘74’
Skin Irritation/Sensitization
A Shelanski and Shelanski repeated insult patch test was conducted on 200
volunteers (80 male, 120 female) to assess the irritation and sensitization poten-
tials of Butylene Glycol. The compound was diluted to 50 percent in water, and
0.9 ml of the mixture was applied under occlusion to sites on the upper arm.
After 24 hours of contact, the patches were removed and the sites were graded
on a scale of 0 (no reactions) to 4+ (erythema, edema, vesicles, and extensions
beyond the site of contact). After 24 hours, the sites were reexamined. If no
changes occurred, a second patch was reapplied to the same site. This cycle was
repeated each Monday, Wednesday, and Friday. After the fifteenth application,
sites were not treated for a 2-week period, and then 24-hour occlusive patches
were applied to the same sites. Test areas were graded immediately and then at
24 and 48 hours after patch removal. The investigator reported visible skin
change in one subject after applications 4-6 and in another after applications 13-
15. No reactions were caused by the challenge patch. Butylene Glycol was a
mild fatiguing agent in 2 of 200 test subjects. With statistical extrapolation,
greater than 98 percent of the general population would not be sensitized to Bu-
tylene Glyc~l(‘~) (Table 4).
Undiluted Butylene Glycol was applied to the volar skin of the forearms or
TABLE 4. Clinical Skin Patch Tests with Butvlene Clycol and Hexylene Clycol
Concentration No. of
Test Method Material Tested Vd Subjects Results Reference
24-hour single insult Butylene Clycol 100 37 Occlusive patch; no 75

occlusive or semioc- reactions
elusive patch
39 Semiocclusive
patch; 1 subject
with mild irrita-
tion
Hexylene Clycol 100 37 Occlusive patch; 76

minimal irritation
(primary irritation
index 0.11; max,
4.0)
39 Semiocclusive
patch; minimal ir-
ritation (primary
irritation index
0.02; max, 4.0)
Shelanski and Shelanski Butylene Glycol 50 200 Mild skin irritation 19

repeated insult patch (in water) in 2 subjects; no
test (24-hour occlu- sensitization
sive patches 3 days/
week for 15 induc-
tion patches; chal-
lenge patch after 2-
week rest)
medial arms of 37 human subjects under occlusion and 39 subjects under semi-
occluded conditions for 24 hours. One subject in the semioccluded panel had
evidence of mild irritation; no other reactions were observed in either panel’75’
(Table 4). Hexylene Glycol was tested in an identical fashion, producing primary
irritation indices of 0.11 (scale 0 to 4) for the occluded patch and 0.02 for the
semioccluded patch. These scores are indicative of only minimal irritation(76)
(Table 4).
Fisher”‘) reported that cross-reactivity (sensitivity) may occur between Bu-
tylene Glycol and propylene glycol.
A number of product formulations containing 1 of the glycols at concentra-
tions of 0.016 to 21.4 percent have also been tested for skin irritation and sensiti-
zation in humans (Table 5). in single insult occlusive patch tests, products con--
taining 3.0 to 21.4 percent Butylene Glycol produced no more than minimal
irritation.‘77-so’ Several multiple insult tests were conducted on products contain-
ing a glycol in which no irritation to moderate irritation was found depending
upon the particular product tested. There was no correlation between the degree
of irritation and the concentration of glycol (Table 5). Results indicative of irrita-
tion cannot be interpreted without knowledge of the other ingredients in a for-
mulation. Of the 1087 subjects tested in skin sensitization assays (Schwartz-Peck
and Draize-Shelanski tests), there were no reactions indicative of sensitization to
any of the glycols (Table 5).
Photoreactivity
Four studies included exposure to ultraviolet light as a supplement to the
Schwartz-Peck prophetic patch tests and Draize-Shelanski repeated insult patch
tests on a nail lotion containing 5.0 percent Butylene GIy~ol(~‘) and on a shaving
preparation containing 7.2 percent Dipropylene Glyc01(~*) (Table 5). The ultravi-
olet light exposure was to a Hanovia Tanette Mark I quartz lamp at a distance of
12 inches for 1 minute. This lamp has a wavelength coverage of 240 to 370 nM,
with a peak at 365 nM. None of the subjects in the Schwartz-Peck tests had reac-
tions when a single UV exposure was made after the second insult. The Draize-
Shelanski tests included UV exposure after induction patches 1,4, 7, and 10 and
after the challenge patch; there were no reactions (Table 5).
Ocular Irritation
A drop of Butylene Glycol applied to the eyes of humans caused immediate
severe stinging similar to that induced by propylene glycol. Irrigation with water
brought rapid relief.(83)
When human subjects were exposed for 15 minutes to a vapor concentra-
tion of 50 ppm of Hexylene Glycol, ocular irritation occurred.‘57)
Inhalation Toxicity
Nasal and respiratory discomfort occurred from a concentration of 100 ppm
aerosolized Hexylene Glycol, and at 1000 ppm irritation of the eyes, nose,
throat, and respiratory tract were noted.‘“”
Industry Complaint Experience

A skin care product containing 1.6 percent Hexylene Glycol had 43 safety-re-
lated complaints in 4 years with 243 million units distributed.t9’) A shaving prepa-
ration containing 7.2 percent Dipropylene Glycol had 7 safety-related complaints
in 3 years with 2.3 million units sold; 2 of these were listed as”rash” and 5 as “irri-
tated skin.“(98)
SUMMARY
Butylene Glycol, Hexylene Glycol, Ethoxydiglycol, and Dipropylene Glycol

are viscous liquids used in the cosmetic industry as humectants, emulsifiers, plas-
ticizers, and solvents. They are added to various types of cosmetic products at
concentrations up to 50 percent. The Glycols also have many noncosmetic uses
and have been given Direct and/or Indirect Food Additive status by the FDA.
Various animal species and man metabolize Butylene Glycol and use it as a
source of calories. The results of acute, subchronic, and chronic oral toxicity
studies using a variety of animal species indicate a low order of toxicity for the
Glycols. Results of parenteral injection, inhalation, and acute and subchronic cu-
taneous toxicity studies likewise support a low order of toxicity. Butylene Glycol,
Ethoxydiglycol, and Dipropylene Glycol caused minimal to mild irritation of rab-
bit skin, whereas Hexylene Glycol was moderately irritating. The Glycols pro-
duced mild to severe ocular irritation when tested in rabbits, and Hexylene Gly-
col produced the most severe irritation. Although undiluted Hexylene Glycol
TABLE 5. Clinical Skin Patch Tests with Product Formulations Containing Glycols
No. of
TestMethod Material Jested Concentration (%i Subjects Results Reference
24-hour single insult occlusive Eye shadow 21.35 Butylene Glycol 20 Minimal irritation 79
patch
Foundation makeup 16.0 Butylene Glycol 19 Minimal irritation. Also imcluded 78
semiocclusive patch with minimal
irritation
Mascara 8.0 Butylene Glycol 20 Minimal irritation 80
Rouge 3.0 Butylene Glycol 20 No signs of irritation 77
“Soap chamber test:” 1 24-hour Personal cleanliness 0.13 Hexylene Glycol 10 Moderate irritation 84
followed by 4 daily 6-hour product (8% aqueous dilution
applications in Duhring of product containing
chambers on volar forearm 1 .6%)
Cumulative irritancy test Eye shadow 21.35 Butylene Glycol 10 Slight irritation; total composite score 85
(daily 23-hour occlusive was 70/630 max
patch for 21 days)
Paste mask 2.0 Ethoxydiglycol 12 Essentially nonirritating; total com- 86
posite score was 361630 max
Schwartz-Peck prophetic patch Nail lotion 5.0 Butylene Glycol 104 No reactions; supplemental UV ex- 81
test (open and closed 48 posure at open patch after second
hour patches, repeated after insult produced no reactions
2 weeks)
Shaving preparation 7.2 Dipropylene Glycol 101 Mild irritation with closed patch in 6 82
subjects at first exposure and in 8
subjects at second; open patches,
and supplemental UV exposure
after second insult produced no re-
actions
Draize-Shelanski repeated in- Foundation makeup 16.0 Butylene Glycol 108 Mild irritation; no sensitization 87
sult patch test (24- or 48.
hour patches 3 days/week
for 9 or 10 induction
patches; challenge patch
after 2-week rest)
Nail lotion 5.0 Butylene Clycol 49 No irritation; no sensitization. Sup- 81
plemental UV exposure after in-
duction patches 1, 4, 7, and TO
and after challenge showed no
photosensitization
Rouge 3.0 Butylene Glycol 108 Slight irritation; no sensitization 88
Eye makeup re- 1 .O Hexylene Clycol 103 Mild irritation; no sensitization 89
mover
Personal cleanliness 0.048 Hexylene Glycol 52 Mild irritation with fatiguing during 90
product (3% aqueous dilution induction: 4 reactions on challenge
of product containing at original site with 2 persisting
1 .6%) and 3 reactions on challenge at al-
ternate site with 1 persisting. Reac-
tions at challenge consistent with
induction irritation reactions. Test
sites exposed to 30 minutes natural
sunlight 24 hours after each appli-
cation
Personal cleanliness 0.016 Hexylene Glycol 52 Mild irritation; no sensitization 90
product (1% aqueous dilution
of product containing
1 .6%)
0.016 Hexylene Clycol 106 No significant irritation; no sensitira- 91
(1% aqueous dilution tion
of product containing
1 .6%)
Paste mask 2.0 Ethoxydiglycol 213 Minimal irritation; no sensitization 92
Body lotion 1 .O Ethoxydiglycol 93 Minimal irritation; no sensitization 93
Shaving preparation 7.2 Dipropylene Glycol 50 Mild irritation with probable fatigu- 82
ing; no sensitization. Supplemental
UV exposure after induction
patches 1, 4, 7, and 10 and after
challenge showed no photosensiti-
zation
Controlled use test: 4 weeks Mascara 8.0 Butylene Glycol 50 No reactions 94
Shaving preparation 7.2 Dipropylene Glycol 59 No reactions 95
Controlled use test: 2 weeks Personal cleanliness 1.6 Hexylene Clycol 80 Minimal irritation 96
product
produced severe ocular irritation, a 25 percent aqueous solution produced no

signs of irritation.
Feeding studies in man indicated that Butylene Glycol was metabolized and
nontoxic. Single insult 24hour skin patch tests on undiluted Butylene Glycol and
undiluted Hexylene Glycol showed a very low order of primary skin irritation po-
tential for these ingredients. In a repeated insult patch test, Butylene Glycol pro-
duced mild skin fatigue in 2 of 200 test subjects but no evidence of skin sensitiza-
tion. A number of product formulations containing the Glycols at concentrations
up to 21.4 percent have been tested in various human skin irritation and sensiti-
zation assays (Table 5). The degree of irritation produced depended upon the
particular formation. There was no correlation between the degree of irritation
and the concentration of the Glycol present in the formulation. There were no re-
actions indicative of skin sensitization to the Glycols in any of the 1087 subjects
tested under skin sensitization assays. Supplemental exposure to ultraviolet light
in some of the skin sensitization tests on product formulations produced no reac-
tions suggestive of phototoxicity or photosensitization. Butylene Glycol and Hex-
ylene Glycol were irritating to the human eye. And Hexylene Glycol was irri-
tating to the respiratory tract at concentrations significantly higher than those
generally found in cosmetic products.
CONCLUSION
Based on the available data, Butylene Glycol, Hexylene Glycol, Ethoxydigly-

col, and Dipropylene Glycol are safe as presently used in cosmetics.
ACKNOWLEDGMENT
Jeffrey Moore, MD, Scientific Analyst and writer, prepared the technical anal-
ysis used by the Expert Panel in developing this report.
REFERENCES
1. ESTRIN, N.F., CROSLEY, P., and HAYNES, C. (eds.). (1982). Cosmetic ingredient Dictionary, 3rd ed. Wash-
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test of product containing Butylene Glycol.* (CTFA Code 2-l 7.81).
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RAW MATERIAL SPECIFICATION

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Microbial Content Test

ACACIA SENEGAL GUM EXTRACT

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Microbial Content Test <=100 cfu/g 9184535

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Identification Match to standard JSCI methods

Specific Gravity 1.004 – 1.007 2987147

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Identification Match to standard JSCI method

Assay 99.0 % minimum JSCI method

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Identification Match to standard USP method

Assay 99.0 – 100.5 % USP method

Melting Point 125 – 128 C USP method

Items Specifications Methods

Appearance Match to standard 1476468

Odor Match to standard 2846466

Specific Gravity 0.94 - 0.98 2987147

Microbial Content Test N/A

Perfume name / code : Rose ver 1

Part II. A Specifications and test methods of raw material / ingredients

Directorate C - Public Health and Risk Assessment

SCIENTIFIC COMMITTEE ON CONSUMER PRODUCTS

the Safety Evaluation of Parabens

Adopted by the SCCP

Opinion on the safety evaluation of parabens

Opinion on the safety evaluation of parabens

In its opinion of 17 February 1999 (SCCNFP/0125/99) concerning the restrictions on materials

The SCCP was asked to answer the following questions:

Opinion on the safety evaluation of parabens

3.1. Recent Official Reports on Parabens

3.1.1. The Scientific Committee on Food (SCF), SCF 1994

Opinion on the safety evaluation of parabens

3.1.2. The European Food Safety Authority (EFSA), EFSA 2004

b) In the re-evaluation of the proliferative effects of parabens on forestomach cells in rats, it

Opinion on the safety evaluation of parabens

- NOAEL (developmental toxicity of propyl and butyl paraben) 10 mg/kg bw/day

3.2. Toxicity Profile of the parabens used in cosmetic products

3.2.1. General toxicology

3.2.2. Estrogenic effects of parabens

Opinion on the safety evaluation of parabens

3.2.3. Effects of parabens on the male reproductive system

Opinion on the safety evaluation of parabens

3.2.4. Breast cancer and the use of paraben-containing cosmetics

Methyl and ethyl paraben

Opinion on the safety evaluation of parabens

Propyl, butyl and isobutyl paraben

Butyl and isobutyl paraben

Opinion on the safety evaluation of parabens

Methyl and ethyl paraben

Propyl, isopropyl, butyl and isobutyl paraben

- full descriptions of available in vitro percutaneous absorption studies;