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Amended Safety Assessment of

Mentha piperita (Peppermint)-derived Ingredients as Used in
Cosmetics

Status: Draft Final Amended Report for Panel Review

Release Date: February 9, 2018
Panel Date: March 5-6, 2018

The 2018 Cosmetic Ingredient Review Expert Panel members are: Chair, Wilma F. Bergfeld, M.D., F.A.C.P.;
Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G.
Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR
Executive Director is Bart Heldreth, Ph.D. This report was prepared by Wilbur Johnson, Jr., M.S., Senior Scientific
Analyst.

© Cosmetic Ingredient Review

1620 L STREET, NW, SUITE 1200 ◊ WASHINGTON, DC 20036-4702 ◊ PH 202.331.0651 ◊ FAX 202.331.0088 ◊ CIRINFO@CIR-
SAFETY.ORG
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ABSTRACT: The Cosmetic Ingredient Review (CIR) Expert Panel (Panel) reviewed the safety of Mentha piperita
(peppermint)-derived ingredients; most of the ingredients function as fragrance ingredients and/or skin conditioning
agents in cosmetic products. Because final product formulations may contain multiple botanicals, each containing
similar constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching
levels that may be hazardous to consumers. Industry should continue to use good manufacturing practices to limit
impurities that could be present in botanical ingredients. The Panel reviewed data relevant to the safety of these
ingredients and concluded that Mentha Piperita (Peppermint) Oil, Leaf, and leaf-derived ingredients are safe in
cosmetics in the present practices of use and concentration, when formulated to be non-sensitizing, and that the
available data are insufficient to make a determination that the remaining 3 ingredients are safe under the intended
conditions of use in cosmetic formulations.

INTRODUCTION

The safety of four of the cosmetic ingredients named in this safety assessment has been previously
reviewed by the Panel; in 2001, the Panel issued a final report with a conclusion stating that Mentha Piperita
(Peppermint) Oil, Mentha Piperita (Peppermint) Leaf Extract, Mentha Piperita (Peppermint) Leaf, and Mentha
Piperita (Peppermint) Leaf Water are safe as used in cosmetic formulations.1 The conclusion also stated that the
concentration of pulegone, a constituent of these botanical ingredients, should not exceed 1%. In accordance with its
Procedures, CIR evaluates the conclusions of previously-issued reports every 15 years, and therefore a re-review
was initiated. The new conclusion reached in this re-review supersedes the original conclusion. According to the
web-based International Cosmetic Ingredient Dictionary and Handbook (wINCI Dictionary), most of these
ingredients are reported to function as fragrance ingredients and/or skin conditioning agents in cosmetic products.2

In addition to the four Mentha piperita (peppermint)-derived ingredients that are mentioned above, this re-
review included 6 related previously unreviewed ingredients. The complete list of ingredients included in this
assessment is:

Mentha Piperita (Peppermint) Extract Mentha Piperita (Peppermint) Leaf Extract

Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract Mentha Piperita (Peppermint) Leaf Juice
Mentha Piperita (Peppermint) Flower/Leaf/Stem Water Mentha Piperita (Peppermint) Leaf Water
Mentha Piperita (Peppermint) Leaf* Mentha Piperita (Peppermint) Meristem Cell Culture
Mentha Piperita (Peppermint) Leaf Cell Extract Mentha Piperita (Peppermint) Oil

*Previously reviewed ingredients are indicated in blue.

The cosmetic ingredient names, according to the Dictionary, are written as above, i.e., capitalized, without
italics, and unabbreviated. When referring to the plant from which these ingredients are derived, the standard
scientific practice of using italics to identify genus and species will be followed (e.g., Mentha piperita).

This safety assessment includes relevant published and unpublished data for each endpoint that is
evaluated. Published data are identified by conducting an exhaustive search of the world’s literature. A list of the
typical search engines and websites used, sources explored, and endpoints that CIR evaluates, is available on the
CIR website (https://www.cir-safety.org/supplementaldoc/preliminary-search-engines-and-websites;
https://www.cir-safety.org/supplementaldoc/cir-report-format-outline). Unpublished data are provided by the
cosmetics industry, as well as by other interested parties.

Safety test data that have been found in the published literature or those provided by the Personal Care
Products Council (Council) as unpublished data since the final report was issued, are included. Some safety test
data on menthol, menthone, and pulegone are also included in the original published final report; these data and
additional data are presented in Table 1. Considering that a limitation on pulegone is mentioned in the original
conclusion, it should be noted that a National Toxicology Program (NTP) oral carcinogenicity study with positive
results on pulegone was published in 20113 and that a 1998 publication on the absence of test substance-related
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histological cerebellar changes in Wistar rats dosed orally with pulegone is available.4 These studies are also
presented in Table 1.

Excerpts from the 2001 safety assessment on the previously reviewed ingredients are disseminated
throughout the text of this re-review document, as appropriate, and are identified by italicized text. For complete
and detailed information, please refer to the original report, which is available on the CIR website (https://www.cir-
safety.org/ingredients).

CHEMISTRY
Definition and General Characterization

Collectively, the botanical ingredients reviewed in this safety assessment are derivatives of the leaf, stem
root, and whole Mentha piperita herb. The definitions of Mentha piperita (peppermint)-derived ingredients are
stated in Table 2.2

Chemical and Physical Properties

Properties/specifications relating to Mentha piperita (peppermint)-derived ingredients are presented in
Table 3.1, 5,6,7
Method of Manufacture
Mentha Piperita (Peppermint) Oil

European and American peppermint oil is distilled with steam from the fresh, above-ground parts of the
flowering plant Mentha piperita Linne, rectified by distillation and not dementholized.1 It has been reported that the
menthone content decreases while the menthol content increases in peppermint leaves upon storage for 1 to 2
months, at 22˚C to 24˚C. However, the relative menthone to menthol proportion remained practically constant
during the total storage time.

According to one source, Mentha Piperita (Peppermint) Oil has been extracted (distilled; water solvent)
from the leaves of Mentha piperita harvested (first in July and second harvest in September) in Washington state.8

Mentha Piperita (Peppermint) Extract

According to one source, the main steps in the process of manufacturing a trade name mixture defined as
an aqueous solution containing 7.5% Mentha Piperita (Peppermint) Extract are: solubilization of Mentha piperita in
water, separation of soluble and insoluble phases, and filtration and sterilizing filtration.9

Mentha Piperita (Peppermint) Leaf Extract

The following method relates to preparation of the butylene glycol/water extract of Mentha Piperita
(Peppermint) Leaf Extract.10 Dried raw material is extracted with 50 vol% 1,3-butylene glycolic solution. After
extraction, the additional steps in the production process include: filtrate → sedimentation → filtrate → adjustment
→ packaging.

In another method, the preparation of a water/ethanol extract is described.11 Dried raw material is
extracted with 30 vol% ethanol solution. After extraction, the additional steps in the production process include:
filtrate → concentration → adjustment → sedimentation → filtrate → adjustment → packaging.

The production method described herein relates to preparation of Mentha Piperita (Peppermint) Leaf
Extract (powder form).10 According to the method of production, dried raw material is extracted with 30 vol%
ethanol solution. After extraction, the additional steps in the production process include: filtrate → concentration →
add exsiccated sodium sulfate as vehicle → drying → packaging.
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In the supercritical fluid extraction with natural carbon dioxide production method for Mentha Piperita
(Peppermint) Leaf Extract from the dried leaves of Mentha piperita, neither additives nor other technical adjuncts
are introduced during the production process.12

Mentha Piperita (Peppermint) Leaf Water

In the preparation of Mentha Piperita (Peppermint) Leaf Water, dried raw material is subjected to steam
distillation.10 After distillation, the remaining steps in the production process are: water soluble fraction obtained
→ adjustment → filtrate → packaging.
Composition and Impurities

Pulegone is found in young peppermint leaves, and is metabolized to menthol as the leaves mature. It has
also been reported that pulegone is found only in Mentha Piperita (Peppermint) Oil from young plants and in trace
amounts in “inferior” oils; pulegone is absent from “good quality” Mentha Piperita (Peppermint) Oil.1 However,
a supplier of Mentha Piperita (Peppermint) Oil reported pulegone concentrations of 1% to 4%, depending on the
origin of the oil. Published studies that have investigated the pulegone content of Mentha Piperita (Peppermint) Oil
also reported a range of <1% to 4% for Mentha Piperita (Peppermint) Oils of a North American origin.

Mentha Piperita (Peppermint) Oil

The major constituents of Mentha Piperita (Peppermint) Oil include the terpenes: (-)-menthol (30 - 55%),
(-)-menthone (14 - 32%), (+)-isomenthone (1.5 - 10%), (-)-menthyl acetate (2.8-10%), (+)-menthofuran (1.0 -
9.0%), and 1,8-cineol (3.5 - 14%).13

Certain trends were observed between oil extracted from first and second harvest leaves, and oil extracted
from fresh leaves versus dried leaves.8 When compared to the second harvest, oils from the first harvest were
generally higher in (Z)-3-hexenol, 1,8-cineol, α-pinene, β-pinene, sabinene hydrate, isomenthone, menthofuran,
pulegone, β-caryophyllene, and germacrene d, but lower in limonene, menthol, and menthone. When compared to
oils from dried leaves, oils from fresh leaves were higher in 1,8-cineol, α-pinene, limonene, isomenthone,
menthofuran, menthone, and pulegone and lower in β-caryophyllene, germacrene d, and menthol. Menthyl formate
was found in all of the Mentha piperita (peppermint) oils (from leaf extraction) that were analyzed.

Major components of the essential oil of Mentha piperita adult plants from Poland include menthone,
menthol, menthyl acetate, carvone, piperitone, 1,8-cineol, and pulegone.14

According to the United States Pharmacopeial Convention’s (USP) Food Ingredients Expert Committee,
the acceptance criteria for Mentha Piperita (Peppermint) Oil include not less than 5% total esters (calculated as
menthyl acetate) and not less than 50% menthol.7

The international standard for Mentha Piperita (Peppermint) Oil, published by the International
Organization for Standardization, contains the chromatographic profile for this ingredient that is presented in Table
4.15 Pulegone and menthofuran were among the chemicals detected. A public statement from the European
Medicines Agency on the use of herbal medicinal products containing pulegone and menthofuran indicated that
Mentha Piperita (Peppermint) Oil contains a maximum of 4% pulegone and between 1% and 9% menthofuran.16 It
was also noted that the Scientific Committee on Food (SCF) has concluded that pulegone is mainly metabolized
through pathways involving menthofuran and that these two substances show similar toxicity.

Mentha Piperita (Peppermint) Extract

Mentha Piperita (Peppermint) Extract (combined in a trade name mixture) is an aqueous solution composed
of 7.5% (maximum percentage) Mentha Piperita (Peppermint) Extract, with < 40 ppm pulegone and < 50 ppm
menthol.9 The following statement relating to composition was also provided: “Our active can be divided in sugars
(47%), mineral ashes (38%), proteins (13%), and polyphenols (2%).” Composition data on this trade name mixture
also include the following impurities: alkaloids (< 0.05 g/l; assay of alkaloids performed with Dragendorff reagent),
copper (0.23 ppm), iron (3.76 ppm), manganese (21 ppm), nickel (0.19 ppm), and zinc (3.14 ppm). In an assay of
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allergens, no allergens were detected in this aqueous solution (i.e., the concentrations were less than the sensitivity
of the method (< 1 ppm)). There also was no trace of pesticides in this aqueous solution.

The following information relating to impurities is included in a certificate of analysis for a trade name
mixture containing 2.5% Mentha Piperita (Peppermint) Extract: lead (< 10 ppm), arsenic (< 3 ppm), mercury (< 1
ppm), and pesticide residues (meets USP specification).6

Mentha Piperita (Peppermint) Leaf

The major monoterpene constituents of Mentha Piperita (Peppermint) Leaf are: (-)-limonene; 1,8-cineol;
(+)-pulegone; (-)-menthone; (+)-isomenthone; (+)-menthofuran; (-)-menthol; and (+)-neomenthol.20 Mentha
Piperita (Peppermint) Leaf also contains caffeic acid, rosmarinic acid, and the following flavonoids: apigene-,
diosmetin-, and luteolin glycosides, and free lipophile methoxylized flavones such as xanthomicrol and gardenine
D.21

The following elemental contaminants have been detected in Mentha piperita herbal tea (tea leaves)
samples (n = 3) from Serbia: manganese (111.97 mg/kg dry weight), iron (443.90 mg/kg), copper (17.15 mg/kg),
and zinc (26.86 mg/kg), molybdenum (2.695 mg/kg), cobalt (0.161 mg/kg), nickel (1.882 mg/kg), selenium (0.107
mg/kg), aluminum (554 mg/kg), and tin (3.66 mg/kg).22

It is possible that pesticide residues may be present as impurities in the leaves of Mentha piperita. In a
study in which Mentha Piperita (Peppermint) Leaves were soaked in pesticides, the dissipation rate of pesticide
residues during the drying process was said to have been satisfactory, except for the pirimiphos-ethyl pesticide,
because of its high octanol-water partition coefficient and low vapor pressure.23

Mentha Piperita (Peppermint) Leaf Extract

An analysis of Mentha Piperita (Peppermint) Leaf Extract indicated that the leaves principally contained
cinnamic acid, caffeic acid, rosmarinic acid, and various flavonoids (flavones and flavanones).17 The following
solvents were used to extract the peppermint leaves: light petroleum, dichloromethane, acetonitrile, ethyl acetate,
methanol, n-butanol, and water. Eriocitrin (383.3 ± 2.2 mg/g extract) and rosmarinic acid (381.2 ± 1.9 mg/g extract)
were the most abundant components identified within the leaves, while naringenin-7-O-glucoside (0.8 ± 0.01 mg/g
extract) was the least abundant component identified. Kynurenic acid (3.82 ± 0.46 µg/g) has also been detected in
Mentha Piperita (Peppermint) Leaf Extract.18 It should be noted that kynurenic acid is a constituent of human
synovial fluid.

Composition data provided by the Council indicate that Mentha Piperita (Peppermint) Leaf Extract
(butylene glycol/water extract) contains tannin and terpenoid (which contains 2.8 ppm pulegone) and that Mentha
Piperita (Peppermint) Leaf Extract (water/ethanol extract) contains essential oil, tannin and terpenoid.10

Data on impurities provided by the Council indicate that Mentha Piperita (Peppermint) Leaf Extract
(butylene glycol/water extract) contains not more than 10 ppm heavy metals and not more than 2 ppm arsenic.10
Mentha Piperita (Peppermint) Leaf Extract (water/ethanol extract) contains not more than 10 ppm heavy metals and
not more than 1 ppm arsenic.

The supercritical fluid extraction with natural carbon dioxide production method for Mentha piperita
(Peppermint) Leaf Extract from the dried leaves of Mentha piperita results in no solvent residues, no inorganic salts,
no heavy metals, and no reproducible microorganisms.12

According to data provided by the Council, Mentha piperita (peppermint) leaf extract powder contains not
more than 10 ppm heavy metals and not more than 2 ppm arsenic.10

A chromatographic profile for Mentha Piperita (Peppermint) Leaf Extract (CO2 extract) is presented in
Table 4.5,19
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Mentha Piperita (Peppermint) Leaf Water

Composition data provided by the Council indicate that Mentha Piperita (Peppermint) Leaf Water contains
essential oil (menthol).10 The data also included specifications that indicate that Mentha Piperita (Peppermint) Leaf
Water contains not more than 10 ppm heavy metals and not more than 1 ppm arsenic.1

USE
Cosmetic

The safety of Mentha piperita (peppermint)-derived ingredients is evaluated based on data received from
the U.S. Food and Drug Administration (FDA) and the cosmetics industry on the expected use of these ingredients
in cosmetics. Use frequencies of individual ingredients in cosmetics are collected from manufacturers and reported
by cosmetic product category in FDA’s Voluntary Cosmetic Registration Program (VCRP) database.24 Use
concentration data are submitted by the cosmetics industry in response to surveys, conducted by the Council, of
maximum reported use concentrations by product category.25

According to 2017 VCRP data, the greatest use frequency is being reported for Mentha Piperita
(Peppermint) Oil, which is being used in 827 cosmetic products (433 leave-on products + 360 rinse-off products +
34 products diluted for bath use).24 The results of a concentration of use survey conducted in 2016 indicate that
Mentha Piperita (Peppermint) Leaf Water is being used at concentrations up to 40% in leave-on products (face and
neck products [not spray]), which is the greatest use concentration that is being reported for Mentha piperita
(peppermint)-derived ingredients reviewed in this safety assessment.25 Current and historical use frequency and
concentration of use data are presented in Table 5. These data indicate that the highest maximum use concentration
of Mentha Piperita (Peppermint) Oil in cosmetics increased from 3% in 1997 to 5% in 2017. Because 1997 use
concentration data on the remaining 3 ingredients that were reviewed in the original safety assessment, Mentha
Piperita (Peppermint) Leaf Extract, Mentha Piperita (Peppermint) Leaf, and Mentha Piperita (Peppermint), were not
provided, a comparison of 1997 versus 2017 use concentration data cannot be made.

According to VCRP and Council survey data, the following Mentha piperita (peppermint)-derived
ingredients are not being used in cosmetic products: Mentha Piperita (Peppermint) Flower/Leaf/Stem Water, Mentha
Piperita (Peppermint) Flower/Leaf/Stem Extract, Mentha Piperita (Peppermint) Leaf Cell Extract, Mentha Piperita
(Peppermint) Leaf Juice, and Mentha Piperita (Peppermint) Meristem Cell Culture.

Cosmetic products containing Mentha piperita (peppermint)-derived ingredients may be applied to the skin
and hair or, incidentally, may come in contact with the eyes (at maximum use concentrations up to 0.0018 %
Mentha Piperita (Peppermint) Leaf Extract in eye lotions) and mucous membranes (at maximum use concentrations
up to 3.9% Mentha Piperita (Peppermint) Oil in bath oils, tablets, and salts). Additionally, use in lipstick products
(at maximum use concentrations up to 2.9% Mentha Piperita (Peppermint) Oil) is being reported, the application of
which may result in incidental ingestion. Products containing Mentha piperita (peppermint)-derived ingredients
may be applied as frequently as several times per day and may come in contact with the skin or hair for variable
periods following application. Daily or occasional use may extend over many years.

Mentha Piperita (Peppermint) Oil is being used in both pump hair sprays (maximum use concentrations up
to 0.02%) and aerosol hair sprays (maximum use concentrations up to 0.017%) which may result in incidental
inhalation exposure. Additionally, use of this ingredient in foot sprays at maximum use concentrations up to 0.5% is
being reported. Mentha Piperita (Peppermint) Leaf Extract is also being used in pump and aerosol hair sprays, but
at lower maximum use concentrations, and in face and neck/body and hand spray products at maximum use
concentrations up to 0.001%. Mentha Piperita (Peppermint) Extract is being used in face and neck sprays at a
highest maximum use concentration of 1.3%. In practice, 95% to 99% of the droplets/particles released from
cosmetic sprays have aerodynamic equivalent diameters > 10 µm, with propellant sprays yielding a greater fraction
of droplets/particles below 10 µm, compared with pump sprays.26,27,28,29 Therefore, most droplets/particles
incidentally inhaled from cosmetic sprays would be deposited in the nasopharyngeal and bronchial regions and
would not be respirable (i.e., they would not enter the lungs) to any appreciable amount.26,27
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Mentha Piperita (Peppermint) Oil is being used in foot powders at maximum use concentrations up to
1%, and Mentha Piperita (Peppermint) Leaf Extract is being used in face powders at maximum use concentrations
up to 0.0018%. Conservative estimates of inhalation exposures to respirable particles during the use of loose
powder cosmetic products are 400-fold to 1000-fold less than protective regulatory and guidance limits for inert
airborne respirable particles in the workplace.30,31,32

Noncosmetic

Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Extract,

Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract,
and Mentha Piperita (Peppermint) Leaf Juice

Mentha Piperita (Peppermint) Oil is a generally recognized as safe (GRAS) ingredient according to the US
FDA for use in dietary supplements.1 It is described as a naturally occurring carminative that relaxes
gastrointestinal smooth muscle. A final ruling by the FDA labeled Mentha Piperita (Peppermint) Oil as safe and
effective as an antitussive (topical/inhalant). Final rulings cautioned that Mentha Piperita (Peppermint) Oil is not
safe and effective for use as an expectorant in either topical/inhalant or lozenge form, or for use as a nasal
decongestant, mouthwash, or digestive aid.

Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Extract, Mentha Piperita (Peppermint)
Flower/Leaf/Stem Extract, and Mentha Piperita (Peppermint) Leaf Juice are generally recognized as safe (GRAS) by
the US FDA for use in food for human consumption (21CFR182.20). Mentha Piperita (Peppermint) Oil is an
inactive ingredient in drug products that have been approved by the U.S. FDA.33 A number of active ingredients,
Mentha Piperita (Peppermint) Oil included, have been present in over-the-counter (OTC) drug products for various
uses.34 However, the FDA has determined that, based on evidence currently available, there are inadequate data to
establish general recognition of the safety and effectiveness of Mentha Piperita (Peppermint) Oil as an active
ingredient in the following drug products: nasal decongestant drug products, digestive aid drug products, insect bite
and sting drug products, and astringent drug products.

Mentha Piperita (Peppermint) Oil is on the U.S. Environmental Protection Agency (EPA) list of active
ingredients eligible for minimum risk pesticide products.35

Mentha Piperita (Peppermint Oil)

According to the European Medicines Agency Committee on Herbal Medicinal Products (HMPC)
community herbal monograph on Mentha x piperita L., aetheroleum (i.e., peppermint oil; aetheroleum is a term used
to describe a preparation made from the above-ground parts of a plant), this herbal medicine is administered orally
for the symptomatic relief of minor spasms of the gastrointestinal tract, flatulence, and abdominal pain,
especially in patients with irritable bowel syndrome.36 It is also an herbal medicine that is administered
cutaneously for the symptomatic relief of mild tension-type headache. These uses have been identified as well-
established uses by the HMPC. The highest recommended daily dose in the European Union is 1.2 ml peppermint
oil (i.e. 1,080 mg peppermint oil, which contains maximum 140 mg pulegone + menthofuran). For a 60 kg person,
this would correspond to a daily intake of 2.3 mg/kg body weight.

TOXICOKINETIC STUDIES

Dermal Penetration

Mentha Piperita (Peppermint) Oil

Eserine in a Mentha Piperita (Peppermint) Oil vehicle was applied to a 2.2 cm2 shaved area on the
abdomen of mice. The absorption rate for Mentha Piperita (Peppermint) Oil was measured as the latent period
between application and appearance of eserine-induced signs.1 Mentha Piperita (Peppermint) Oil had a latent
period of 58 minutes.
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Penetration Enhancement

Mentha Piperita (Peppermint) Leaf Extract

The skin penetration enhancement potential of Mentha Piperita (Peppermint) Leaf Extract (aqueous ethanol
extract) was evaluated using dorsal porcine skin (dermatomed to thickness of 500 µm).37 A square section of skin
was cut to provide a dose area of 1 cm2 and placed in a flow-through diffusion cell. [14C]-Caffeine (hydrophilic) or
[14C]-salicylic acid (hydrophobic) was applied topically with 10% Mentha Piperita (Peppermint) Leaf Extract to
porcine skin. The receptor fluid for the diffusion cell was “a Krebs-Ringer bicarbonate buffer spiked with dextrose
and BSA (acronym not defined)”. Ethanol alone served as the control. When compared to [14C]-caffeine in the
presence of ethanol (control), the dermal absorption of [14C]-caffeine was significantly greater (p > 0.05; flux and
permeability of caffeine increased by over 3-fold) in the presence of Mentha Piperita (Peppermint) Leaf Extract.
However, this was not true for [14C]-salicylic acid.
Penetration Inhibition

Mentha Piperita (Peppermint) Oil

Mentha Piperita (Peppermint) Oil and ring-UL-[14C]benzoic acid were applied to full-thickness human skin
(breast or abdominal) samples in a static diffusion cell.38 The receptor fluid for the diffusion cell was “0.9% sodium
chloride and 1% Tween in water”. As the concentration of Mentha Piperita (Peppermint) Oil increased from zero to
5% in the donor phase, the maximal flux of benzoic acid decreased. The differences were significant at 1.0% and
5.0% Mentha Piperita (Peppermint) Oil, where the maximal fluxes were reduced to 81% and 52% of the control,
respectively.
Absorption, Distribution, Metabolism, and Excretion

Oral
Mentha Piperita (Peppermint) Oil

The rate of Mentha Piperita (Peppermint) Oil absorption and excretion following oral administration was
determined by measuring urinary menthol glucuronide.1 Four male volunteers ingested 180 mg of an enteric-coated
Mentha Piperita (Peppermint) Oil-coated capsule following a 16-h fast. Menthol was liberated from its glucuronide
metabolite by treating the urine with β-D-glucuronidase. It was estimated that between 37 and 116 mg of menthol
corresponding to an average 40% recovery of the administered menthol dose was excreted by each panelist within
14 h.

Mentha Piperita (Peppermint) Oil is relatively rapidly absorbed after oral administration and eliminated
mainly via the bile.13 The major biliary metabolite is menthol glucuronide, which undergoes enterohepatic
circulation. The urinary metabolites result from hydroxylation at the C-7 methyl group at C-8 and C-9 of the
isopropyl moiety, forming a series of mono- and dihydroxymenthols and carboxylic acids, some of which are
excreted, in part, as glucuronic acid conjugates. Studies with tritiated L-menthol in rats indicated approximately
equal excretion in the feces and urine. The main metabolite identified was menthol-glucuronide. Additional
metabolites are mono- or di-hydroxylated menthol derivatives.

TOXICOLOGICAL STUDIES

Acute Toxicity Studies

Oral
Mentha Piperita (Peppermint) Oil
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Mentha Piperita (Peppermint) Oil had a 24-h oral LD50 of 4441 mg/kg in fasted Wistar rats; the 48-h LD50
was 2426 mg/kg.1 In a study involving fasted mice, an LD50 of 2410 mg/kg was reported for Mentha Piperita
(Peppermint) Oil diluted in olive oil.

Short-Term Toxicity Studies

Oral
Mentha Piperita (Peppermint) Oil

In 3 of 4 short-term oral toxicity studies (28-day or 5-week studies) involving 20 to 28 rats per group, brain
lesions (specifically, cyst-like spaces in the cerebellum) were observed at Mentha Piperita (Peppermint) Oil doses
up to 100 mg/kg/day.1 In the remaining study (12 rats per group), these lesions were not observed in rats dosed with
Mentha Piperita (Peppermint) Oil at doses of 20, 150, or 500 mg/kg/day for 5 weeks.
Subchronic Toxicity Studies

Oral
Mentha Piperita (Peppermint) Oil

Groups of 28 Wistar rats were given oral doses of 10, 40, and 100 mg/kg Mentha Piperita (Peppermint) Oil
(diluted with soybean oil) daily for 90 days.1 All hematological and biochemical parameters were within normal
range, and there were no significant differences in absolute and relative organ weights. Brain lesions (specifically,
cyst-like spaces in the cerebellum) were observed in all dose groups, but these results were classified as significant
only for animals of the 100 mg/kg/day dose group. No other lesions of encephalopathy were observed. Nephropathy
(hyaline droplet formation) was observed only in male rats of the 100 mg/kg/day dose group, and there was no
evidence of epithelial degeneration. The no-observed-adverse-effect level (NOAEL) for Mentha Piperita
(Peppermint) Oil was 40 mg/kg/day in this study.

GENOTOXICITY STUDIES
In Vitro

Mentha Piperita (Peppermint) Oil

The mutagenic potential of Mentha Piperita (Peppermint) Oil was investigated using the
Salmonella/mammalian microsome test.1 The following Salmonella typhimurium strains were used: TA1535,
TA100, TA1537, and TA98. The sample tested contained 38.1% menthol, 33.7% menthone, and 1.7% pulegone; the
remaining components were not identified. Mentha Piperita (Peppermint) Oil, tested at doses of 6.4, 32, and 160
µg/plate, produced the same number of revertants as the negative control. Toxicity was noted at the next (and
maximum) dose of 800 µg/plate. Metabolic activation appeared to have made the oil less toxic to the bacteria.
Mentha Piperita (Peppermint) Oil was not genotoxic.

In an in vitro chromosomal aberration test using a Chinese hamster fibroblast cell line, Mentha Piperita
(Peppermint) Oil, at a maximum concentration of 0.25 mg/ml (in ethanol), produced polyploidism in 3% of the cells
and structural aberrations in 7% of the cells at 48 h after treatment. The results were considered equivocal, as
scores of either ≥ 10% or ≤ 4.9% were necessary for classification as either positive or negative, respectively. The
results for Mentha Piperita (Peppermint) Oil (150 µg/ml) were negative in a mouse lymphoma L5178Y TK +/- cell
mutagenesis assay. Results were also negative for this ingredient (at 155 µg) in an unscheduled DNA synthesis
assay using rat hepatocytes.1

The genotoxicity of Mentha Piperita (Peppermint) Oil was evaluated in a chromosome aberration test using
human peripheral blood lymphocytes.39 Lymphocyte cultures were incubated for 24 h with test substance
concentrations up to 0.30 µl/ml. When chromosome aberrations (chromatid breaks, chromatid exchanges,
chromosome breaks, and chromosome exchanges) were scored, not less than 100 metaphases per culture were
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analyzed. Mentha Piperita (Peppermint) Oil was the most clastogenic at a concentration of 0.20 µl/ml (8-fold
increase over acetone solvent control); the number of aberrant cells decreased at higher concentrations. The authors
noted that the dose-response curve for Mentha Piperita (Peppermint) Oil was complicated, with a clear peak
response at a concentration of 0.20 µl/ml.

Mentha Piperita (Peppermint) Oil was tested at concentrations up to 0.30 µl/ml in the sister chromatid
exchange (SCE) test involving human lymphocytes.39 The test conditions were essentially the same as those in the
preceding chromosome aberration test, with the exception that 5-bromo-2'-deoxyuridine was added (10 µg/ml) to
cultures initially. To determine the replicative index, 200 cells were scored. Mentha Piperita (Peppermint) Oil
induced SCEs in a dose-independent manner. The authors noted that, seemingly, the saturation of SCE-inducing
capacity occurred at high concentrations of Mentha Piperita (Peppermint) Oil. Results also indicated that Mentha
Piperita (Peppermint) Oil inhibited cell replicative kinetics, some signs of which were observed at a concentration of
0.15 µl/ml. At concentrations ≥ 0.20 µl/ml, statistically significant inhibition of cell replicative kinetics was evident.

Mentha Piperita (Peppermint) Extract

The genotoxicity of a trade name mixture containing 2.5% Mentha Piperita (Peppermint) Extract was
evaluated in the Ames test using the following Salmonella typhimurium strains, with and without metabolic
activation: TA97a, TA98, TA100, TA102, and TA1535.40 The test substance was diluted with sterile distilled water
to a concentration of 10% (effective concentration of extract = 0.25%) prior to testing each strain. Sterile deionized
water served as the solvent control and positive controls (not stated) were also used. The test substance was not
cytotoxic to the test system and was not genotoxic to any of the strains tested, either with or without metabolic
activation. The bacterial strains tested were sensitive to the positive control mutagens and had a spontaneous
reversion rate that was well within the accepted values for each strain.

ANTIGENOTOXICITY STUDIES

Mentha Piperita (Peppermint) Leaf Extract

Oral pretreatment with Mentha Piperita (Peppermint) Leaf Extract (aqueous extract) (1 g/kg/day for 3
consecutive days) before exposure to gamma radiation was found to be effective in protecting against chromosomal
damage in the bone marrow of Swiss albino mice (number tested not stated).41 The exposure of mice to 8 gray (Gy)
ionizing radiation (1 Gy is equivalent to the absorption of one joule of radiation energy per kilogram of matter) only
resulted in chromosomal aberrations in the form of chromatid breaks, chromosome breaks, centric rings, dicentrics,
exchanges, and acentric fragments. In mice pretreated with Mentha Piperita (Peppermint) Leaf Extract, there was a
significant decrease in the frequency of aberrant cells when compared to the irradiated control. A significant
increase in the percentage of chromatid breaks, chromosome breaks, centric rings, dicentrics, exchanges, acentric
fragments, total aberrations, and aberrations/damaged cell was observed at 12 h post-irradiation necropsy time in
control animals. However, a significant decrease in the percentage of aberrations of this type was observed in mice
pretreated with Mentha Piperita (Peppermint) Leaf Extract.

The modulatory effects of Mentha Piperita (Peppermint) Leaf Extract (aqueous extract) on genotoxicity and
lung tumor incidence were evaluated using 4 groups of 30 to 76 Swiss albino mice.42 Beginning at 3 weeks of age
(weaning), the mice received a single subcutaneous injection of benzo[a]pyrene and were then dosed orally (by
gavage) with either water (group of 53 mice) or Mentha Piperita (Peppermint) Leaf Extract (1 g/kg; group of 76
mice). The remaining 2 groups of mice in the study were identified as no benzo(a)pyrene or Mentha Piperita
(Peppermint) Leaf Extract dosing (30 mice) and Mentha Piperita (Peppermint) Leaf Extract alone (30 mice). When
compared to mice in the benzo(a)pyrene only group, Mentha Piperita (Peppermint) Leaf Extract reduced the
frequency of chromosomal aberrations and micronuclei in bone marrow cells. Mentha Piperita (Peppermint) Leaf
Extract had an antigenotoxic effect in this study. Results relating to the modulatory effect of Mentha Piperita
(Peppermint) Leaf Extract on lung tumor formation are included in the Anticarcinogenicity section of the report text.
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CARCINOGENICITY STUDIES

Oral
Mentha Piperita (Peppermint) Oil

In a carcinogenicity study of toothpaste and its components, groups of 52 male pathogen-free CFLP (ICI-
redefined) mice were dosed by gavage with 4 or 16 mg Mentha Piperita (Peppermint) Oil/kg/day, 6 days per week
for 80 weeks. Treatment was followed by a 16- to 24-week observation period. An untreated group of 52 male mice
and a vehicle control group of 260 male mice that received the toothpaste base (which did not contain chloroform,
eucalyptol, or Mentha Piperita (Peppermint) Oil) were maintained as controls. At least one neoplasm at any site
was observed in 73%, 69%, 65%, and 71% of mice of the low-dose, high-dose, untreated control, and vehicle
control groups, respectively. The incidence of neoplasms of the lungs and kidneys was comparable among mice of
the treated and nontreated groups. Hepatic cell tumor incidence for Mentha Piperita (Peppermint) Oil-dosed mice
(25%) was comparable to the incidence for mice of the vehicle control group (27%); the incidence for the untreated
group was 19%. Malignant lymphoma was found in 25%, 21%, 10%, and 14% of mice of the low-dose, high-dose,
untreated, and vehicle control groups, respectively. The researchers did not discuss whether the differences in
tumor incidence were significant.1

ANTICARCINOGENICITY STUDIES

Mentha Piperita (Peppermint) Leaf Extract

The modulatory effects of Mentha Piperita (Peppermint) Leaf Extract (aqueous extract) on genotoxicity and
lung tumor incidence were evaluated using 4 groups of 30 to 76 Swiss albino mice.42 The dosing procedure
(including groups tested) was identical to that stated for Swiss mice in the second study of the Antigenotoxicity
section in this report. The mice were killed at 9 weeks of age and evaluated for lung tumor incidence. Dosing with
Mentha Piperita (Peppermint) Leaf Extract caused a significant reduction in the number of lung adenomas from an
incidence of 67.92% in mice given benzo[a]pyrene only to 26.31%, which amounted to 61.26% inhibition. Tumor
multiplicity was 1.22 in the benzo[a]pyrene only group and 1.15 in the benzo[a]pyrene + Mentha Piperita
(Peppermint) Leaf Extract group. Mentha Piperita (Peppermint) Leaf Extract had an inhibitory effect on lung tumor
formation in this study. Results relating to the modulation of genotoxicity are included in the Antigenotoxicity
section of the report text.

The anticancer potential of Mentha Piperita (Peppermint) Leaf Extract (double-distilled; water extract) was
studied using Swiss albino mice (number not stated).43 Two stage mouse skin carcinogenesis was initiated by
7,12-dimethyl-benz[a]anthracene (DMBA). Two weeks later, croton oil (promoter) was applied 3 times per week
for 14 weeks. The mice were dosed orally with Mentha Piperita (Peppermint) Leaf Extract (800 mg/kg/day) for the
same period. At the end of the dosing period, average latent period, tumor incidence, size, burden, weight and
cumulative number of papillomas were assessed. Dosing with Mentha Piperita (Peppermint) Leaf Extract caused
inhibition of skin papilloma formation induced by DMBA and the application of croton oil, in terms of a significant
decrease in the cumulative number of papillomas, tumor burden, and tumor incidence. In the control group, the
tumor incidence was 100 percent. However, after dosing with the test substance for 15 days, the tumor incidence
was reduced to 64%. There was a significant increase in the latency period for the appearance of papillomas in test
animals (11 weeks in control group; 13 weeks in test group).

The possible molecular mechanisms underlying the cytotoxicity and anticarcinogenic potential of Mentha
Piperita (Peppermint) Leaf Extract (petroleum ether, benzene, chloroform, ethyl acetate, methanol, or water extract)
on 6 human cancer (HeLa, MCF-7, Jurkat, T24, HT-29, MIAPaCa-2) and normal (IMR-90, HEK-293) cell lines
were evaluated.44 In the human cancer cell lines tested with doses of 1 µg/ml, 10 µg/ml, and 100 µg/ml for 6 h, the
number of apoptotic cells was incremental with an increase in the dose of Mentha piperita extracts. However, of all
the extracts tested, the chloroform and ethyl acetate extracts resulted in a significantly higher apoptotic index after 6
hours, and the results were dose-dependent. When compared to the cancer cell lines, no significant changes were
observed in normal cells. Similarly, of all of the extracts tested, the chloroform and ethyl acetate extracts of Mentha
piperita had significant dose- and time-dependent anticarcinogenic activity, leading to G1 cell cycle arrest and
mitochondrial-mediated apoptosis, perturbation of oxidative balance, upregulation of Bax gene, elevated expression
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of p53 and p21 in the treated cells, and acquisition of senescence phenotype, while inducing pro-inflammatory
cytokines response.

A study was performed to evaluate the antitumor activity of Mentha Piperita (Peppermint) Leaf Extract
(methanol extract), using SW-480 human colon adenocarcinoma cells in a relevant cell anti-proliferation assay.45
Statistically significant (α = 0.05) growth inhibition was observed at a concentration of 31µg/ml. An IC50
(concentration required for 50% inhibition, µg/ml) of 92.3 µg/ml was reported for Mentha Piperita (Peppermint)
Leaf Extract.

OTHER RELEVANT STUDIES

Cytotoxicity

Mentha Piperita (Peppermint) Oil

The cytotoxicity of Mentha Piperita (Peppermint) Oil was evaluated in the (3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide (MTT) assay using 2 human cancer cell lines, MCF-7 and LNCaP.46 Mentha
Piperita (Peppermint) Oil from plants that were harvested during the summer and winter was tested. The following
IC50 values (µg/ml) were reported: MCF-7 cell line (75.2 ± 2.9 [summer]; 80.8 ± 3.2 [winter]) and LNCaP cell line
(90.4 ± 3.7 [summer]; 95.7 ± 4.5 [winter]). IC50 values in the 10 to 100 µg/ml range represented a potentially toxic
chemical, and IC50 values < 10µg/ml represented a potentially very toxic chemical.

In another study, essential oil was extracted from the leaves of Mentha piperita.47 The oil essential oil was
found to be cytotoxic in the following 4 human cancer cell lines: human lung carcinoma SPC-A1 cells (IC50 = 10.89
µg/ml), human leukemia K562 cells (IC50 = 16.16 µg/ml) and human gastric cancer SGC-7901 cells (IC50 = 38.76
µg/ml). The essential oil was inactive against human hepatocellular carcinoma BEL-7402 cells.

Mentha piperita

The inhibitory effect of polysaccharides extracted from Mentha piperita on A549 non-small cell lung
adenocarcinoma cells was investigated using the MTT assay.48 The results indicated that polysaccharides extracted
from Mentha piperita had a moderate toxic effect on the A549 cell line (IC50 = 879.52 ± 22.55 μg/ml). The growth
of A549 cells was inhibited by Mentha piperita in a dose-dependent manner. The inhibitory rate was 54.54% ±
1.38% at the highest concentration tested (1 mg/ml).

Hepatotoxicity

Mentha Piperita (Peppermint) Leaf Extract

Mentha Piperita (Peppermint) Leaf Extract (methanol extract) and other botanical extracts were tested on
both human (HepG2/C3A) and rat (MH1C1) hepatoma cells, using a battery of toxicity endpoints.49 The extract was
dissolved in dimethyl sulfoxide (DMSO) and then diluted in culture medium to a final concentration of 1000 µg/ml.
The following 8 endpoints covering a variety of biological activities relevant to hepatotoxicity were used for
hepatotoxicity evaluation: oxidative stress, mitochondrial membrane permeability, cellular neutral and polar lipid
accumulation, CYP1A, 2B, 3A activities, albumin excretion, and total DNA content. Cluster analysis was used to
group the phenolics into 4 clusters for each cell type. Two of the clusters were cluster 1 (compounds clustering with
the solvent control (DMSO) and cluster 2 (compounds with reported in vivo liver toxicity). Overall and individual
liver activity of the phenolics on both human and rat hepatoma cell lines were compared. For HepG2/C3A cells,
100% of the observations for Mentha Piperita (Peppermint) Leaf Extract and thyme extract, 92% for cinnamon
extract, and 89% for juniper berry extract were assigned to cluster 1 (control group). For rat MH1C1 cells, 100% of
the juniper berry extract and Mentha Piperita (Peppermint) Leaf Extract observations were assigned to cluster 1.
The authors noted that because there are currently no reports of liver toxicity associated with peppermint, Mentha
Piperita (Peppermint) Leaf Extract is useful as a negative control.
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Nephrotoxicity

Mentha Piperita (Peppermint) Leaf Extract

The effects of Mentha Piperita (Peppermint) Leaf Extract (“Mentha piperita tea” (i.e., aqueous extract)) on
rat kidney tissue were evaluated. The tea (prepared daily) was made by pouring 250 ml of boiling water over one
heaped teaspoon (5 g) of the dried leaves of Mentha piperita L (grown in Turkey) and steeping for 5 to 10 minutes.
Groups of 12 male Wistar albino rats were used. Test animals received Mentha piperita tea (20 g/l) in drinking
water for 30 days. Control rats were given commercial drinking water during the study. The following
histopathological changes, described as slight, were reported for the group dosed with Mentha piperita tea: hydropic
degeneration of tubular epithelial cells, epithelial cells with pyknotic nuclei and eosinophilic cytoplasm, tubular
dilatation and enlargements in Bowman capsules. In conclusion, the results indicate that Mentha piperita is not
nephrotoxic to rats.50

Effect on Histamine Release

Mentha Piperita (Peppermint) Leaf Extract

The 50% ethanol extract of peppermint leaves and stems significantly inhibited histamine release from rat
peritoneal mast cells that was induced by compound 48/80 (polymer produced by the condensation of
N-methyl-p-methoxyphenethylamine with formaldehyde) in vitro.51

Mentha Piperita

In a study involving human basophil cell suspensions, obtained from workers who were exposed to an
additive containing penicillin, the cell suspensions were incubated with 10-1 to 10-3 mg/ml Peppermint (dry aroma).
A dose-dependent increase in histamine release was noted, and it was concluded that this release was due to
nonimmunological mechanisms.1
Immune System Effects

Mentha Piperita (Peppermint) Oil

The results of a host-resistance assay involving groups of 20 mice that had been dosed orally with Mentha
Piperita (Peppermint) Oil (up to 1250 mg/kg/day for 5 days) suggested immunosuppression and/or increased
susceptibility to bacterial-induced mortality. The results of a plaque-forming assay involving groups of 10 mice that
received the same oral doses were negative.1

Effect on Hair Growth

In a study involving C57BL/6 mice, the data suggest that 3% Mentha Piperita (Peppermint) Oil (diluted in
jojoba oil) facilitates hair growth by promoting the conservation of vascularization of hair dermal papilla, which
may contribute to the induction of early anagen stage.52

DERMAL IRRITATION AND SENSITZATION STUDIES

Dermal irritation and sensitization studies are summarized in Table 6.

Irritation
In Vitro
Mentha Piperita (Peppermint) Leaf Extract
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The skin irritation potential of Mentha Piperita (Peppermint) Leaf Extract (water/ethanol extract) at
concentrations of 10% and 100% was evaluated using the in vitro reconstructed human epidermis test method, and
results were negative.

Animal
Mentha Piperita (Peppermint) Oil

Hairless sites on 5 white rabbits were injected intradermally with 0.05 ml Mentha Piperita (Peppermint)
Oil. Gross examinations were performed at 24 h and 48 h, at 1 and 2 weeks, and, in some cases, at 1 month after
dosing. Dosing was repeated between 5 and 10 times. At microscopic examination of skin samples, moderate
reactions characterized by polymorphonuclear leucocytes, lymphocytes, and plasma cells (without necrosis) were
observed in 3 rabbits. Severe reactions, which were marked by the above as well as necrosis, were observed in the
other 2 rabbits.1

Mentha Piperita (Peppermint) Extract

The skin irritation potential of a trade name mixture containing 7.5% Mentha Piperita (Peppermint) Extract
was evaluated using 3 rabbits (strain not stated).9 No cutaneous reactions were observed, and the authors concluded
that the mixture was a non-irritant.

Human
Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Leaf Extract,
and Mentha Piperita (Peppermint) Leaf Water

The skin irritation potential of a lipstick containing 0.2961% Mentha Piperita (Peppermint) Leaf Extract
was evaluated in a 48-h occlusive patch test using the following group of 50 subjects: normal (25), with eczema (4
subjects), with allergy (4 subjects) and with sensitive skin (17 subjects). Results were classified as negative.53
Slight erythema was observed in 1 of 50 subjects after repeated applications of a cleaning gel containing 50%
Mentha Piperita (Peppermint) Leaf Water in a product use study.54 Mild and moderate erythema were observed in
12 and 6 subjects, respectively, patch tested with 50% Mentha Piperita (Peppermint) Leaf Water (10% aqueous
solution dilution; effective concentration = 5% Mentha Piperita (Peppermint) Leaf Water).55 In one of the skin
sensitization studies on 20% Mentha Piperita (Peppermint) Oil that is summarized in the following section, it was
reported that there was no evidence of skin irritation in the 104 subjects tested.56

Sensitization
Animal
Mentha Piperita (Peppermint) Extract

The skin sensitization potential of a trade name material containing 7.5% Mentha Piperita (Peppermint)
Extract was evaluated in the maximization test using 10 albino guinea pigs.9 No macroscopic cutaneous reactions
attributable to allergy were associated with application of the trade name material. There also were no cutaneous
intolerance reactions in animals of the negative control group (further details not provided).

Human
Mentha Piperita (Peppermint) Oil

In the maximization test, 25 healthy male panelists received five 48-h occlusive induction patch (containing
8% Mentha Piperita (Peppermint) Oil) applications. Pre-treatment was for 24 h with an occlusive patch containing
5% sodium lauryl sulfate (SLS) prior to each exposure. After a 10-day non-treatment period, the subjects were
challenged on the back with a 48-h patch (also preceded by SLS treatment). No evidence of sensitization was
found.1
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Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Extract, Mentha Piperita (Peppermint)
Leaf Extract, and Mentha Piperita (Peppermint) Leaf Water

In a human repeated insult patch test (HRIPT) on 20% Mentha Piperita (Peppermint) Oil involving 104
subjects, results were negative for skin irritation and sensitization. 56 Skin sensitization also was not observed in
another HRIPT on 20% Mentha Piperita (Peppermint) Oil involving 101 subjects.57 An HRIPT on a trade name
mixture containing 2.5% Mentha Piperita (Peppermint) Extract was performed using 52 male and female subjects,
and results were negative for dermal irritation and allergic contact sensitization.58 Results were also negative in an
HRIPT evaluating the cumulative irritation and/or allergic contact sensitization potential of a cosmetic product
containing 0.00554% Mentha Piperita (Peppermint) Extract in 51 subjects.59 In a maximization test on a cosmetic
product containing 0.00554% Mentha Piperita (Peppermint) Extract involving 26 subjects, there was no evidence of
contact allergy.60 HRIPT results for undiluted Mentha Piperita (Peppermint) Leaf Extract (water/ethanol extract) in
52 subjects were also negative.11 A face cream containing 20% Mentha Piperita (Peppermint) Leaf Water did not
induce cumulative skin irritation or sensitization in an HRIPT involving 107 subjects.61

Photosensitization/Phototoxicity

Animal
Mentha Piperita (Peppermint) Oil

Undiluted Mentha Piperita (Peppermint) Oil was applied to the backs of 6 Skh:hairless mice. Thirty
minutes later, the mice were irradiated for either 1 h with light from a fluorescent blacklight at an integrated UVA
of 3 W/m2, or for 40 minutes with light from a Xenon lamp at a weighted erythema energy of 0.1667 W/m2. The mice
were examined at 4 h, 24 h, 48 h, 72 h, and 96 h after radiation treatment. No effects were noted. In a second
experiment, using 2 miniature swine and following the same protocol, no effect was produced by 100% Mentha
Piperita (Peppermint) Oil.1

OCULAR IRRITATION STUDIES

In Vitro
Mentha Piperita (Peppermint) Extract

The ocular irritation potential of a trade name mixture containing 2.5% Mentha Piperita (Peppermint)
Extract was evaluated using an in vitro toxicity testing system consisting of normal, human-derived epidermal
keratinocytes.63 The cells had been cultured to form a stratified squamous epithelium that is similar to that found in
the cornea. The procedure utilized a tetrazolium salt (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium
bromide (MTT)) that is reduced by succinate dehydrogenase (in viable mitochondria of viable cells) to a formazan
derivative. The amount of MTT that is reduced by a culture is proportional to the number of viable cells. The trade
name mixture, at a concentration of 10% in corn oil (effective concentration of extract = 0.25%) and a volume of
100 µl, was added to cell cultures; the incubation periods were 1 h, 4 h, and 24 h. Corn oil served as the negative
control. An ET50 (time of exposure needed for a test material to reduce the viability of treated tissues to 50% of
control tissues) was calculated. Values for % viability were: 108% (at 1 h), 100% (at 4 h), and 34% (at 24 h).
Results indicated that the trade name mixture at a concentration of 10% (ET50 = 15.5 h (non-irritating, minimal)) had
an ocular irritation potential that was somewhat less that sodium dodecyl sulfate at a concentration of 0.3% (ET50 =
740 minutes (12.3 h)).

Animal
Mentha Piperita (Peppermint) Extract

A trade name mixture containing 7.5% Mentha Piperita (Peppermint) Extract was instilled (0.1 ml) into 1
eye of each of 3 New Zealand rabbits.9 Slight conjunctival redness was observed in 2 animals and lacrimation was
observed in 1 animal. The trade name mixture was classified as a slight ocular irritant.
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Human
Mentha Piperita (Peppermint) Leaf Water

The ocular irritation potential of a cleansing gel containing 50% Mentha Piperita (Peppermint) Leaf Water
was studied using 50 subjects.54 The subjects applied the product twice per day for 4 weeks, and were instructed to
record any signs felt or observed during product use. Product use did not cause any signs of ocular or palpebral
irritation.

CLINICAL STUDIES
Multicenter Studies

Mentha Piperita (Peppermint) Oil

Data from multicenter studies evaluating the skin irritation/sensitization potential of Mentha Piperita
(Peppermint) Oil in patients are summarized in Table 6.62,64

A multicenter study involving 13,398 patients was performed by the US/Canadian North American Contact
Dermatitis Group (NACDG), whereby 71 patients were patch tested with Mentha Piperita (Peppermint) Oil (2% in
petrolatum). A positive reaction prevalence rate of 0.53% was reported for this ingredient.64 In another
multicenter study, neither irritant nor allergic reactions were observed in 73 patients patch tested with Mentha
Piperita (Peppermint) Oil according to International Contact Dermatitis Research Group (ICDRG) patch test
procedures.62
Case Reports

Case reports are summarized in Table 6.

Positive patch test reactions to Mentha Piperita (Peppermint) Oil and Mentha Piperita were reported in 8 of
9 case reports on patients with diseased skin.65,66,67,68,69,70,71 Though positive patch test results were reported in one of
the studies on Mentha Piperita (Peppermint) Oil, prick test results in that study were negative. In patients without
skin disease, but with peppermint sensitivity, positive prick test reactions to Mentha Piperita (Peppermint) Leaf,
Mentha Piperita (Peppermint) Leaf Extract, and Mentha Piperita were reported.72,73

Other Clinical Reports

Mentha Piperita (Peppermint) Oil

Positive reactions were observed in 7 of 450 dermatitic patients who were patch tested with 2% Mentha
Piperita (Peppermint) Oil in yellow soft paraffin.1 In another study, positive reactions to 2% Mentha Piperita
(Peppermint) Oil were observed in 6 of 86 dermatitic patients. A patch containing 1% Mentha Piperita
(Peppermint) Oil (vehicle unknown) was applied to the backs of 56 patients with chronic urticaria. No reactions
were noted after a 1-h or 48-h exposure.

No reactions were observed in 25 spice factory workers who were patch tested with 2% Mentha Piperita
(Peppermint) Oil in petrolatum. It has been reported that the patch testing of individual components of Mentha
Piperita (Peppermint) Oil using 3 patients with allergic contact dermatitis established that the allergens were
menthol and trace components such as piperitone or pulegone.1

Dermal
A triple-blind clinical trial involved 96 randomly selected subjects (47 cases and 49 controls; all pregnant
women) with a diagnosis of pruritus gravidarum.74 The case and control subjects were instructed to apply 60 ml of
peppermint oil (0.5% in sesame oil) and identical placebos, respectively, twice per day for 2 weeks. Applications
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were made to the area of the itch. The authors noted that Mentha Piperita (Peppermint) Oil did not cause any special
side effects in any of the subjects tested. They also stated that, based on these results, peppermint oil can be
consumed for symptomatic treatment of skin itching on pregnant women.

Oral
Each of 6 pediatric patients with irritable bowel syndrome received a single oral dose of Mentha Piperita
(Peppermint) Oil (187 mg).75 Each capsule contained 83 mg of menthol as a constituent of Mentha Piperita
(Peppermint) Oil. Each patient drank 125 ml of water after ingestion of the capsule. No adverse events were
reported. The delayed appearance of menthol in the plasma was reported; a substantial lag time (range 1 h to 4 h)
was observed in all subjects. Thus, an apparent prolonged absorption time was demonstrated. The authors noted that
reasons for the delayed time of peak (Tmax) likely related to formulation-specific factors (i.e., delayed release) and,
potentially, enterohepatic recirculation.

Exposure Assessment

Dermal
Mentha Piperita (Peppermint) Oil

The FDA calculated an estimated human exposure from cosmetic use based on the concentration of use
information supplied by industry.1 Using a body splash product containing 0.2% Mentha Piperita (Peppermint Oil)
and assuming 100% absorption over a body surface of 17,000 cm2 and a daily application of 1 mg/cm2 (~17 ml of
the product), the FDA estimated an exposure of 34 mg/day. For a 60-kg person, this amounted to an estimated daily
dose of 0.6 mg/kg/day.

Oral
Mentha Piperita (Peppermint) Oil

In the European Union, the highest recommended daily dose of Mentha Piperita (Peppermint) Oil is 1.2 ml,
i.e., 1080 mg Mentha Piperita (Peppermint) Oil (contains a maximum of 140 mg pulegone + menthofuran).16 For a
60 kg person, this would correspond to a daily intake of 2.3 mg/kg body weight. This recommended daily dose of
Mentha Piperita (Peppermint) Oil in medicinal products results in pulegone/menthofuran that exceeds the tolerated
daily intake (TDI) (0.1 mg/kg) that was established for food by the Committee of Experts on Flavoring Substances
(CEFS).

Risk Assessment

Dermal
Mentha Piperita (Peppermint) Oil

A maximum dermal use level of 5.4% has been recommended for Mentha Piperita (Peppermint) Oil.76 This
dermal restriction is based on 8% menthofuran (pulegone metabolite) and 3% pulegone content, with limits of 0.5%
for menthofuran and of 1.2% for pulegone. The authors also recommended that Mentha Piperita (Peppermint) Oil,
due to menthol content, should be avoided altogether in cases of cardiac fibrillation and in individuals with a
glucose-6-phosphate dehydrogenase deficiency. No further information relating to these recommendations is
provided.

Oral
Mentha Piperita (Peppermint) Oil and Pulegone (a component of Mentha Piperita (Peppermint) Oil)

The authors of a book entitled Essential Oil Safety have recommended a maximum daily oral dose of 152
mg Mentha Piperita (Peppermint) Oil.76 This oral restriction is based on 8% menthofuran and 3% pulegone content,
with limits of 0.2 mg/kg/day for menthofuran and 0.5 mg/kg/day for pulegone.
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Mentha piperita

A study was performed to characterize data on dietary botanical supplement (DBSs) associated with
adverse event reports submitted to the FDA Center for Food Safety and Applied Nutrition’s Adverse Event
Reporting System (CAERS).77 FDA obtained CAERS data from 1999 to 2003 involving adverse effects associated
with the 6 most frequently used DBSs, including peppermint. No adverse events were reported for single-ingredient
peppermint supplements during the study period.

SUMMARY

The safety of the following ingredients in cosmetics was previously reviewed by the Panel, and a final
report with a conclusion stating that these ingredients are safe as used in cosmetic formulations was published in
2001: Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Leaf Extract, Mentha Piperita (Peppermint)
Leaf, and Mentha Piperita (Peppermint) Leaf Water. The conclusion also stated that the concentration of pulegone, a
constituent of these botanical ingredients, should not exceed 1%. The current safety assessment is, in part, a re-
review of the 4 Mentha piperita (peppermint)-derived ingredients, and is inclusive of safety test data that have
become available since the final report was issued.

The current safety assessment is also an first review of the following 6 Mentha piperita (peppermint)-
derived ingredients: Mentha Piperita (Peppermint) Extract, Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract,
Mentha Piperita (Peppermint) Flower/Leaf/Stem Water, Mentha Piperita (Peppermint) Leaf Cell Extract, Mentha
Piperita (Peppermint) Leaf Juice, and Mentha Piperita (Peppermint) Meristem Cell Culture.

According to 2017 VCRP data, the greatest use frequency is being reported for Mentha Piperita
(Peppermint) Oil, which is being used in 827 cosmetic products, mostly leave-on products. The results of a
concentration of use survey provided in 2016 indicate that Mentha Piperita (Peppermint) Leaf Water is being used at
a concentration up to 40% in leave-on products, which is the greatest use concentration that is being reported for
Mentha piperita (peppermint)-derived ingredients reviewed in this safety assessment.

In the U.S., Mentha Piperita (Peppermint) Oil is GRAS for use in food for human consumption. It is also an
inactive ingredient in drug products that have been approved by the FDA, and is on the EPA list of active
ingredients eligible for minimum risk pesticide products.

Mentha Piperita (Peppermint) Leaf Extract (10% aqueous ethanol extract) caused a statistically significant
increase in the penetration of caffeine, but not salicylic acid, through porcine skin. Mentha Piperita (Peppermint)
Oil inhibited the penetration of benzoic acid through human skin.

Following oral administration, Mentha Piperita (Peppermint) Oil is relatively rapidly absorbed and
eliminated mainly via the bile. The major biliary metabolite is menthol glucuronide. Additional metabolites are
mono- or di-hydroxylated menthol derivatives.

Mentha Piperita (Peppermint) Leaf Extract (aqueous extract) was not nephrotoxic to rats when
administered (20 g/l) in drinking water daily for 30 days.

No adverse events were reported for single-ingredient peppermint supplements in a study that was
performed to characterize data on dietary botanical supplement (DBSs) associated with adverse event reports
submitted to the FDA Center for Food Safety and Applied Nutrition’s Adverse Event Reporting System (CAERS).

Mentha Piperita (Peppermint) Oil was clastogenic in a chromosome aberration test involving peripheral
blood lymphocytes. In a genotoxicity assay involving human lymphocytes, Mentha Piperita (Peppermint) Oil
induced sister chromatid exchanges in a dose-dependent manner. A trade name mixture containing 2.5% Mentha
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Piperita (Peppermint) Extract (diluted to a concentration of 10% (effective concentration of extract = 0.25%)) was
not cytotoxic or genotoxic in the Ames test, with or without metabolic activation.

Oral pretreatment with Mentha Piperita (Peppermint) Leaf Extract (aqueous extract) before exposure to
gamma radiation was found to be effective in protecting against chromosomal damage in the bone marrow of Swiss
albino mice. In another study, the oral administration of Mentha Piperita (Peppermint) Leaf extract had an
antigenotoxic (i.e., reduced the frequency of chromosomal aberrations and micronuclei in bone marrow cells) in
Swiss albino mice intraperitoneally injected with benzo(a)pyrene.

Oral dosing with Mentha Piperita (Peppermint) Leaf Extract caused a significant reduction in the number of
lung adenomas from an incidence of 67.92% in Swiss albino mice intraperitoneally injected with benzo[a]pyrene to
26.31%. Oral dosing with Mentha Piperita (Peppermint) Leaf Extract also caused inhibition of skin papilloma
formation induced by DMBA and the application of croton oil, in terms of a significant decrease in the cumulative
number of papillomas, tumor burden, and tumor incidence.

In the human cancer cell lines tested with Mentha Piperita (Peppermint) Leaf Extract (various extractants
used), the number of apoptotic cells was incremental with an increase in the dose of Mentha piperita extracts.
Mentha Piperita (Peppermint) Leaf Extract (only from chloroform and ethyl acetate extractants) had significant
dose- and time-dependent anticarcinogenic activity, leading to G1 cell cycle arrest and mitochondrial-mediated
apoptosis, perturbation of oxidative balance, upregulation of Bax gene, and elevated expression of p53 and p21 in
the treated cells. In a study that was performed to evaluate the antitumor activity of Mentha Piperita (Peppermint)
Leaf Extract (methanol extract) in an anti-proliferation assay involving SW-480 human colon adenocarcinoma cells,
statistically significant growth inhibition was observed.

Results were positive for Mentha Piperita (Peppermint) Oil (from plants harvested during seasons of the
year) in cytotoxicity assays involving human cancer cell lines: The following IC50 values (µg/ml) were reported:
MCF-7 cell line (75.2 ± 2.9 [summer]; 80.8 ± 3.2 [winter]) and LNCaP cell line (90.4 ± 3.7 [summer]; 95.7 ± 4.5
[winter]). In another study, Mentha Piperita (Peppermint) Oil was cytotoxic to the following human cancer cell
lines: human lung carcinoma SPC-A1 cells (IC50 = 10.89 µg/ml), human leukemia K562 cells (IC50 = 16.16 µg/ml)
and human gastric cancer SGC-7901 cells (IC50 = 38.76 µg/ml). The essential oil was inactive against human
hepatocellular carcinoma BEL-7402 cells.

Mentha Piperita (Peppermint) Leaf Extract (1000 µg/ml, methanol extract) did not induce hepatotoxicity in
in vitro assays involving human (HepG2/C3A) and rat (MH1C1) hepatoma cells.

The 50% ethanol extract of peppermint leaves and stems significantly inhibited compound 48/80-induced
histamine release from rat peritoneal mast cells in vitro.

In a study involving C57BL/6 mice, it was concluded that 3% Mentha Piperita (Peppermint) Oil (diluted in
jojoba oil) facilitated hair growth by promoting the conservation of vascularization of hair dermal papilla.

A trade name mixture containing 7.5% Mentha Piperita (Peppermint) Extract was non-irritating to the skin
of 3 rabbits. No macroscopic cutaneous reactions attributable to allergy were observed in a maximization test in
which 10 albino guinea pigs were patch tested with a trade name material containing 7.5% Mentha Piperita
(Peppermint) Extract during induction and challenged with the undiluted material and the material at a concentration
of 50% (effective concentration of extract = 3.75%)

In a 48-h occlusive patch test, a lipstick product containing 0.2961% Mentha Piperita (Peppermint) Leaf
Extract did not cause skin irritation in any of the 50 subjects tested. In an in vitro skin irritation study on Mentha
Piperita (Peppermint) Leaf Extract (water/ethanol extract) involving reconstructed human epidermis, results were
negative (MTT > 50%).

There was no evidence of dermal irritation or allergic contact sensitization in an HRIPT in which 52 male
and female subjects were patch tested with a 10% dilution of a trade name mixture containing 2.5% Mentha Piperita
(Peppermint) Extract (effective concentration of extract = 0.25%). A cosmetic product containing 0.00554%
Mentha Piperita (Peppermint) Extract did not cause dermal irritation or allergic contact dermatitis in 51 male and
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female subjects patch tested with a cosmetic product (an off-white cream) containing 0.00554% Mentha Piperita
(Peppermint) Extract. In the maximization test, a cosmetic product (off-white cream) containing 0.00554% Mentha
Piperita (Peppermint) did not induce contact sensitization in the 26 male and female subjects tested.

A face cream containing 20% Mentha Piperita (Peppermint) Leaf Water did not induce cumulative skin
irritation or sensitization in an HRIPT involving 107 subjects. Negative results were also reported for a lipstick
product containing 0.2961% Mentha Piperita (Peppermint) Leaf Extract in a 48-h occlusive patch test (skin irritation
test) involving 50 subjects. Slight erythema was observed in 1 of 50 subjects who applied a cleansing gel containing
50% Mentha Piperita (Peppermint) Leaf Water twice per day for 4 weeks; there were no signs of ocular or palpebral
irritation in any of the subjects. In a 48-h, single-application patch test, the skin irritation potential of a cleansing gel
containing 50% Mentha Piperita (Peppermint) Leaf Water (10% aqueous dilution [effective concentration = 5%])
was evaluated using 52 subjects. A score of 1 (mild erythema) was reported for 12 subjects on day 2 and for 18
subjects on day 3. A score of 2 (moderate erythema) was reported for 2 subjects on day 2 and for 3 subjects on day
3. On day 4, 46 subjects had a score of 0 and 6 had a score of 1. The authors concluded that the skin compatibility of
the diluted product was considered good.

The skin irritation and sensitization potential of 20% Mentha Piperita (Peppermint) Oil was evaluated in an
HRIPT involving 104 subjects and results were negative. Similarly, there was no evidence of sensitization to 20%
Mentha Piperita (Peppermint) Oil in an HRIPT involving 101 subjects. HRIPT results for undiluted Mentha Piperita
(Peppermint) Leaf Extract (water/ethanol extract) in 52 subjects were also negative.

A trade name mixture containing 2.5% Mentha Piperita (Peppermint) Extract was classified as non-
irritating in an in vitro toxicity testing system, consisting of normal, human-derived epidermal keratinocytes, for
evaluating ocular irritation potential. Slight ocular irritation was observed in a study in which 3 rabbits were tested
with a trade name mixture containing 7.5% Mentha Piperita (Peppermint) Extract.

In a multicenter study, neither irritant nor allergic reactions were observed in 73 patients patch tested with
Mentha Piperita (Peppermint) Oil according to International Contact Dermatitis Research Group (ICDRG) patch test
procedures. Another multicenter study involving 13,398 patients was performed by the US/Canadian North
American Contact Dermatitis Group (NACDG), whereby 71 patients were tested with Piperita (Peppermint) Oil (2%
in petrolatum). The prevalence rate for this ingredient was 0.53 %.

Mostly positive patch test reactions to Mentha Piperita (Peppermint) Oil (2%in petrolatum) and Mentha
Piperita were reported in case reports on patients with diseased skin. In patients without skin disease, but with
peppermint sensitivity, positive prick test reactions to Mentha Piperita (Peppermint) Leaf, Mentha Piperita
(Peppermint) Leaf Extract, and Mentha Piperita were reported. In other clinical reports, oral dosing with Mentha
Piperita (Peppermint) Oil (0.5% in sesame oil, 60 ml) did not cause any side effects in 47 pregnant female patients.
A single oral dose of Mentha Piperita (Peppermint) Oil did not cause any adverse events in 6 pediatric patients.

DISCUSSION

The safety of the following cosmetic ingredients was reviewed previously by the Panel, and a final report
with a conclusion stating that these ingredients are safe as used in cosmetic formulations was published in 2001:
Mentha Piperita (Peppermint) Oil, Mentha Piperita (Peppermint) Leaf Extract, Mentha Piperita (Peppermint) Leaf,
and Mentha Piperita (Peppermint) Leaf Water. The conclusion also states that the concentration of pulegone, a
constituent of these botanical ingredients, should not exceed 1%. The current safety assessment is a re-review of the
safety of these 4 ingredients, and is also an initial safety evaluation of the following 6 related ingredients that were
not listed as cosmetic ingredients prior to development of the published safety assessment: Mentha Piperita
(Peppermint) Extract, Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract, Mentha Piperita (Peppermint)
Flower/Leaf/Stem Water, Mentha Piperita (Peppermint) Leaf Cell Extract, Mentha Piperita (Peppermint) Leaf Juice,
and Mentha Piperita (Peppermint) Meristem Cell Culture.

In the 2001 published final safety assessment on Mentha piperita (peppermint)-derived ingredients, the
Panel expressed concern that rat oral-dosing studies on Mentha Piperita (Peppermint) Oil and pulegone reported
cyst-like spaces in the cerebellum that were attributed to pulegone. Therefore, the Panel established a 1%
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concentration limit on pulegone in Mentha piperita (peppermint)-derived ingredients due to toxicity concerns. In
these studies, brain sections were fixed by immersion using 4% neutral buffered formaldehyde. Because immersion
fixation of nervous tissue might cause artifacts observed as vacuolar retraction spaces around neurons, a study
involving rats was performed, using both immersion and perfusion tissue fixation methods, to determine whether the
cerebellar lesions observed in earlier studies were caused by dosing with pulegone. Study results for rats dosed
with pulegone did not reveal the occurrence of test substance-related, cyst-like spaces in the white matter of the
cerebellum using either perfusion or immersion tissue fixation techniques. A possible explanation for the observed
dose-dependent cyst-like spaces seen in previous studies could have been due to an interaction between impurities in
the test substance and the fixation agent used therein. Given this possibility, the Panel agreed that the brain lesions
may have been an artifact of the fixation method, and that the 1% limitation on pulegone is no longer warranted.

The Panel also considered the positive effects that were observed in female rats, and in male and female
mice, dosed with pulegone in the 2011 National Toxicology Program (NTP) oral carcinogenicity study. However,
the Panel did not express concern over these findings relative to pulegone as a component of Mentha Piperita
(Peppermint) Oil in cosmetic products, based on the understanding that the cytotoxic dose-response relationship
(renal and liver toxicity) that was associated with cancer development would not be relevant to pulegone exposure
from a cosmetic product containing Mentha piperita (Peppermint) Oil at current use concentrations.

Because final product formulations may contain multiple botanicals, each possibly containing the same
constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching levels that
may be hazardous to consumers. For Mentha piperita (peppermint)-derived ingredients, the Panel is concerned
about the presence of terpenes (e.g., limonene) and terpenoids (e.g. menthol) in cosmetics, which could result in
sensitization.

The Panel expressed concern about pesticide residues, heavy metals, and other plant species that may be
present in botanical ingredients. They stressed that the cosmetics industry should continue to use current good
manufacturing practices (cGMPs) to limit impurities.

Additionally, the Panel recognized that Mentha Piperita (Peppermint) Leaf Extract can enhance the
penetration of other ingredients through the skin. The Panel cautioned that care should be taken in formulating
cosmetic products that may contain Mentha piperita (peppermint)-derived ingredients in combination with any
ingredients whose safety was based on their lack of dermal absorption data, or when dermal absorption was a
concern.

The issue of incidental inhalation exposure was discussed by the Panel, as Mentha piperita (peppermint)-
derived ingredients are being used in products that could possibly be inhaled. For example, Mentha Piperita
(Peppermint) Leaf Extract is used in face and neck sprays at maximum use concentrations up to 1.3%. Another
example relating to inhalation exposure from products that are sprayed is the use of Mentha Piperita (Peppermint)
Oil in both pump hair sprays (maximum use concentrations up to 0.02%) and aerosol hair sprays (maximum use
concentrations up to 0.017%) which may result in incidental inhalation exposure. The use of Mentha Piperita
(Peppermint) Leaf Extract at maximum use concentrations up to 0.0018% in face powders may also result in
incidental inhalation exposure. Additionally, the Panel noted that droplets/particles from spray cosmetic products
would not be respirable to any appreciable amount. Furthermore, droplets/particles deposited in the nasopharyngeal
or bronchial regions of the respiratory tract present no toxicological concerns based on the chemical and biological
properties of these ingredients. Coupled with the small actual exposure in the breathing zone and the concentrations
at which the ingredients are used, the available information indicates that incidental inhalation would not be a
significant route of exposure that might lead to local respiratory or systemic effects. A detailed discussion and
summary of the Panel’s approach to evaluating incidental inhalation exposures to ingredients in cosmetic products is
available at http://www.cir-safety.org/cir-findings.

Finally, the Panel agreed that the available composition data on seven of the ingredients are sufficient, but the
data relating to composition of Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract, Mentha Piperita
(Peppermint) Flower/Leaf/Stem/Water, and Mentha Piperita (Peppermint) Meristem Cell Culture are insufficient.
Also, after considering the available skin irritation and sensitization data, the Panel determined that skin
sensitization data on these three ingredients also are insufficient. Thus, the Panel determined that the following
additional data are needed in order to evaluate the safety :
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• Composition data on Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract, Mentha Piperita (Peppermint)
Flower/Leaf/Stem/Water and Mentha Piperita (Peppermint) Meristem Cell Culture.
○ Depending on the composition data that are received, other toxicological endpoints may be needed.

• Skin irritation and sensitization data on Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract, Mentha
Piperita (Peppermint) Flower/Leaf/Stem/Water and Mentha Piperita (Peppermint) Meristem Cell Culture.

The Panel also agreed that the available data are sufficient for determining that Mentha Piperita (Peppermint) Oil,
Leaf, and leaf-derived ingredients are safe in cosmetics in the present practices of use and concentration, when
formulated to be non-sensitizing. The non-sensitizing caveat relates to the avoidance of a cumulative effect of
multiple botanicals, in a single formulation, that share a constituent in common.

CONCLUSION

The CIR Expert Panel concluded that the following 7 Mentha piperita (peppermint)-derived ingredients are
safe in cosmetics in the present practices of use and concentration described in the safety assessment, when
formulated to be non-sensitizing:
• Mentha Piperita (Peppermint) Oil • Mentha Piperita (Peppermint) Leaf Extract
• Mentha Piperita (Peppermint) Extract • Mentha Piperita (Peppermint) Leaf Juice*
• Mentha Piperita (Peppermint) Leaf • Mentha Piperita (Peppermint) Leaf Water
• Mentha Piperita (Peppermint) Leaf Cell Extract*

*Not reported to be in use. Were ingredients in this group not in current use to be used in the future, the expectation is that
they would be used in product categories and at concentrations comparable to others in this group.

The CIR Expert Panel also concluded that the available data are insufficient to make a determination that the
following 3 Mentha piperita (peppermint)-derived ingredients are safe under the intended conditions of use in cosmetic
formulations:

• Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract**

• Mentha Piperita (Peppermint) Flower/Leaf/Stem Water**
• Mentha Piperita (Peppermint) Meristem Cell Culture**

**Not reported to be in use.

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TABLES

Table 1. Toxicity Data on Components of Mentha Piperita (Peppermint)–Derived Ingredients

Study Type/Protocol Results

Short-Term Oral Toxicity

Menthone: 28-day study on groups of 20 rats. Doses up to 800 Brain lesions (cyst-like spaces in the cerebellum).1
mg/kg/day.
Menthol: 28-day study on groups of 20 rats. Doses up to 800 No brain lesions.1
mg/kg/day
Pulegone: 28-day study on groups of 20 rats. Doses up to 160 Brain lesions (cyst-like spaces in the cerebellum).1
mg/kg/day
Pulegone: 28-day study on groups of 28 female rats (CrL: (WI)BR, Alopecia. Statistically significant, test substance-related decrease in
SPF strain). Doses (in soybean oil) of 160 mg/kg/day. Untreated body weight and food consumption (p < 0.001) observed throughout t
control. study. Statistically significant reduction in plasma creatinine.
Statistically significant increase in plasma alkaline phosphatase and
non-statistically significant increase in alanine aminotransferase.
Increased plasma concentrations of alkaline phosphatase and alanine
aminotransferase taken together with increased absolute liver weight
was non- statistically significant, but increased relative liver weight (p
< 0.05) was indicative of adverse effect on the liver. However, no
significant histopathology of liver observed. Number and severity of
cyst-like spaces in cerebellum considered comparable to observations
normally present in historical control Wistar rats at laboratory where
study was performed. Authors noted that dose-dependent cyst- like
spaces in cerebellum observed in previous studies could have been due
to interaction between impurities in test substance and fixation agent
used. 4

Developmental and Reproductive Toxicity

Menthol: Groups of 15 to 23 pregnant animals dosed with Brazilian No teratogenic effects. 1

menthol via oral intubation. Mice received doses up to 185 mg/kg/day
on gestation days (GD) 6 to 15. Rats received doses up to 218 mg/kg
on GDs 6 to 15. Hamsters received doses up to 405 mg/kg/day on
GDs 6 to 10. Artificially inseminated rabbits received doses up to 425
mg/kg/day on GDs 6 to 18. Caesarean sections were performed on all
dams.

Genotoxicity

Menthol: Salmonella typhimurium strains: TA1535, TA100, TA1537, Toxicity at highest dosed; less toxicity with metabolic activation.
and TA98. Doses of 6.4, 32, 160, and 800 µg/plate in Ames test with Same number of revertants reported for test and control cultures
or without metabolic activation. exposed to lower doses.1
Pulegone: Salmonella typhimurium strains: TA1535, TA100, TA1537, Toxicity at highest dose; less toxicity with metabolic activation. Same
and TA98. Doses of 6.4, 32, 160, and 800 µg/plate in Ames test with number of revertants reported for test and control culture exposed to
or without metabolic activation. lower doses.1
Menthone: Salmonella typhimurium strains: TA1535, TA100, Toxicity at highest dose. Statistically significant number of revertants
TA1537, TA98, and TA97. Doses of 6.4, 32, 160, and 800 µg/plate in in strain TA1537 at doses of 6.4 and 32 µg/plate without metabolic
Ames test with or without metabolic activation. activation. Further testing with a more sensitive strain (TA98) yielded
statistically significant increases in the number of revertants at all
doses tested without metabolic activation; results were dose-related.1

Menthol (Brazilian): Cytogenetic assay (rats). Doses of 1.45, 14.5, Non-genotoxic.1

and 145 mg/kg, and, in some instances, doses of 500, 1150, and 3000
mg/kg, or 5000 mg/kg.
Menthol (Brazilian): Dominant lethal assay (rats). Doses of 1.45, Non-genotoxic.1
14.5, and 145 mg/kg, and, in some instances, doses of 500, 1150, and
3000 mg/kg, or 5000 mg/kg.
Menthol (Brazilian): Host-mediated assay (mice). Doses of 1.45, Weakly positive but significant response at high dose in Salmonella
14.5, and 145 mg/kg, and, in some instances, doses of 500, 1150, and typhimurium TA1530, and elevated recombinant frequencies noted in
3000 mg/kg, or 5000 mg/kg. Salmonella typhimurium strain TA1530 Saccharomyces strain D3.1
and Saccharomyces strain D3 tested in vitro.

Carcinogenicity

Menthol: National Cancer Institute (NCI) 2-year bioassay. dl- Negative trend in fibroadenomas of the mammary gland observed in
menthol administered orally to Fischer 344 rats (3750 ppm or 7500 female rats (20 of 50 control; 10 of 49 low-dose; 7 of 49 high-dose).
ppm) or to B6C3F1 mice (2000 ppm or 4000 ppm). No evidence of carcinogenicity in rats or mice or either sex.1
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Table 1. Toxicity Data on Components of Mentha Piperita (Peppermint)–Derived Ingredients

Study Type/Protocol Results

Carcinogenicity

Pulegone: National Toxicology Program (NTP) 2-year bioassay. Effects in the kidneys (hyaline glomerulopathy and nephropathy), liver
Pulegone (in corn oil) administered to groups of 50 male and 50 (oval cell hyperplasia, bile duct hyperplasia, hypertrophy, hepatocyte
female F344/N rats and groups of 50 male and 50 female B6C3F1 necrosis, and portal fibrosis), nose (olfactory epithelium degeneration,
mice by gavage (5 days/week) for 105 weeks. Male rats received inflammation, and metaplasia), and forestomach (inflammation,
doses of 18.75, 37.5, or 75 mg/kg; female rats and male and female hyperplasia, and ulcer) reported. Increased incidences of liver
mice received doses of 37.5, 75, or 150 mg/kg. neoplasms in male and female B6C3F1 mice in the study led to the
conclusion that there was clear evidence of carcinogenic activity in
mice. For female F344/N rats, it was concluded that there was some
evidence of carcinogenicity based on an increased incidence of urinary
bladder neoplasms. Five of 47 rats in 150 mg/kg female stop-exposure
group (gavage with pulegone stopped at week 60 because of severely
reduced body weights) diagnosed with papilloma and carcinoma,
combined. Male rats did not show increased incidences of bladder
tumors or neoplasms of other organs.3 The International Agency for
Research on Cancer (IARC) concluded that there is sufficient evidence
in experimental animals for the carcinogenicity of pulegone, and that
there is inadequate evidence in humans for the carcinogenicity of
pulegone.78 IARC’s conclusions were based on positive oral
carcinogenicity data in male and female mice and in female rats.

Pulegone: Female rats dosed orally (gavage) with 75 or 150 mg/kg for Results supported hypothesis that cytotoxicity followed by
4 and 6 weeks. regenerative cell proliferation is mode-of-action for pulegone-induced
urothelial tumors in female rats.79
Anticarcinogenicity

(-)-Menthol: Rats dosed orally with 1%or 0.5% (-)-menthol for 20 Significant inhibition (p < 0.001) of DMBA-induced rat mammary
weeks. Dosing initiated 2 weeks prior to dimethylbenzanthracene gland carcinogenesis following 20 weeks of oral dosing with 1% (-)-
(DMBA) tumor induction. menthol. Chemopreventive effect noted when rats dosed with 0.5%
menthol for 2 weeks prior to and 1 week after DMBA induction.1

Skin Irritation

Methanol: Two Tiger Balm formulations containing 8% and 10% Dermal irritation observed in all treated animals, with the following
menthol applied for 23 h, under occlusive patches, to abraded and severity scale: 8% menthol balm < control wax < 10% menthol balm.
intact sites on New Zealand white rabbits. Total number of patches 8% menthol balm almost innocuous in male rabbits. Irritation
applied was 21. A third group was treated with control wax (mixture observed was not progressive and tolerance developed within 10 days.
of hard and soft waxes). No severe damage noted at microscopic examination of skin (increased
hyperkeratosis noted at treated sites), and no evidence of systemic
toxicity.1

Skin Sensitization

Menthol Because menthol is a predominant component of Mentha Piperita

(Peppermint) Oil and Mentha Piperita (Peppermint) Leaf Extract, it
should be noted that a review article on the sensitization potential of
menthol is available. After performing an extensive review of the
reported cases of sensitization to menthol, it was concluded that the
literature does not adequately document the clinical relevance of the
patch test reactions nor the impact of irritant reactions. 80

Other Clinical Reports

Menthol: 877 patients with primary contact, atopic, nummular, and Reactions observed in 1% of the panelists within 96 h.1
stasis dermatitis and eczema were tested with 5% menthol in yellow
paraffin
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Table 2. Definitions and Functions of the Ingredients in this Safety Assessment.2

Ingredient CAS No. Definition & Idealized Structures Function
Mentha Piperita (Peppermint) Oil Mentha Piperita (Peppermint) Oil is a volatile oil obtained from the Fragrance Ingredients;
8006-90-4 whole plant Mentha piperita. The accepted scientific name for Mentha Skin-Conditioning Agents
84082-70-2 piperita is Mentha x piperita. - Miscellaneous

Mentha Piperita (Peppermint) Leaf Extract Mentha Piperita (Peppermint) Leaf Extract is the extract of the leaves of Fragrance Ingredients;
84082-70-2 the peppermint, Mentha piperita. The accepted scientific name for Skin-Conditioning Agents
Mentha piperita is Mentha x piperita. - Miscellaneous; Skin-
Conditioning Agents -
Occlusive

Mentha Piperita (Peppermint) Leaf Mentha Piperita (Peppermint) Leaf is the dried leaves of Mentha Fragrance Ingredients
piperita. The accepted scientific name for Mentha piperita is Mentha x
piperita.

Mentha Piperita (Peppermint) Leaf Water Mentha Piperita (Peppermint) Leaf Water is an aqueous solution of the Flavoring Agents;
84082-70-2 steam distillate obtained from the leaves of Mentha piperita. The Fragrance Ingredients;
accepted scientific name for Mentha piperita is Mentha x piperita. Skin-Conditioning Agents
- Miscellaneous

Mentha Piperita (Peppermint) Extract Mentha Piperita (Peppermint) Extract is the extract of the whole plant, Skin-Conditioning Agents
84082-70-2 Mentha piperita. The accepted scientific name for Mentha piperita is - Miscellaneous
Mentha x piperita.

Mentha Piperita (Peppermint) Mentha Piperita (Peppermint) Flower/Leaf/Stem Extract is the extract of Flavoring Agents;
Flower/Leaf/Stem Extract the flowers, leaves and stems of Mentha piperita. The accepted Fragrance Ingredients;
84082-70-2 scientific name for Mentha piperita is Mentha x piperita. Skin-Conditioning Agents
- Miscellaneous

Mentha Piperita (Peppermint) Mentha Piperita (Peppermint) Flower/Leaf/Stem Water is the aqueous Fragrance Ingredients
Flower/Leaf/Stem Water solution of the steam distillates obtained from the flowers, leaves and
84082-70-2 stems of Mentha piperita. The accepted scientific name for Mentha
piperita is Mentha x piperita.

Mentha Piperita (Peppermint) Leaf Cell Mentha Piperita (Peppermint) Leaf Cell Extract is the extract of a Antioxidants; Skin
Extract culture of the leaf cells of Mentha piperita. The accepted scientific Protectants
name for Mentha piperita is Mentha x piperita.

Mentha Piperita (Peppermint) Leaf Juice Mentha Piperita (Peppermint) Leaf Juice is the juice expressed from the Skin-Conditioning Agents
84082-70-2 leaves of Mentha piperita. The accepted scientific name for Mentha - Miscellaneous
piperita is Mentha x piperita.

Mentha Piperita (Peppermint) Meristem Cell Mentha Piperita (Peppermint) Meristem Cell Culture is a suspension of Skin-Conditioning Agents
Culture the cultured meristem cells of Mentha piperita. The accepted scientific - Miscellaneous
name for Mentha piperita is Mentha x piperita.
 Distributed for comment only -- do not cite or quote

Table 3. Physical and Chemical Properties of Mentha Piperita (Peppermint)-Derived Ingredients

Property Value Reference
Mentha Piperita (Peppermint) Oil
1
Form Colorless or pale yellow liquid
Angular rotation (˚C) Between -18 and 32 1

Refractive index (at 20˚C) Between 1.459 and 1.465 1

1
Specific gravity Between 0.896 and 0.908
1
Assay for total esters Not less than 5% of esters, calculated as menthyl acetate
1
Assay for total menthol Not less than 50% of menthol
1
Dimethyl sulfide Passes test (rectified); fails test (natural)
1
Heavy metals (as Pb) Passes test (limit of 0.004%)
1
Solubility Soluble in alcohol: Passes test (1 volume dissolves in 3 volumes of 70% alcohol);
7
Soluble in most vegetable oils; insoluble in propylene glycol

Mentha Piperita (Peppermint) Leaf Extract

5
Form Yellow to light brown, clear oil
Refractive index (at 20˚C) 1459-1469 5

Density (at 20˚C) 0.890-0.910 5

Mentha Piperita (Peppermint) Extract (2.5% in tradename mixture)

6
Refractive index 1.4498
6
Specific gravity 1.031
 Distributed for comment only -- do not cite or quote

Table 4. Chromatographic Profiles for Mentha Piperita (Peppermint) Oil15 and Mentha Piperita (Peppermint) Leaf Extract*.5,19
Origins Other Than U.S. U.S. Type
Components Min. Max. Min. Max.
(%) (%) (%) (%)
Mentha Piperita
(Peppermint) Oil
3-Octanol 0.1 0.5 0.1 0.4
1,8-Cineol 3.0 8.0 4.0 6.0
Limonenea 1.0 3.0 1.0 2.5
trans-Sabinene Hydrate 0.5 2.0 0.5 2.3
Menthone 13.0 28.0 15.0 25.0
Isomenthone 2.0 8.0 2.0 4.5
Menthofuran 1.0 8.0 1.5 6.0
Neomenthol 2.0 6.0 2.5 4.5
Menthol 32.0 49.0 36.0 46.0
Pulegone 0.5 3.0 0.5 2.5
Menthyl Acetateb 2.0 8.0 3.0 6.5
β-Caryophyllene 1.0 3.5 1.0 2.5
Origin Not Stated
Mentha Piperita
(Peppermint) Leaf Extract
Essential Oilc 80 90
Essential Oild > 65
Water Not specified

Volatile Compoundse
Limonene <2
1,8-Cineol not specified
L-Menthone 20 40
Menthofuran <2
Isomenthone Not specified
Isomenthol Not specified
Neomenthol Not specified
L-Menthol 25 40
Pulegone <3
Menthyl Acetate 5 15
beta Caryophyllene Not specified
Germacrene Not specified

Allergen Compoundsf
Eugenol < 0.05
Linalool < 0.3
d-Limonene <1
*Mentha Piperita (Peppermint) Leaf Extract (CO2 extract)
a
The limonene is regarded to be predominantly L-limonene based on physical tests. It is believed that there might be a small amount of D-limonene present,
but the exact quantity is unknown
b
The menthyl acetate is regarded to be predominantly L-menthyl acetate based on the physical tests. It is believed that there might be a small amount of D-
menthyl acetate present, but the exact quantity is unknown.
C
Gravimetric distillation detection method used
d
Volumetric distillation detection method used
e
Gas chromatography-mass spectrometry detection method used
f
Allergen compounds present are subject to declaration if the concentration exceeds 0.001% in leave-on and 0.01% in rinse-off products. However, values <
0.01% are not mentioned.
 Distributed for comment only -- do not cite or quote

Table 5. Frequency and Concentration of Use of Mentha piperita (peppermint)-derived Ingredients According to Duration and Exposure. 1,24,25
# of Uses Max Conc of Use (%) # of Uses Max Conc of Use (%)
Mentha Piperita (Peppermint) Oil Mentha Piperita (Peppermint) Leaf Extract
2017 1998 2017 1997 2017 1998 2017 1997
Totals* 827 102 0.0001-5 0.1-3 198 35 0.000075-0.5 NR
Duration of Use
Leave-On 433 52 0.0006-5 0.2-2 118 10 0.000075-0.5 NR
Rinse-Off 360 44 0.0001-1.9 0.1-3 79 24 0.0001-0.2 NR
Diluted for (Bath) Use 34 6 0.018-3.9 NR 1 1 NR NR
Exposure Type
Eye Area 4 NR 0.00094 NR 4 NR 0.0018 NR
Incidental Ingestion 214 24 0.05-2.9 0.2-1.2 7 NR 0.00075-0.5 NR
Incidental Inhalation-Spray 19;120a NR;23a 0.017-1;0.002- NR 5;34a NR;2a 0.00041- NR
1.1a 0.0046;0.00057-
0.026a
Incidental Inhalation-Powder 1 NR 0.01-1 NR 3 NR 0.0018 NR
Dermal Contact 458 75 0.0006-5 0.1-2 139 22 0.000075-0.2 NR
Deodorant (underarm) 2 NR NR NR NR NR 0.0018 NR
Hair - Non-Coloring 148 1 0.0001-0.96 3 50 13 0.00018-0.2 NR
Hair-Coloring NR NR 0.0024 NR NR NR 0.032 NR
Nail 7 2 0.00064-1.5 NR 2 NR NR NR
Mucous Membrane 317 30 0.0025-3.9 0.2-1.2 8 6 0.00075-0.5 NR
Baby Products 1 NR 0.2 NR NR NR NR NR
Mentha Piperita (Peppermint) Leaf Mentha Piperita (Peppermint) Leaf Water
2017 1998 2017 1997 2017 1998 2017 1997
Totals* NR NR 0.0002-1.6 NR 15 NR 0.00013-40 NR
Duration of Use
Leave-On NR NR 0.001 NR 9 NR 0.00013-40 NR
Rinse-Off NR NR 0.0002-1.6 NR 6 NR 0.00021-0.00067 NR
Diluted for (Bath) Use NR NR NR NR NR NR NR NR
Exposure Type
Eye Area NR NR NR NR 2 NR NR NR
Incidental Ingestion NR NR NR NR NR NR NR NR
Incidental Inhalation-Spray NR NR NR;0.001a NR NR;4a NR NR;0.00043-10a NR
Incidental Inhalation-Powder NR NR NR NR NR NR NR NR
Dermal Contact NR NR 0.001-1.6 NR 13 NR 0.00013-40 NR
Deodorant (underarm) NR NR NR NR 1 NR 15 NR
Hair - Non-Coloring NR NR 0.0002-0.023 NR NR NR NR NR
Hair-Coloring NR NR NR NR NR NR NR NR
Nail NR NR NR NR NR NR NR NR
Mucous Membrane NR NR 0.001-0.16 NR NR NR NR NR
Baby Products NR NR NR NR NR NR NR NR
Mentha Piperita (Peppermint) Extract
2017 2017
Totals* 71 0.00006-7.9
Duration of Use
Leave-On 43 0.00006-7.9 NR
Rinse-Off 27 0.0001-1 NR
Diluted for (Bath) Use 1 1 NR
Exposure Type
Eye Area NR NR NR
Incidental Ingestion 4 0.0099-3.4 NR
Incidental Inhalation-Spray 10; 21a 1.3; 0.005-1a NR
Incidental Inhalation-Powder NR NR NR
Dermal Contact 63 0.00006-7.9 NR
Deodorant (underarm) 1 1 NR
Hair - Non-Coloring 4 0.0004-1 NR
Hair-Coloring NR NR NR
Nail NR NR NR
Mucous Membrane 3 0.0099-3.4 NR
Baby Products NR NR NR
*Because each ingredient may be used in cosmetics with multiple exposure types, the sum of all exposure types may not equal the sum of total uses.
a
It is possible these products are sprays, but it is not specified whether the reported uses are sprays..
NR - no reported use
 Distributed for comment only -- do not cite or quote

Table 6. Dermal Irritation and Sensitization Data

Ingredient Subjects/Cell Type Protocol Results

Irritation Studies

In Vitro
Mentha Piperita Reconstructed human In vitro reconstructed human Negative (MTT > 50%).11
(Peppermint) Leaf Extract epidermis epidermis test method. In this
(water/ethanol extract) (10% test, the reduction of 3-(4,5-
and 100%) dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide
(MTT) in test material-treated
tissues is expressed as a
percentage relative to negative
control-treated cultures.

Animal
Trade name mixture 3 rabbits (strain not Mixture (0.5 ml) applied under No cutaneous reactions
containing 7.5% Mentha stated) semi-occlusive dressing to observed. Classified as non-
Piperita (Peppermint) intact skin for 4 h. Area of irritant.9
Extract application (cm2) not stated.

Human
Lipstick containing 50 subjects, described as In occlusive patch test, product Product and negative control did
0.2961% Mentha Piperita follows: normal (25), (~ 20 mg) applied in a square not cause skin irritation in any
(Peppermint) Leaf Extract with eczema (4 test chamber (8 mm x 8 mm) to of the subjects tested and
subjects), with allergy (4 the back of each subject for 48 product was classified as
subjects) and with h. Reactions scored 30 harmless. Positive control
sensitive skin (17 minutes after patch removal caused positive reactions in 12
subjects). and at 72 h post-application. subjects.53
Sodium dodecyl sulfate (1% in
water) and water served as the
positive and negative controls,
respectively.

50% Mentha Piperita 50 subjects Subjects applied product twice Slight erythema was observed in
(Peppermint) Leaf Water in per day for 4 weeks, and were one subject.54
a cleansing gel instructed to record any signs
felt or observed during product
use.

50% Mentha Piperita 52 subjects Diluted product applied, under A score of 1 (mild erythema)
(Peppermint) Leaf Water in occlusive patch, to the skin for was reported for 12 subjects on
a cleansing gel (10% 48 h. Reactions scored up to day 2 and for 18 subjects on day
aqueous dilution [effective 48 ± 4 h after patch removal 3. A score of 2 (moderate
concentration = 5%]) (day 4). erythema) was reported for 2
subjects on day 2 and for 3
subjects on day 3. On day 4, 46
subjects had a score of 0 and 6
had a score of 1. Skin
compatibility of diluted product
considered good.55
Sensitization Studies
Animal 10 albino guinea pigs Maximization test. 1st No macroscopic cutaneous
Trade name material induction: 2 intradermal reactions attributable to allergy
containing 7.5% Mentha injections of trade name associated with application of
Piperita (Peppermint) material, 2 injections of material. No cutaneous
Extract Freund’s complete adjuvant intolerance reactions in animals
(FCA), and 2 injections of of the negative control group
FCA and material mixture. 2nd (further details not provided).9
induction: topical application
of material 24 h after brushing
with 10% sodium lauryl sulfate
(SLS). After 19-day non-
treatment period, challenge
phase involved topical
applications of material
(undiluted and at concentration
of 50% (effective concentration
of extract = 3.75%)) under an
occlusive dressing for 24 h
 Distributed for comment only -- do not cite or quote

Table 6. Dermal Irritation and Sensitization Data

Ingredient Subjects/Cell Type Protocol Results

Human
Trade name mixture 52 male and female Human repeated insult patch No evidence of dermal irritation
containing 2.5% Mentha subjects. test (HRIPT). 1" x 1" semi- or allergic contact sensitization
Piperita (Peppermint) occlusive patch containing the in any of the subjects tested.
Extract. Prior to application, diluted mixture (0.2 ml) Also, no adverse events were
mixture prepared as a 10% applied for 24 h to upper back identified during the study.58
dilution using distilled water (between the scapulae) 3 times
(effective concentration of per week for total of 9
extract = 0.25%). applications. After a 2-week
non-treatment period, diluted
mixture applied to new test site
adjacent to original site.
Reactions scored at 24 h and
72-h post-application.

Cosmetic product (off-white 51 male and female HRIPT. Cream (~ 0.23 g) No dermal irritation or allergic
cream) containing subjects applied for 24 h to upper back contact dermatitis in subjects
0.00554% Mentha Piperita (between the scapulae; area tested.59
(Peppermint) Extract (cm2) not stated). Application
procedure (induction and
challenge) same as reported in
preceding study. Challenge
reactions scored at same
intervals.

Cosmetic product (off-white 26 male and female Maximization test. The test No evidence of contact
cream) containing subjects site (upper outer arm; area not sensitization.60
0.00554% Mentha Piperita stated) pre-treated with 0.25%
(Peppermint) Extract aqueous SLS, applied under
occlusive patch for 24 h.
Product (0.05 ml) then applied,
under occlusive patch, to same
site for 48 h (or for 72 h, if
placed over weekend). Product
application followed by re-
application of SLS patch for 24
h. Sequence repeated for total
of 5 induction exposures.
After 10-day non-treatment
period, new test site (on
opposite arm) pre-treated with
SLS prior to application of
challenge patch. At challenge,
product (0.05 ml) applied for
48 h, under an occlusive patch,
to same site. Reactions scored
at time of patch removal and
48 h later.

100% Mentha Piperita 52 subjects HRIPT (protocol details not Negative results.11
(Peppermint) Leaf Extract included).
(water/ethanol extract)

20% Mentha Piperita 107 subjects (96 women, In HRIPT, product applied Product did not induce
(Peppermint) Leaf Water in 11 men) with no history (0.02 ml) to the back using cumulative irritation or
a face cream of atopy occlusive patch (small Finn sensitization.61
chamber), and 9 induction
applications made over 3-week
period. For 1st, 2nd, 5th, 7th,
and 8th applications, duration
of exposure was 48 ± 4 h.
Duration of exposure was 72 ±
4 h for 3rd, 6th, and 9th
applications. Challenge phase
consisted of single application
to new site and a previously
treated site for 48 ± 4 h.
 Distributed for comment only -- do not cite or quote

Table 6. Dermal Irritation and Sensitization Data

Ingredient Subjects/Cell Type Protocol Results

20% Mentha Piperita 104 male and female In HRIPT, test substance No evidence of irritation or
(Peppermint) Oil subjects applied (0.2 ml) for 24 h to sensitization.56
upper back (between scapulae)
using 3/4" x 3/4" semi-
occlusive patch. Application
repeated 3 times per week for
total of 9 induction
applications. 24-h challenge
patch applied after 2-week
non-treatment period, and
reactions scored at 24 h and 72
h.

20% Mentha Piperita 101 male and female In HRIPT, nine 24-h No evidence of sensitization.57
(Peppermint) Oil subjects applications of test substance
(0.2 ml) made to back using
semi-occlusive patches
(dimensions not stated). 24-h
challenge patch applied after
10- to 15-day non-treatment
period.

Multicenter Studies

Mentha Piperita 73 patients Patients patch tested according Neither irritant nor allergic
(Peppermint) Oil to International Contact reactions reported.62
(concentration not stated) Dermatitis Research Group
(ICDRG) patch test procedures
during 1994 to 1998

Mentha Piperita 13,398 patients in study Patients patch tested during Positive reaction prevalence rate
(Peppermint) Oil (2% in (71 patch tested with years 2009 to 2014 to of 0.53% reported for Mentha
petrolatum) ingredient) determine frequency of Piperita (Peppermint) Oil.64
positive patch test reactions to
essential oils. Study used a
retrospective analysis of patch
test results and relevant
demographical/clinical data
that were collected
electronically by the
US/Canadian North American
Contact Dermatitis Group
(NACDG) and other networks.

Case Reports

Mentha Piperita Male patient with Patch test Allergic positive reaction.65
(Peppermint) Oil (2% in orofacial granulomatosis
petrolatum) mainly affecting lower
lip

Mentha Piperita Female patient with Patch and prick tests Positive patch test reaction, i.e.,
(Peppermint) Oil lichenoid eruption on itching, erythema, and swelling,
(concentration not stated) oral mucosa beginning at day 5; ++ reaction
by day 7. Prick test results
negative.66

Mentha Piperita Male patient with Patch test Strong positive reactions to
(Peppermint) Oil (2% in history of hand eczema LAT patch and to Mentha
petrolatum) and sensitization to Piperita (Peppermint) Oil (2% in
tixocortol pivalate. petrolatum).67
Presented with severe
eczematous contact
dermatitis after repeated
applications of a local
action transcutaneous
(LAT) patch for lumbar
pain containing Mentha
Piperita (Peppermint)
Oil.
 Distributed for comment only -- do not cite or quote

Table 6. Dermal Irritation and Sensitization Data

Ingredient Subjects/Cell Type Protocol Results

Mentha Piperita Female patient with Patch test. Reactions evaluated Positive patch test reaction (+
(Peppermint) Oil (2% in allergic contact at 2, 3 and 7 days according to reaction) observed on days 2 and
petrolatum) dermatitis after ICDRG procedures. 3.68
consuming herbal tea
containing Mentha
Piperita (Peppermint)
Oil.

Mentha Piperita 4 patients with allergic Patch test. Reactions evaluated Positive patch test reactions to
(Peppermint) Oil contact cheilitis (lips and at 48 h and 96 h. lip balm and to Mentha Piperita
(concentration not stated) perioral skin), secondary (Peppermint) Oil.69
to exposure to lip balm
that contained Mentha
Piperita (Peppermint)
Oil

Mentha Piperita Male patient with IgE- Skin prick and prick-to-prick Patient had strongly positive
(Peppermint) Leaf and mediated anaphylaxis to tests prick test reaction to slurry of
Peppermint (concentration peppermint (Mentha peppermint candy and fresh
not stated) piperita) after sucking peppermint leaf. Prick testing of
on peppermint candy. 5 patient with saline slurry of
healthy controls peppermint candy caused wheal
and flare, with largest diameters
of 10 mm and 35 mm
(W10/F25), respectively. Prick-
to-prick test with fresh
peppermint leaf revealed skin
test response of W25/F50. All
prick tests on 5 controls
negative.72

Mentha Piperita Female patient became Skin prick test Positive skin prick test reaction
(Peppermint) Leaf Extract symptomatic with to commercial Mentha Piperita
(concentration not stated) dyspnea when near (Peppermint) Leaf Extract.73
peppermint (Mentha
piperita) scent

Mentha Piperita Male patient with severe Patch test Negative reaction..70
(Peppermint) (concentration cheilitis
not stated)

Mentha Piperita Female patient with Patch test + reaction on days 2 and 4.71
(Peppermint) fragrance recurrent irritant rash
(1:50, 2% in petrolatum) after applying a Mentha
Piperita (Peppermint)
foot spray
 Distributed for comment only -- do not cite or quote

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34. Food and Drug Administration (FDA). Drug products containing certain active ingredients offered over-the-counter (OTC) for
certain uses. 21 CFR 310.545. http://www.accessdata.fda.gov/SCRIPTs/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=310.545.
Last Updated 2016. Date Accessed 2-27-2017.

35. United Sates Environmental Protection Agency (EPA). List of active ingredients eligible for minimum risk pesticide products.
https://www.epa.gov/sites/production/files/2015-12/documents/minrisk-active-ingredients-tolerances-2015-12-15.pdf.
Last Updated 2015. Date Accessed 2-23-2017.

36. European Medicines Agency Committee on Herbal Medicinal Products (HMPC). Community herbal monograph on Mentha x
piperita L., aetheroleum. http://www.ema.europa.eu/docs/en_GB/document_library/Herbal_-
_Community_herbal_monograph/2010/01/WC500059313.pdf. Last Updated 2007. Date Accessed 2-24-2017.

37. Muhammad, F. Wiley J. and Riviere J. E. Influence of some plant extracts on the transdermal absorption and penetration of
marker penetrants. http://dx.doi.org/10.3109/15569527.2016.1147456. Cutaneous and Ocular Toxicology. Last
Updated 2016. Date Accessed 5-16-2016.

38. Nielsen, J. B. Natural oils affect the human skin integrity and the percutaneous penetration of benzoic acid dose-dependently.
Basic Clin Pharmacol Toxicol. 2006;98(6):575-581.

39. Lazutka, J. R., Mierauskiene, J, Slapsyte, G, and Dedonyte, V. Genotoxicity of dill (Anethum graveolens L.), peppermint
(Mentha x piperita L.) and pine (Pinus sylvestris L.) essential oils in human lymphocytes and Drosophila melanogaster.
Food Chem Toxicol. 2001;39(5):485-492.

40. Consumer Product Testing Co. 2017. Bacterial reverse mutation assay (trade name mixture containing 2.5% Mentha Piperita
(Peppermint) Extract).

41. Samarth, R. M. and Kumar A. Mentha piperita (Linn.) leaf extract provides protection against radiation induced chromosomal
damage in bone marrow of mice. Indian Journal of Experimental Biology. 2003;41:229-237.
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42. Samarth, R. M. Panwar M. and Kumar A. Modulatory effects of Mentha piperita on lung tumor incidence, genotoxicity, and
oxidative stress in benzo[a]pyrene-treated Swiss albino mice. Environmental and Molecular Mutagenesis.
2006;47:192-198.

43. Kumar, A. Samarth R. M. Yasmeen S. Sharma A. Sugahara T. Terado T. and Kimura H. Anticancer and radioprotective
potentials of Mentha piperita. BioFactors. 2004;22:87-91.

44. Jain, D. Pathak N. Khan S. Raghuram G. V. Bhargava A. Samarth R. and Mishra P. K. Evaluation of cytotoxicity and
anticarcinogenic potential of Mentha leaf extracts. International Journal of Toxicology. 2011;30(2):225-236.

45. Yi, W. and Wetzstein H. Y. Anti-tumorigenic activity of five culinary and medicinal herbs grown under greenhouse conditions
and their combination effects. J.Sci.Food Agric. 2011;91:1849-1854.

46. Hussain, A. I. Anwar F. Nigam P. S. Ashrafd M. and Gilanif A. H. Seasonal variation in content, chemical composition and
antimicrobial and cytotoxic activities of essential oils from four Mentha species. J.Sci.Food Agric. 2010;90:1827-
1836.

47. Sun, Z. Wang H. Wang J. Zhou L. and Yang P. Chemical composition and anti-inflammatory, cytotoxic, and antioxidant
activities of essential oil from leaves of Mentha piperita grown in China. PLos One. 2014;9(12):e114767

48. Liu, X. Sun Z. L. Jia A. R. Shi Y. P. Li R. H. and Yang P. M. Extraction, preliminary characterization and evaluation of in vitro
antitumor and antioxidant activities of polysaccharides from Mentha piperita. Int.J.Mol.Sci. 2014;15:16302-16319.

49. Liu, Y. Flynn T. J. Ferguson M. S. Hoagland E. M. and Yu L. Effects of dietary phenolics and botanical extracts on
hepatotoxicity-related endpoints in human and rat hepatoma cells and statistical models for prediction of
hepatotoxicity. Food and Chemical Toxicology. 2011;49:1820-1827.

50. Akdogan, M. Kilinc I. Oncu M. Karaoz E. and Delibas N. Investigation of biochemical and histopathological effects of Mentha
piperita L. and Mentha spicata L. on kidney tissue in rats. Human and Experimental Toxicology. 2003;22:213-219.

51. Inoue, T. Sugimoto Y. Masuda H. and Kamei C. Effects of peppermint (Mentha piperita L.) extracts on experimental allergic
rhinitis in rats. Biol.Pharm.Bull. 2001;24(1):92-95.

52. Oh, J. Y., Park, MA, and Kim, YC. Peppermint Oil Promotes Hair Growth without Toxic Signs. Toxicol Res. 2014;30(4):297-
304.

53. Derma Consult GmbH. 2015. Human patch test of a lipstick containing 0.2961% Mentha Piperita (Peppermint) Leaf Extract.

54. Peritesco. Ocular and cutaneous acceptability study of a cleansing gel (containing 50% Mentha Piperita (Peppermint) Leaf
Water) during 4 weeks under ophthalmological and dermatollgical control. Unpublished data submitted by the Personal
Care Products Council on 3/1/2016. 2008. pp.1-42.

55. 4-Front Research. Single patch test of a cleansing gel containing 50% Mentha Piperita (Peppermint) Leaf water. Unpublished
data submitted by the Personal Care Products Council on 3/1/2016. 2007. pp.1-23.

56. Consumer Product Testing Company. Repeated insult patch test on 20% Mentha Piperita (Peppermint) Oil. Unpublished data
submitted by the Research Institue for Fragrance Materials (RIFM) on 3/13/2017. 2001. pp.1-16.

57. TKL Research, Inc. Repeated insult patch test on 20% Mentha Piperita (Peppermint) Oil. Unpublished data submitted by the
Research Institute for Fragrance Materials (RIFM ) on 3/13/2017. 2002. pp.1-20.

58. Consumer Product Testing Co. 2017. Repeated insult patch of a trade name mixture containing 2.5% Mentha Piperita
(Peppermint) Extract.

59. Consumer Product Testing Co. 2012. Executive summary of a repeated insult patch test of cosmetic product (off-white cream
containing 0.00554% Mentha Piperita (Peppermint) Extract).

60. Ivy Laboratories (KGL Inc). 2011. Executive summary of a human maximization test of an off-white cream containing 0.00554%
Mentha Piperita (Peppermint) Extract.

61. Institut d'Expertise Clinique. Sensitization and cutaneous compatability study (face cream containing 20% Mentha Piperita
(Peppermint) Leaf Water). Unpublished data submitted by the Personal Care Products Council on 3/1/2016. 2010.
pp.1-11.
 Distributed for comment only -- do not cite or quote

62. Kanerva, L., Rantanen, T, Aalto-Korte, K, Estlander, T, Hannuksela, M, Harvima, RJ, Hasan, T, Horsmanheimo, M, Jolanki, R,
Kalimo, K, Lahti, A, Lammintausta, K, Lauerma, A, Niinimki - Section of Dermatology, FIoOHHF, Turjanmaa, K, and
Vuorela, AM. A multicenter study of patch test reactions with dental screening series. Am J Contact Dermat.
2001;12(2):83-87.

63. Consumer Product Testing Co. 2017. The MatTek Corporation sub-Draize mildness testing (MTT ET-50) using the EpiOcular™
tissue model in vitro toxicity testing system (trade name mixture containing 2.5% Mentha Piperita (Peppermint)
Extract).

64. Warshaw, E. M. Zug K. A. Belsito D. V. Fowler J. F. Jr. DeKoven J. G. Sasseville D. Maibach H. I. Mathias C. G. T. DeLeo V.
A. Taylor J. S. Fransway A. F. Marks J. G. Jr. Pratt M. D. Zirwas M. J. Geier J. and Uter W. Positive patch-test
reactions to essential oils in consecutive patients: Results from North America and Central Europe. Dermatitis.
2017;28(4):246-252.

65. Lewis, F. M., Shah, M, and Gawkrodger, DJ. Contact sensitivity to food additives can cause oral and perioral symptoms. Contact
Dermatitis. 1995;33(6):429-430.

66. Fleming, C. J. and Forsyth A. Short communications. D5 patch test reactions to menthol and peppermint oil. Contact Dermatitis.
1998;38:337

67. Foti, C., Conserva, A, Antelmi, A, Lospalluti, L, and Angelini, G. Contact dermatitis from peppermint and menthol in a local
action transcutaneous patch. Contact Dermatitis. 2003;49(6):312-313.

68. Vermaat, H., van, MT, Rustemeyer, T, Bruynzeel, DP, and Kirtschig, G. Vulval allergic contact dermatitis due to peppermint oil
in herbal tea. Contact Dermatitis. 2008;58(6):364-365.

69. Tran, A., Pratt, M, and DeKoven, J. Acute allergic contact dermatitis of the lips from peppermint oil in a lip balm. Dermatitis.
2010;21(2):111-115.

70. Guin, J. D. Rosemary cheilitis: one to remember. Contact Dermatitis. 2001;45:63

71. Kalavala, M., Hughes, TM, Goodwin, RG, Anstey, AV, and Stone, NM. Allergic contact dermatitis to peppermint foot spray.
Contact Dermatitis. 2007;57(1):57-58.

72. Bayat, R. and Borici-Mazi R. A case of anaphylaxis to peppermint. Allergy, Asthma and Clinical Immunology. 2014;10(6):1-3.

73. Szema, A. M. and Barnett T. Allergic reaction to mint leads to asthma. Allergy Rhinol. 2011;2:43-45.

74. Amjadia, M. A. Mojabb F. and Kamranpoura S. B. Th effect of peppermint oil on symptomatic treatment of pruritus in pregnant
women. Iranian Journal of Pharmaceutical Research. 2012;11(4):1073-1077.

75. Kearns, G. L., Chumpitazi, BP, Abdel-Rahman, SM, Garg, U, and Shulman, RJ. Systemic exposure to menthol following
administration of peppermint oil to paediatric patients. BMJ. 2015;5(8):1-6.

76. Tisserand, R. and Young R. Essential Oil Safety. A Guide for Health Care Professionals. 2nd ed. New York: Elsevier, 2014.

77. Wallace, R. B. Gryzlak B. M. Zimmerman M. B. and Nisly N. L. Application of FDA adverse event report data to the
surveillance of dietary botanical supplements. The Annals of Pharmacotherapy. 2008;42:653-660.

78. International Agency for Research on Cancer (IARC). IARC Monograph on Pulegone.
https://monographs.iarc.fr/ENG/Monographs/vol108/mono108-05.pdf. Last Updated 2017.

79. Da Rocha, M. S. Dodmane P. R. Arnold L. L. Pennington K. L. Anwar M. M. Adams B. R. Taylor S. V. Wermes C. Adams T. B.
and Cohen S. M. Mode of action of pulegone on the urinary bladder of F344 rats. Toxicol.Sci. 2012;128(1):1-8.

80. Ale, S. O. Hostynek J. J. and Maibach H. L. Menthol: A review of its sensitization potential. Exog.Dermatol. 2002;1:74-80.
 Distributed for comment only -- do not cite or quote

2017 FDA VCRP Data

Mentha Piperita (Peppermint) Oil
01B - Baby Lotions, Oils, Powders, and Creams 1
02A - Bath Oils, Tablets, and Salts 24
02B - Bubble Baths 2
02D - Other Bath Preparations 8
03G - Other Eye Makeup Preparations 4
04B - Perfumes 1
04E - Other Fragrance Preparation 17
05A - Hair Conditioner 43
05B - Hair Spray (aerosol fixatives) 1
05F - Shampoos (non-coloring) 57
05G - Tonics, Dressings, and Other Hair Grooming Aids 31
05H - Wave Sets 1
05I - Other Hair Preparations 15
07E - Lipstick 88
07I - Other Makeup Preparations 34
08B - Cuticle Softeners 2
08E - Nail Polish and Enamel 2
08G - Other Manicuring Preparations 3
09A - Dentifrices 34
09B - Mouthwashes and Breath Fresheners 19
09C - Other Oral Hygiene Products 73
10A - Bath Soaps and Detergents 43
10B - Deodorants (underarm) 2
10E - Other Personal Cleanliness Products 26
11A - Aftershave Lotion 3
11B - Beard Softeners 2
11D - Preshave Lotions (all types) 2
11E - Shaving Cream 10
11G - Other Shaving Preparation Products 2
12A - Cleansing 33
12C - Face and Neck (exc shave) 30
12D - Body and Hand (exc shave) 45
12E - Foot Powders and Sprays 9
12F - Moisturizing 56
12G - Night 1
12H - Paste Masks (mud packs) 17
12I - Skin Fresheners 13
12J - Other Skin Care Preps 73
Total 827

Mentha Piperita (Peppermint) Leaf (No Posting)

Mentha Piperita (Peppermint) Leaf Extract

02D - Other Bath Preparations 1
03D - Eye Lotion 4
 Distributed for comment only -- do not cite or quote

04C - Powders (dusting and talcum, excluding aftershave

talc) 1
04E - Other Fragrance Preparation 4
05A - Hair Conditioner 23
05B - Hair Spray (aerosol fixatives) 1
05F - Shampoos (non-coloring) 18
05G - Tonics, Dressings, and Other Hair Grooming Aids 3
05I - Other Hair Preparations 5
07B - Face Powders 2
07C - Foundations 2
07D - Leg and Body Paints 1
07E - Lipstick 5
07I - Other Makeup Preparations 2
08F - Nail Polish and Enamel Removers 1
08G - Other Manicuring Preparations 1
09A - Dentifrices 1
09B - Mouthwashes and Breath Fresheners 1
10A - Bath Soaps and Detergents 6
10E - Other Personal Cleanliness Products 6
11A - Aftershave Lotion 3
11G - Other Shaving Preparation Products 1
12A - Cleansing 14
12C - Face and Neck (exc shave) 24
12D - Body and Hand (exc shave) 5
12E - Foot Powders and Sprays 1
12F - Moisturizing 17
12G - Night 2
12H - Paste Masks (mud packs) 8
12I - Skin Fresheners 10
12J - Other Skin Care Preps 24
13A - Suntan Gels, Creams, and Liquids 1
Total 198

Mentha Piperita (Peppermint) Leaf Water

03G - Other Eye Makeup Preparations 2
10A - Bath Soaps and Detergents 1
10B - Deodorants (underarm) 1
11G - Other Shaving Preparation Products 1
12A - Cleansing 3
12C - Face and Neck (exc shave) 1
12F - Moisturizing 3
12H - Paste Masks (mud packs) 1
12I - Skin Fresheners 1
12J - Other Skin Care Preps 1
Total 15

Mentha Piperita (Peppermint) Extract

 Distributed for comment only -- do not cite or quote

02A - Bath Oils, Tablets, and Salts 1

05A - Hair Conditioner 1
05E - Rinses (non-coloring) 1
05F - Shampoos (non-coloring) 1
05G - Tonics, Dressings, and Other Hair Grooming Aids 1
07E - Lipstick 2
07I - Other Makeup Preparations 2
09B - Mouthwashes and Breath Fresheners 2
10A - Bath Soaps and Detergents 2
10B - Deodorants (underarm) 1
10E - Other Personal Cleanliness Products 3
11A - Aftershave Lotion 1
11E - Shaving Cream 3
12A - Cleansing 4
12C - Face and Neck (exc shave) 10
12D - Body and Hand (exc shave) 2
12E - Foot Powders and Sprays 3
12F - Moisturizing 17
12H - Paste Masks (mud packs) 10
12I - Skin Fresheners 1
12J - Other Skin Care Preps 3
Total 71
Distributed for comment only -- do not cite or quote
Distributed for comment only -- do not cite or quote
Distributed for comment only -- do not cite or quote
Distributed for comment only -- do not cite or quote
 WORKPLACE
ENVIRONMENTAL
EXPOSURE
LEVEL
Menthol(2014)
I. IDENTIFICATION
Statement on similar isomer activity relationship: The different
Chemical Name: Menthol menthol isomers are considered similar for the purposes of
Chemical Name toxicological assessments.(2–4)
Synonyms
(CAS #)(1)
2-Isopropyl-5-methylcyclohexanol; 5- Molecular Formula: C10H20O
Methyl-2-(1-methylethyl)cyclohexanol; Structural Formula:
DL-Menthol Raw Menthol; Menthyl alcohol;
(1490-04-6) Cyclohexanol, 5-methyl-2-(1-
methylethyl)-; DL-Menthol; 5-Methyl-2-
(1-methylethyl) cyclohexanol;
Racementhol; D/L-Menthol Racemate;
Menthol; Racemic menthol; (+-)-
Menthol; (1R,2S,5R)-Menthol; 5-
Methyl-2-(1-methylethyl) cyclohexanol,
(1alpha,2beta, 5alpha)-; 2-isopropyl-5-
methylcyclohexanol; Hexahydrothymol;
Menthacamphor; Menthol natural;
DL-Menthol Menthol natural, brazilian; Menthol (2,4,5–16)
Synthetic racemic; Menthomenthol; Peppermint II. CHEMICAL AND PHYSICAL PROPERTIES
(89-78-; Former camphor; Racementhol; Physical State: Crystals or granules
15356-70-4) Racementholum; Racementol; p- Molecular Weight: 156.27
Menthan-3-ol; rac-Menthol; (+-)- Conversion Factors 1 ppm = 6.4 mg/m3; 1 mg/m3 = 0.16 ppm
(1R*,3R*,4S*)-Menthol; Cyclohexanol, Melting Point: 41-43 oC (106-109 oF) Isomer not specified. DL-
5-methyl-2-(1-methylethyl)-, Menthol exists in two polymorphs melting at 28 oC and 38
(1R,2S,5R)-rel-; Cyclohexanol, 5- o
C, respectively
methyl-2-(1-methylethyl)-, (1alpha, Freezing point: 27 to 28 oC, rising on prolonged stirring to 30
o
2beta,5alpha)-; Menthol; Menthol, cis- C to 32 oC. Isomer not specified
1,3,trans-1,4-; d,l-Menthol; dl-Menthol Boiling Point: 212 oC (414 oF) Isomer not specified
(-)-Menthol; Levomenthol; (l)-Mentho; Density/Specific Gravity: 0.890 g/cm3 Isomer not specified;
(-)-Menthyl alcohol; (-)-trans-p- 0.895 g/cm3 at 20 °C (CAS# 1490-04-6)
Menthan-cis-ol; (1R)-(-)-Menthol; (1R- Vapor Pressure: 8.5 Pa (0.064 mm Hg) at 25 °C (L-menthol,
(1-alpha,2-beta,5-alpha))-5-Methyl-2-(1- Isomer not specified); 30 Pa (0.975 mm Hg) at 55 °C D/L
methylethyl) cyclohexanol; D-(-)- menthol
Menthol; Levomenthol; Saturated Vapor Concentration: 84 ppm (538 mg/m3) at 22-25
Levomentholum; Menthol, (1R,3R,4S)- °C (calculated; = Vapor Pressure X 1315); 132 ppm (845
L-Menthol Natural (-)-; Menthol, l-; U.S.P. Menthol; l-(-)- mg/m3) at 55 °C (calculated; = Vapor Pressure X 1315)
or Synthetic Menthol; l-Menthol; l-Menthol (natural); Octanol/Water Partition Coefficient: log Kow = 3.2–3.4 CAS-
(2216-51-5) (-)-(1R,3R,4S)-Menthol; (-)-Menthol; No. 1490-04-6, 2216-51-5, 15356-70-4, Isomer not
Cyclohexanol, 5-methyl-2-(1- specified
methylethyl)-, (1R,2S,5R)-; Solubilities:
Cyclohexanol, 5-methyl-2-(1- Water solubility: 456 mg/l @ 25 °C (Isomer not specified);
methylethyl)-, (1R,3R,4S)-; 431–508 mg/l at 20 - 25°C flask method (CAS# 1490-04-6,
Cyclohexanol, 5-methyl-2-(1- 2216-51-5, 15356-70-4, Isomer not specified);
methylethyl)-, (1R- Slightly soluble in water; very soluble in alcohol,
(1alpha,2beta,5alpha))- chloroform, ether, petroleum ether acetone, benzene,

Copyright 2014
OARS – 2300 Montana Avenue, Suite 409, Cincinnati, OH 45211
 2

organic solvents; freely soluble in glacial acetic acid, liquid Menthol

petrolatum Species Route LD50 or LC50
Isomer
Odor Description and Threshold: Peppermint odor;
Threshold odor concentration, detection 0.14–0.26 ppm, 0.9 940(3)
- 1.7 mg/m3 2602(4)
Oral
Threshold odor concentration 0.002–11.6 mg/ m3
Threshold odor concentration, recognition 0.33 ppm, 2.1 2900(3)
mg/m3 Rat 3180(4)
Taste: Peppermint
Flammability Limits: Not available 5289 mg/m³ (29)
Inhalation
Flash Point: > 100°C (closed cup; isomer not specified; purity > (4h, aerosol)
99.7 %) DL- IM 10,000(16)
Autoignition Temperature: Not available
IP 750(3)
Other Properties:
Specific optical rotation: +49.2 degree/D (alcohol, 5%, D- Oral 3100(25)
menthol) Mouse
SubQ 14000-16000(3)
UV: 635 (Sadtler Research Laboratories Spectral
Collection) Rabbit IP 2000(3)
NMR: 410
Hazardous Reactivities & Incompatibilities: Oral 1500 – 1600(3)
Cat
Incompatible with phenol; beta-naphthol; resorcinol or IP 1500 – 1600(3)
thymol in trituration; potassium permanganate; chromium
trioxide; pyrogallol, butylchloral hydrate, camphor, 2426 – 2615*(4)
phenol, chloral hydrate, exalgine Oral 3300(3)
Hazardous Decomposition:
When heated to decomposition it emits acrid smoke and Rat 3400(25)
irritating fumes.
IP 710(3)
III. USES AND VOLUMES SubQ 1000 – 2500(3)
Menthol is an alcohol produced from mint oils or prepared 4380(3)
synthetically. Peppermint oil contains about 35–60 % menthol L- Oral
Mouse 2600(3)
(menthone (15–30 %), menthylacetate (4–14 %), and small
amounts of cineole and other terpenes).(17) Menthol is widely IP 2000(3)
used in consumer products as well as other uses. Product types
SubQ 5000-6000(3)
were listed as antipruritics, dermatologic agents, approved as a
Group II pesticide, flavoring substance, flavoring agents, Guinea Pig IP 4000(3)
denaturants, fragrance ingredients, used in cosmetics as
denaturant, masking, refreshing, soothing agents, FDA direct IP 800 – 1000(3)
Cat
food additive, paints and lacquers, adhesives, metal care Oral 800 – 1000(4)
products, cleaning products, shoe- and leather-care products,
disinfectants, solvents, cosmetics, odor improvers, repellents L-/DL- Rabbit Dermal ≥ 5000(4)
and animal care products, farming of cattle and other animals, Rat Oral 3180(16)
manufacture of foodstuffs, manufacture of pharmaceutical and
medicinal chemicals and of cleaning products. Concentrations INS Oral > 6000(4)
for oral products range from 0.001–2%; dermal products range Mouse
Dermal 34500(28)
from 0.001–6%; and inhaled products from 0.1–0.45%.(4,18–24)
*Females; h – hours; INS – isomer not specified; SubQ –
Subcutaneous; IP – Intraperitoneal; IM – Intramuscular
IV. TOXICOLOGY DATA
Death occurred 1-3 days after dosing. Clinical signs were
A. Acute Toxicity and Irritancy reported as narcotic status and depressed activity but no data
1. Lethality Data were available on exposure level at which the effects were
observed.(3,4) At sublethal doses of L-menthol in mice, lethargy
Menthol
Species Route LD50 or LC50 was reported.(25) In one study with L- menthol, a lower LD50 of
Isomer 940 mg/kg was observed.(4) In this study, a severe irritation of
D- Rat Oral 2046(4) the mucosal lining of the stomach and intestine was reported.
Other investigators have not reported such effects.
DL- Rat Oral 940(3)

Copyright OARS For personal use only. Do not distribute.

 3

2. Eye Irritation condition associated with increased airway secretions/mucous

membrane irritation. CNS-related effects were rapidly
Concentrations of 29 to 64% of L-, D- or DL-menthol in
reversible. Bradypnea and labored breathing patterns were
diethylphthalate, undiluted or 71% menthol liquid were tested
observed through post exposure day 10. The respirability of the
for eye irritation in the same institute and by the same OECD
aerosol was adequate to achieve the objective of study, i.e., the
Guideline 405 protocol. The vehicle was tested in the opposite
average mass median aerodynamic diameter was 3.1μm ± 1.8
eye of the animals, and showed no irritating properties. Mean
(geometric standard deviation).(29)
scores 24, 48, and 72 h after the various menthol isomer
treatments indicated concentration dependent reactions of
Several acute irritation studies were available. In mice, 16
cornea and conjunctiva. In all cases, there was no reaction
ppm for 15 min caused mild irritation in the upper respiratory
observed in the iris. After treatment with menthol liquid (100%
tract as determined by changes in respiratory function.(28)
and 71%), slight redness of the conjunctiva was seen on day 7
Sprague-Dawley rats were exposed to 0.13 mg/m3 (0.02
in 1/4 and 2/4 animals, respectively. For the undiluted menthol
ppm) menthol as part of a cold preparation vapor for 4 or 8
liquid it was shown that these effects were completely
hours in a whole body chamber. No differences in bacterial
reversible within 14 days.(4) In another rabbit study, instillation
clearance were observed between control and treated rats.(29)
of 1% or 5% solution in an unknown vehicle or undiluted
The sensory irritation potential of menthol was evaluated in 30-
menthol (of unknown purity), were reported to result in eye
minute exposures of Swiss-Webster mice to seven menthol
injury (grade 9 on a scale of maximum 10). No details were
concentrations ranging from 18 to 31 ppm (115 to 198 mg/m3).
available on the number of animals or the nature of the effects
Periocular wetness was observed in several animals 24 hours
seen after menthol treatment.(26) Additionally in a Draize test,
following exposure to concentrations of 22 ppm (140 mg/m3)
concentrations of 10% to 60% DL-menthol were applied to the
and above, and mortalities were recorded among the 20 and 30
eyes of eight animals with four eyes rinsed after one minute and
ppm (140 and 191 mg/m3) exposure groups. The airborne
four without rinsing. In this test, menthol was not considered
concentration resulting in a 50% decrease in respiratory rate
irritating.(4)
(RD50) in anesthetized mice was determined to be 45 ppm (288
3. Skin Absorption mg/m3).(30,31)
No data other than lethality are available.
B. Subacute Toxicity
4. Skin Irritation
Groups of six male mice were dosed with 2000, 2500, 3200,
Concentrations ranging from 1% to undiluted L-, D-, DL- 4000, or 5000 mg/kg L-menthol by gavage for five days and
menthol or menthol liquid in diethylphthalate were tested for monitored for 14 days. The LD50 was 2600 mg/kg.(3)
skin irritation in the same institute and by the same OECD
Guideline 404 protocol. All undiluted isomers were irritating to Ten male and 10 female Wistar rats per group were gavaged for
the skin. No skin reactions were observed for D-menthol and 28 days with 200, 400 or 800 mg/kg/day L-menthol. The study
menthol liquid at the 5% dilution, and for L- and DL-menthol at was conducted mainly according to OECD guideline 407.
the 1% dilution.(4) Animals were checked for clinical signs and mortality twice
daily. Body weight, food and water consumption were recorded
5. Sensitization
weekly. At necropsy, kidneys, adrenals, heart, brain, liver and
Sensitization studies were conducted on L-menthol. In an the stomach (with contents) weights were taken. Hematology
OECD 406 Buehler test, 0.5 ml of a 25 % solution was applied consisting of hemoglobin, PCV, total erythrocyte count, total
in an occlusive dressing to guinea pig skin for both the white blood cells, white blood cell differential count, and
induction and the challenge phases.(4) A local lymph node assay reticulocyte counts; biochemistry consisting of glucose,
was performed according to the protocol of Kimber and creatinine, urea, and liver enzyme activities; and urine for
Weisenberger.(4) In both studies, L-menthol was negative. In a presence of blood, ketones, glucose and proteins were
modified Draize test, a positive result was only obtained when examined. The organs were prepared for histological
four 0.1 ml induction injections of 0.1% L-menthol and two examination as described in OECD guideline 407 with the
challenge periods consisting of a 0.1 ml intradermal injection of exception of the full histopathology examinations of urinary
the 0.1% solution and a topical application of a 10% solution bladder and prostate. There was no effect of treatment on body
were used.(4) A local lymph node assay was performed with weight gain, food consumption or clinical chemistry. There was
concentrations up to 50% DL-menthol and was negative.(27) significantly increased water consumption and an increased
number of neutrophilic granulocytes at the highest dose level;
6. Inhalation Toxicity however, for both effects, neither the magnitude nor the sex was
In one study, bradypnea, labored breathing pattern, dyspnea, reported. In males at all doses and females at ≥ 400 mg/kg/day,
motility reduced, atony, tremor, high-legged gait, staggering absolute and relative liver weights were significantly increased
gait, movements uncoordinated, piloerection, ungroom hair (no information on magnitude and incidence of this findings is
coat, nasal discharge (serous, red), stridor, breathing sounds, given in the publication). In all treated males and females,
apathy, narcosis, prostration, miosis, hypothermia, decreased histopathological examination revealed vacuolization of
reflexes, and transient decrease in body weights were observed. hepatocytes (male and female combined totals were as follows.
The authors stated that the effects were suggestive of a narcotic Controls, 0/20; 200 mg/kg, 4/20; 400 mg/kg, 5/17; 800 mg/kg,

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 4

4/19). The liver effects were not dose-related and were excretion of glucuronides, water, or electrolytes, or interference
interpreted by the authors as a possible adaptation process. with central nervous system reactions to stimulants were
Since no information is available as to the magnitude and the observed. The NOAEL was considered 200 mg/kg/day.(3)
incidence of increased liver weights in the various exposure
groups, the relevance of this finding is questionable and a Menthol solutions of 1% and 5% were applied daily to the nasal
NOAEL or a LOAEL could not be deduced from this study.(4) mucous membrane of a rabbit for nine months. Animals were
examined daily for amount and type of nasal discharge, general
C. Subchronic Toxicity state of activity and body weight. The histopathology of the
nasal mucosa was examined from the posterior or
Male and female B6C3F1 mice were treated with DL-menthol ethmoturbinate. Degenerative changes were observed with
in the diet for 13 weeks with a post exposure period of one
destructive changes throughout all layers of the nasal membrane
week. The dose groups of 10 males and 10 females each
after treatment with 5% menthol solution.(4)
included the vehicle, 930, 1870, 3750, 7500 or 15000 ppm
(male mice: 0, 243, 488, 978, 1956 or 3913 mg/kg/day; female
Twelve male and female Sherman Rats per group were exposed
mice: 290, 595, 1193, 2386 or 4773 mg/kg/day, respectively). for 6.75 hours daily to 0.087, 0.148 or 0.259 ppm (according to
Mortality was examined daily. Clinical signs including 0.57, 0.96 or 1.68 mg/m³, respectively) L-menthol in whole
appearance and behavior, body weight and food consumption
body inhalation chambers for 71–79 days. Two control groups
were recorded weekly. At the end of the exposure period,
of 12 animals were utilized; one under identical conditions
organs were examined according to OECD guideline 408 except
except without menthol exposure and the other group remained
the aorta, peripheral nerve and spinal cord were not examined. in their cages throughout the study. Although the authors
Histopathology was conducted for the controls, 7500 and 15000 attempted to measure the menthol concentrations in the gas
ppm concentration groups. No differences were found between
phase, there was no adequate analytical method available.
treated and controls with respect to clinical signs, mortality,
Therefore the exposure concentrations are given as weight of
time to death, food consumption, gross pathology or
menthol vaporized divided by the volume of air circulated.
histopathology. For males and females, treatment at the highest
Clinical signs, mortality, food consumption and water
dose of 15000 ppm, 3913 mg/kg/day and 4773 mg/kg/day, consumption were examined daily. Body weights were
respectively, decreased body weight gain by 5 -10 % compared examined twice a week. Organ weights, hematology, and
to controls. Findings described as spontaneous lesions included
microscopic examinations of the eye, turbinates, nasopharynx,
kidney (interstitial nephritis, nonsuppurative pyelitis) and early
trachea, lungs, and skin, sections of liver, spleen, kidney, heart,
spontaneous respiratory disease lesions (peribronchial or
testes, ovaries, intestine and skeletal muscle were conducted.
perivascular lymphoid hyperplasia, lung congestion). The
Mortality and time to death, body weight gain, food and water
NOAELs for male and female mice were 7500 ppm, consumption, hematology, organ weights, and gross pathology
corresponding to 1956 mg/kg/day and 2386 mg/kg/day, were not different from controls. During daily clinical sign
respectively, based on reduced body weight gain.(4)
examinations, transient erythema of the conjunctiva was
observed, which then disappeared shortly after the animals were
Male and female Fischer 344 rats were treated with DL-menthol
returned to their cages. Upon histopathological examination of
in the diet for 13 weeks with a post exposure period of one
the lung in the highest dose group, respiratory tract effects were
week. The dose groups of 10 males and 10 females each
observed that included tracheitis, pneumonitis, pulmonary
included vehicle, 930, 1870, 3750, 7500 or 15000 ppm (males:
congestion and severe congestion to pneumonitis, which was
0, 59, 114, 231, 472 or 937 mg/kg/day; females: 0, 67, 142, 285,
suggestive of irritation. A NOAEL or LOAEL could not be
521 or 998 mg/kg/day, respectively). Mortality was examined
assigned due to invalid analytical methods for measurement of
daily. Clinical signs, including appearance and behavior, body
the menthol air concentrations.(4)
weight and food consumption, were recorded weekly. At the
end of the exposure period, organs were examined according to A 13-week rat inhalation study comparing 200, 600 or 1200
OECD guideline 408 except that the aorta, peripheral nerve and
mg/m3 smoke particulates of which 2% was L-menthol (4, 12 or
spinal cord were not examined. Histopathology was conducted
24 mg/m3; 0.6, 1.9 or 3.8 ppm, respectively) to non-menthol
for the control, 7500 and 15000 ppm concentration groups. No
containing cigarette smoke, demonstrated that menthol did not
differences were found between treated and controls with
increase the respiratory tract histopathological lesions observed
respect to clinical signs, mortality, time to death, food after inhalation of cigarette smoke alone. No further conclusions
consumption, gross pathology or histopathology. Findings of could be drawn on the toxicity of menthol since a menthol-only
questionable relevance at the high dose included minimal (32)
increase in the severity of spontaneous interstitial nephritis. The treatment group was not included in the study.
NOAEL for male and female rats was 15000 ppm,
corresponding to 937 mg/kg/day and 998 mg/kg/day, D. Chronic Toxicity/Carcinogenicity
respectively, the highest dose tested.(4) B6C3F1 mice were treated with 0, 2000 or 4000 ppm
(approximately 334 or 667 mg/kg/day) DL-menthol in the feed
In a dietary study in male and female rats exposed to 0, 100 or for 103 weeks. Clinical signs and mortality were checked twice
200 mg/kg/day L-menthol or DL-menthol for 5.5 weeks, with daily. Body weights and food consumption was determined
limited end points examined, no adverse effects on weight gain, every two weeks. At the end of the one-week post-exposure

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 5

period, organs were preserved and examined according to for chromosomal aberrations or sister chromatid exchanges in
OECD Guideline 451. There were no treatment related effects Chinese hamster fibroblast cells, Chinese hamster ovarian cells,
on survival, clinical signs of toxicity, food consumption or human embryonic lung cells, human peripheral lymphocytes or
gross pathology. A slight decrease in body weight gain human TK6 cells blood lymphocytes. There were no significant
estimated at less than 10% was observed in treated groups. increases in polyploidy or number of aberrations.(37,43–48)
There was a slight increase in the incidence of hepatocellular Several comet assays were conducted in Chinese hamster ovary
carcinomas in males. However, it was not statistically K5 cells.(49) Other systems were used to examine menthol
significant and was within the range of historical controls. mutagenicity: Bacillus subtilis (L-menthol), E. coli WP2
Overall, the conclusion was that there was no increased uvrA(trp-), anaphase chromosome aberrations test in human
incidence of neoplasms compared to controls. The NOAEL for tissue culture cells (fibroblasts), carcinoma prediction assay in
males and females was 667 mg/kg/day.(33) C3H/10T1/2 cells carrying bovine papilloma virus DNA (DL-
menthol), umu DC-lacZ genes in S. typhimurium strain
Fischer 344 rats were treated with 0, 3750 or 7500 ppm TA1535/pSK1002 (DL-menthol). The results were
(approximately 188 or 375 mg/kg/day) DL-menthol in the feed negative.(3,41,50–52) D-Menthol was negative in the comet assay
for 103 weeks. Clinical signs and mortality were checked twice using either V79 hamster cells or human lymphocytes at
daily. Body weights and food consumption was determined concentration up to 2 mM with and without activation.(4) In an
every two weeks. At the end of the one-week post-exposure alkaline elution assay to detect DNA damage in primary rat
period, organs were preserved and examined according to hepatocytes from 0.1 mM up to cytotoxic concentrations of 1.3
OECD Guideline 451. There were no treatment related effects mM, D-menthol was negative.(53) Only in a DNA repair test in
on survival, clinical signs of toxicity, food consumption or bacillus subtilis M 45 (rec-) and H17 (rec+) at up to 10 mg/disk
gross pathology. However, there was a slight decrease in body was menthol considered positive.(41) At cytotoxic concentration
weight gain estimated at less than 10% for the low and high in an in vitro alkaline elution/rat hepatocyte assay, DL-menthol
dosed males and the low dose females and approximately 14% was positive; however, when tested at non cytotoxic
for the high dose females. A common finding in aged rats, concentrations of 0.1–1.3 mM, DL-menthol was not considered
increased chronic renal inflammation was found in treated male genotoxic.(53)
rats compared to controls (29/49, 41/50 and 41/50,
respectively). Treated female rats showed a decrease in Several in vivo tests were conducted with DL-menthol. Two
mammary gland fibroadenomas compared to controls (20/50, oral cytogenetic assays in male rats were conducted using a
20/49 and 7/43, respectively). Overall, the conclusion was that single dose of 1.45, 14.5, 145, 500 or 3000 mg/kg; or 5 daily
there was no increased incidence of neoplasms compared to doses of 1.45, 14.5, 145 or 1150 mg/kg. Five animals per dose
controls. The overall NOAEL was 188 mg/kg/day, which was and time point were injected with colcemid 2 hours prior to
based on females showing decreased body weight gain with a termination at 6 hours (all animals in 5 day study), 24 hours or
difference greater than 10% compared to control rats.(33) 48 hours. Bone marrow analysis for chromosome aberrations
did not show a difference between vehicle and treated
E. Developmental / Reproductive Toxicity animals.(3) Two dominant lethal assays in male rats were
conducted in which ten animals were either given a single dose
Four oral developmental toxicity studies were conducted to
of 1.45, 14.5, 145, 500 or 3000 mg/kg; or 5 daily doses of 1.45,
examine the effect of oral DL-menthol exposure during 14.5, 145 or 1150 mg/kg followed by mating with 2 females per
gestation in the mouse, rat, hamster, or rabbit. Mouse, rat, week for eight or seven weeks, respectively. Fertility index,
hamster and rabbit maternal and fetal NOELs were 185, 218,
preimplantation loss and lethal effects on the embryos were
405 and 425 mg/kg, respectively, the highest doses tested. No
examined. There were no significant differences between
effects on survival of dams, body weight of dams or average
treated and control animals.(3) In a host mediated assay, in
fetal weight were observed. No fetotoxicity, abnormalities/ which mice were treated with either a single dose of 1.45, 14.5,
malformations, or skeletal findings or soft tissue abnormalities 145, 500 or 3000 mg/kg; or 5 daily doses of 1.45, 14.5, 145 or
compared to control group were observed.(4) But the positive
1150 mg/kg menthol and using the indicator organisms
control fetuses did experience altered vertebrae.
Salmonella typhimurium (his TA 1530 or G-46) or
Saccharomyces cerevisiae (D-3), three hours after injection
F. Genotoxicity/Mutagenicity with the indicator organism, animals were killed and yeast or
Multiple in vitro and in vivo studies were conducted on the bacterial species were collected and recombinants were
isomers/racemates of menthol. In the Ames study with L or DL- counted. In three of the assays, the results were negative. In one
isomers using bacterial strains of Salmonella typhimurium assay after 5 daily doses, there was a slightly enhanced
(TA92, TA94, TA97, TA97a, TA98, TA100, TA102, TA1530, recombinant frequency at all doses. In a second test, at the
TA1535, TA1537, TA2637 and G-46), and Escherichia coli highest dose of 1150 mg/kg using Saccharomyces cerevisiae
(WP2 uvrA) with and without metabolic activation at up to (D-3), the result was negative.(3) In vivo mouse micronucleus
cytotoxic concentrations, which ranged from of 500 to 1000 after three intraperitoneal doses of 250, 500 or 1000 mg/kg DL-
µg/plate depending on the strain; menthol was negative.(34–41) In menthol; or an in vivo comet assay (alkaline single cell gel
a mouse lymphoma mutation assay in L5178Y mouse electrophoresis) in ddy mice after oral gavage administration of
lymphoma cells, DL-menthol was negative.(42) L- and/or DL- 2000 mg/kg in olive oil, DL-menthol was negative.(54–57) A
menthol with and without metabolic activation was examined replicative DNA synthesis test was conducted in mouse or rat

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 6

hepatocytes from male B6C3F1 mice or F344 rats, respectively, 4. Excretion

that had received a single oral dose at 1000 or 2000 mg/kg, the
In rats, menthol is primarily metabolized to the glucuronide
maximum tolerated dose (MTD) in corn oil. After 24, 39 or 48
conjugate and eliminated in the urine or feces.(58–60) In one study
hours, hepatocytes were prepared and radiolabelled thymidine
in rats, 470 mg/kg of [3-3H]-menthol (unspecified isomer) was
incorporation was measured in vitro. DL-menthol was positive
administered orally, 52% of the radioactivity was found in the
in mice at both the dose levels at 24 hours. In the high dose
urine, 4.5% and 3.5% were found in the feces and ileum 17
group, there was also a significant increase in cell viability at 24
hours after dosing.(58) After a single dose of 500 mg/kg of
hours. In rats, DL-menthol was positive at 24 hours in the low
radiolabeled L-menthol to male uncannulated Fischer 344 rats,
dose group and at 39 hours in the high dose group.(55,56)
71% of the dose was found in urine and feces 48 hours later.
After the first 24 hours, 45% of the dose was recovered with
Although 3 studies indicated a positive result with respect to
18% and 27% of the total radiolabel in the urine and feces,
inducing chromosomal aberrations, these positive responses
respectively. At 48 hours, a similar percentage of the total dose
appear to be due to cytotoxic concentrations of menthol. The
was recovered in the urine and a lower concentration of 7.3% in
weight of evidence indicates that menthol is not genotoxic.(4)
the feces. The same treatment was given to bile duct-cannulated
rats where the total recovery of radiolabeled material in the
G. Metabolism and Pharmacokinetics
urine and bile was 74%, the majority (67%) being recovered in
1. Absorption the bile. The major metabolite found in the bile was menthol
glucuronide and various oxidation products were found in the
The isomers L-, DL- and the unspecified menthol isomer
urine.(59)
mixture appear to be well absorbed orally in rats with reports of
≥ 63% to ≥ 74%(58–60) and in rabbits with reports of ≥ 86% to Other species have been examined for elimination of menthol.
90%.(61–63) In sheep fed with L-menthol, L-menthyl glucuronide was
detected in the urine and within 24 hours after consumption the
Dermal absorption is lower than oral absorption.(63) To assess in excretion was considered almost complete.(65) In the urine of
vitro skin penetration, radiolabeled, neat L-menthol was applied rabbits fed 1 g/kg of DL-menthol or L-menthol, DL-menthol
to rat skin using flow-through diffusion cells under either and L-menthol glucuronides were found in similar amounts of
occluded or unoccluded conditions. The receptor fluid was 59% and 48% of the dose, respectively.(62,66) In rabbits fed 3 g
collected at 48 hours and analyzed, demonstrating that 3% and of menthol, 86% was eliminated by glucuronidation, even when
1% was absorbed under occluded conditions compared to this maximum toxic dose was given.(61) The last of 24 daily
(64)
unoccluded skin, respectively. doses of 2 g menthol, 90% was excreted as menthol glucuronide
within 6 hours. The glucuronide was a minor urinary excretion
In a mouse model using racemic menthol, the menthol upper product in dogs, suggesting that other metabolic routes would
respiratory tract uptake efficiency into the systemic circulation be more important in this species.(62) (Williams, 1938). In the
was measured to be 90.3% ± 1.1 and 66.3% ± 2.8 at the inspired dog, 5% of a 5 g oral dose of menthol was excreted in the urine
concentration of 1.3 ppm and 18.3 ppm menthol for 15 minutes, as the glucuronide conjugate.(61)
respectively.(28)
2. Distribution H. Other

A single oral dose of 470 mg/kg [3-3H]-menthol (unspecified Three Wistar rats were exposed to L-menthol for 4 weeks by
isomer) was administered to rats and after 17 hours, 52% of whole body inhalation at a concentration of 1.6 x10-13 M
radioactivity was found in urine, 4.5% in feces, 3.5% in ileum, (0.000025 mg/m3; 3.9 x10-6 ppm) continuously. Control animals
2.1% in fat, 0.8% in liver, 0.31% in serum, 0.2% in kidney, and were exposed to filtered fresh air only. The rats were sacrificed
< 0.1% in brain and testes.(58) after 4 weeks of exposure and the mitral cells of olfactory bulbs
were examined. L-Menthol exposure produced selective
3. Metabolism degeneration of the mitral cells in various sections of the
Yamaguchi and colleagues have investigated the metabolism of olfactory bulb.(67) The authors state that “degeneration in this
menthol and menthol glucuronide in rats.(59) Menthol context in no way implies cell death” and the cells in the
glucuronide is formed in the liver and passes into the bile, heavily degenerated zones behave normally.
where the molecule is eliminated or enters the enterohepatic
circulation. Various oxidation reactions may occur upon Menthol has pharmacological activity with the effect being
subsequent passages through the liver. The oxidation products considered a chemesthesis effect, which is defined as
of menthol include para-menthane-3,8-diol, para-menthane- “sensations that arise when chemical compounds activate
3,9-diol, and 3,8-dihydroxy- para-menthane-7-carboxylic receptor mechanisms for other senses, usually those involved in
acid.(59,60) The oxidation metabolites, a primary alcohol, a triol, pain, touch, and thermal perception in the eye, nose, mouth and
and hydroxy acids have also been identified.(59) In situ nasal throat.”(68) Cooling of the upper airway, which stimulates
metabolism of menthol occurs, as evidenced by the decreased specific cold receptors and inhibits laryngeal mechanoreceptors,
absorption efficiency in CYP450 inhibitor metyrapone- reduces respiratory activity in un-anesthetized humans and
pretreated mice.(28) anesthetized animals. More recent investigations have provided
evidence for menthol to increase cough thresholds(69,70) but only

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when it is administered as vapor to the upper airway.(71) ankyrin 1 (TRPA1) -/- mice. This suggests that the metabolite
Menthol is an agonist of transient receptor potential melastatin- likely acts through TRPA1. Thus, the direct stimulation of
8 (TRPM8) receptor, which is a cationic ion channel rat dorsal sensory nerves (e.g., the sensory irritant response) and the
root ganglion(72) involved in detection of normal cooling- counter irritation are likely due to differing molecules (e.g.,
sensation in mammals.(73) In studies, cold air or warm air and L- metabolite vs. parent menthol).(28)
menthol in anesthetized new born dogs or guinea pigs (390 ng
L-menthol/ml for 10 seconds duration with an airflow of 30 The subcutaneous administration of menthol produced
ml/s) greatly reduced ventilation.(74,75) In an investigation of ambulation in mice. From experiments conducted with
neurophysiological responses in individual fibers of the lingual dopamine agonists and a monoamine-depleting compound, the
and chorda tympani nerves and in single cortical neurons, in the authors suggested that dopamine is involved in the abilities of
rat, lingual fibers and a different category of cortical units (Type menthol to promote ambulation in mice.(81)
II) were extremely sensitive to menthol exposure.(76)
Several studies have shown that menthol-containing
An analgesic effect as determined by a significant reduction in formulations function to enhance dermal penetration.(82,83)
pain during functional tasks was observed after application of a Synchrotron X-ray diffraction was employed to evaluate the
3.5% gel to osteoarthritic individuals.(77) Also in animals, an effect of ethanol and L-menthol on lipid arrangements in the
analgesic effect was observed when application of 40% menthol stratum corneum of hairless rats. It was shown that L-menthol
to the contralateral rat hind paw tended to reduce responses to was dispersed through the stratum corneum, intruded mainly
cooling and noxious heat.(78) In patch clamp and tetrodotoxin into hexagonal hydrocarbon chain packing, and disrupted the
mediated Na+ channel blockade in vitro and in mice, the role for regular organization of these structures.(84)
Na+ channel blockade in DRG neurons was demonstrated in the
efficacy of menthol as topical analgesic compound.(79) Some studies have investigated the involvement of menthol in
metabolic processes. A single oral dose of 470 mg/kg [3-3H]-
A study demonstrated the roll of TRPM8 lacrimation after low menthol (unspecified isomer) administered to rats resulted in
concentration instillations to the cornea. Tear measurements 70% inhibition of HMG-CoA reductase activity 17 hours after
were made using a cotton thread in TRPM8 wild type and treatment, which returned to normal activity by 41 hours.(60)
knockout mice after application of menthol (0.05-50 mM) to the Male Wistar rats exposed for two weeks to dietary 0.5% or 1%
cornea. In additional studies, nocifensive responses (eye menthol caused an increase in serum cholesterol and serum
swiping and lid closure) were quantified following cornea triglycerides levels in the high-dose group, but no effect on apo
menthol application. Trigeminal ganglion electrophysiologic A-1 lipids, an indicator of high-density lipoprotein status or
single unit recordings were performed in rats to determine the body weights. Liver weight was slightly increased.(85)
effect of low and high concentrations of menthol on corneal
cool cells. At low concentrations, menthol increased tear MacDougall reported that that L-menthol and synthetic
production in TRPM8 wild type and heterozygous animals, but congeners inhibit the microsomal oxidation of nicotine to
had no effect in TRPM8 knockout mice, while nocifensive cotinine and the P450 2A6-mediated 7-hydroxylation of
responses remained unaffected. At the highest concentration, coumarin in vitro.(86)
menthol (50 mM) increased tearing and nocifensive responses
in TRPM8 wild type and knockout animals. This study indicates V. HUMAN USE AND EXPERIENCE
low concentrations of menthol (0.1 mM) responses were via the
TRPM8, yet a high concentration of menthol increased tearing The pharmacokinetics of menthol has been described in
and nocifensive responses were via a separate mechanism.(80) humans. The oral absorption of menthol ranged between 10 and
90%. Menthol is primarily metabolized to the glucuronide
Willis and colleagues examined the interaction of menthol and conjugate and excreted almost completely in the urine within 12
cigarette smoke in the lung conducted studies. Menthol acts as a to 24 hours.(61,87–92) In a crossover placebo-controlled study,
broad-spectrum counterirritant, diminishing the chemosensory twelve subjects received three 100 mg L-menthol capsules, a
responses to inhaled irritants. In a mouse model using racemic placebo capsule, and 10 mg menthol in mint candy or mint tea.
or L-menthol, the effect of menthol pre-treatment on acrolein Plasma and urine levels of menthol and conjugated menthol
irritation was investigated. It was shown that 16 ppm menthol (glucuronide), cardiovascular measurements, and subjective
attenuated a 2 ppm acrolein induced breathing pattern change; effects were measured. Only the menthol glucuronide could be
however, 4 ppm menthol did not diminish the effect. This measured in plasma or urine. The plasma half-life of menthol
attenuation effect of menthol was significantly diminished in glucuronide averaged 56.2 minutes and 42.6 minutes under the
animals treated with AMTB, a TRPM8 antagonist. The menthol capsule and mint candy/mint tea conditions,
counterirritant properties of menthol appear to be due to parent respectively. Urinary recovery of menthol as the glucuronide
menthol itself rather than a CYP450 metabolite, as counter averaged 45.6 and 56.6% for menthol capsule and mint
irritation was fully apparent in P450 inhibitor, metyrapone- candy/tea, respectively.(93) In patients with liver disease,
pretreated mice. Conversely, the CYP450 metabolite of menthol alcohol-induced cirrhosis or steatosis, doses of 2 g menthol
appears to be responsible for sensory irritation as evidenced by were given and menthol glucuronide was determined. The mean
inhibition of menthol sensory irritation by metyrapone and the excretion of menthol glucuronide was slightly lower than
absence of sensory irritation in transient receptor potential healthy subjects and the authors conclude that patients with

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 8

liver disease retain a significant capacity to metabolize tilted the head forward to hold the solution in the anterior of the
menthol.(3) Glucose-6-phosphate-dehydrogenase-deficiency in oral cavity, and then agitated it gently with the tongue. They
newborn babies may result in development of severe jaundice were prompted to expectorate at 10 seconds, then instructed to
after menthol administration due to the inability of the neonates keep the mouth closed to prevent evaporative cooling. At 15
to conjugate menthol.(94) seconds (5 seconds after expectoration) and at 45 seconds,
subjects were asked to rate the intensity of irritation and
When considering pharmacokinetics after dermal applications, coolness in the mouth. Mean ratings of sensations of
an unspecified amount of menthol containing ointment was irritation produced by a high concentration of racemic menthol
applied to the skin and urine samples were collected. The (0.3% w/v) decreased significantly over repeated exposures,
excretion of menthol was stated to be slower after dermal even when the time between stimuli was as long as 5 minutes.
absorption than after oral administration. Additionally, the urine This shows menthol is capable of producing desensitization to
of an untreated person living in the same room as a patient sensory irritation in the oral cavity.(99)
rubbed with a menthol-containing ointment was analyzed and
menthol was detected. The authors concluded that a large Dermal sensitization of menthol has been investigated in
percentage of menthol absorbed after dermal application was controlled studies and described in case reports. In a
inhaled.(63) In another study, a number of commercial patches maximization test with 8% DL-menthol in petrolatum (5520
(2, 4 or 8) containing 37.44 mg menthol (isomer not specified) µg/cm2) performed with 25 volunteers, there were no positive
were applied to the skin of 8 subjects (4 male, 4 female) for 8 reactions(100). In 9 human patch tests using DL- or L- menthol
hours. For the 4 and 8-patch groups, the average maximum with 6227 patients with dermatological disease, a low incidence
plasma concentrations (Cmax +/- SD) were 19.0 +/- 5.4 and of positive responses, 0.3 to 6.1% positive reactions, were
31.9 +/- 8.8 ng/mL, respectively. The 2-patch group had demonstrated.(4,101–109) Based on the negative results obtained in
measurable but low plasma concentrations. The harmonic mean several animal and human studies with L- and DL-menthol, the
terminal half-live was 4.7 +/- 1.6 hours. The absolute dermal widespread use of menthol in consumer dermal contact products
bioavailability could not be determined in this study; however, and the low number of skin reactions reported in dermal
the authors concluded that there appears to be relatively low compromised patients or case reports, dermal sensitization is of
systemic exposure even when an unrealistically large number of low concern for menthol.
patches were applied for an extended period.(95)
A solution of 0.5% or 0.2% L-menthol in petrolatum or 0.1% L-
In the literature, there have been a number of studies and case menthol in saline was applied to the nasal passages of 16
reports that provided a summary of the potential adverse effects subjects three times per day at 2 day intervals. The 0.5%
after menthol exposure by various routes. The usual human oral concentration was considered irritating, the 0.2% concentration
dose is 60-120 mg menthol per person.(3) The maximum doses was considered almost non-irritating to non-irritating and the
tested in humans in pharmacokinetic studies were 180 mg(87) 0.1% concentration was considered non-irritating to the nasal
and 1000 mg.(61) It is reported that about 20 mg/kg led to a mild and mucous membranes.(110)
abdominal discomfort.(91) Three volunteers were exposed orally
with 8000 to 9000 mg (approximately 120 mg/kg) of an Menthol provides a cooling sensation to the skin and respiratory
unspecified isomer of menthol which resulted in cold burning tract that appears to be pharmacologically mediated. Menthol is
sensation in mouth, throat and esophagus, a cold sensation on an agonist of transient receptor potential melastatin-8 (TRPM8),
the mucous membranes of the nose, on the skin of the hands and a cationic ion channel that is involved in detection of normal
feet, and fatigue.(96) Abdominal pain, convulsions, nausea, cooling-sensation in mammals.(73) The psychophysical effects of
vomiting, vertigo, ataxia, drowsiness and coma have been TRPM8 activation in humans by application of 40% menthol
reported after ingestion of high doses of menthol.(97,98) Over- solution for 20 minutes on the forearm was examined in 10
dosage with menthol (isomer not specified) over an extended volunteers. All subjects experienced pain from the 40% menthol
period of time has resulted in gastrointestinal distress, ataxia, application described as burning, pulling, freezing, cutting,
stupor, convulsions and blood dyscrasias.(4) The WHO estimates tingling; hot, cold, spreading, dull or taut. Quantitative sensory
the human oral lethal dose to be approximately 50–500 testing and laser Doppler imaging was performed before and
mg/kg.(3) after exposure. Menthol produced no axon reflex reaction and
resulted in cold hyperalgesia.(111)
In a crossover placebo-controlled study, twelve subjects
received 3 exposures of 100 mg L-menthol capsule, or a Another study investigated the ability to perceive gradual
placebo capsule and evaluated for cardiovascular and subjective increases in skin temperature on the vermilion border of the lip
effects. Following menthol capsule ingestion, the decrease in after application of 0.2 or 2.0% L-menthol in mineral oil. Supra-
heart rate was less than the decrease after placebo threshold sensations of warmth could be significantly attenuated
administration.(93) and the threshold for warmth was increased significantly
whereas the threshold for heat pain was unchanged by exposure
In another study examining sensory irritation, subjects received to menthol.(112) In a sensory perception test, 0.5% menthol in
ten menthol solutions at one of two concentrations. After the ultrapure water was applied to the nasolabial fold of 58 adult
subject rinsed three times with distilled water, 10-ml samples (19 male and 39 female) volunteers for 2.5, 5 or 8 minutes. The
were presented at 1-min intervals. Subjects sipped the sample, volunteers completed a questionnaire during the treatment to

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 9

provide information on the type and intensity of any sensory treatment time of 15 minutes. Two patients in the menthol
effect. Menthol at 0.5% elicited stinging and cooling sensations. group withdrew from the study because of an uncomfortable
A significant response (sensory score of 3 or more) after sensation in the upper airway. As measured by expiratory flow
exposure to 0.5% menthol was observed in 22/58 subjects. rates, vital capacity, and forced expiratory volume, menthol
However, after screening these 22 volunteers for cooling improved airway hyper-responsiveness at doses as low as 20 mg
sensation only 9 were classified as being sensitive to 0.5% per day.(117)
menthol.(113)
The literature is sparse with respect to air concentrations of
Menthol was demonstrated to affect ventilation in humans in menthol in the workplace and how these relate to adverse
several studies as was shown in animals. Total nasal resistance effects. Thymol was used as an indicator for menthol in the
to airflow was measured in 31 subjects before and after a five- Bayer menthol manufacturing plant in 1990-91. Thymol has
minute exposure to 0.2 mg/L (200 mg/m3; 31 ppm) menthol chemical properties similar to menthol, e.g., a melting point of
vapor. Menthol inhalation had no consistent effect on nasal about 50 °C and a boiling point of 233 °C. The results of the
resistance but the majority of subjects reported an increased thymol measurements were < 0.5 mg/m³ and < 0.8 mg/m³ in the
sensation of nasal airflow and a cooling effect of menthol. The Bayer menthol manufacturing factory.(4) An investigation of
results indicate that menthol stimulates cold receptors in the occupational exposures to menthol vapors during the
nasal mucosa to create an increased sensation of airflow.(114,115) manufacture of mentholated Sucret’s throat lozenges was
conducted in response to employee complaints of respiratory
In young (18 to 26 years) or elderly (over 65 years) healthy and ocular irritation. Effects described by production workers
volunteers, 9–10 per group; 0.21, 0.42, 0.85, 1.70, 3.39, 6.78 or included local irritation of the eyes, nasal passages, throat and
13.56 ppm (1.3, 2.7, 5.4, 11, 22, 43 or 87 mg/m3) menthol was larynx. Non-smokers complained of runny nose, redness and
inhaled through the nostrils by a Dravniecks Dynamic Dilution watering of the eyes and physical exams found inflammatory
Binary Scale Olfactometer. An odor threshold was measured changes in the nasal mucosa, vocal cords and throats. Seven
using the up-down staircase method. Intensity and pleasantness participants with suspected nasal polyps may represent an
were measured by magnitude estimation. The average threshold excess over the expected occurrence in the normal population.
for the elderly participants (approximately 0.7 ppm) was Pre and post exposure pulmonary function testing showed
significantly higher than for young participants (0.26 ppm). The significant decreases in forced vital capacity (FVC) and 1-
median slope of the intensity function was steeper by a factor of second forced expiratory volume (FEV1) for non-smoking
two for younger adults. A 10-fold increase in menthol individuals. The total population showed a decrease in FVC and
concentration produced a four-fold increase in perceived increases in FEF 25–75%, FEF 75–85% and MEF 75%. Air
intensity for young adults and a two-fold increase in perceived sampling indicated that menthol was present in the air of
intensity for elderly persons. The younger persons had a steeper packaging and wrapping areas, which ranged from non-
average pleasantness function and found menthol less pleasant detectable to 2.3 mg/m3 (0.4 ppm). Although exposure
with repeated exposure; however, menthol concentrations concentration comparisons were not evaluated, it was reported
corresponding to perceived unpleasantness were not that the menthol in the air in the cooling and candy rooms was
provided.(12) noticeably higher and more irritating. Air concentrations in
these rooms ranged from 1.9 to 39.4 mg/m3 (0.8 to 6.2 ppm)
A study was conducted to examine olfactory and chemosensory with a mean and median of 12.7 and 11.8 mg/m3 (2.0 and 1.8
threshold. It was found that the absolute odor threshold was ppm), respectively. Typical symptoms described while working
lower than the chemosensory threshold. Absolute detection in in these areas included immediate stinging, watering and tearing
both the nasal and oral cavities was based on olfaction and not of the eyes upon entering the room followed by moderate
stinging, cooling or taste. The individual threshold irritation of the nasal passages and throat. One 15 minute air
concentration was 5.00 x 10-5 M to 5.10 x 10-2 M (three orders sample in the breathing zone of a NIOSH industrial hygienist by
of magnitude variation) in PEG200 (relative headspace the candy machine was 39.4 mg/m3 (6.2 ppm) and the
concentration ranging from 0.002 µg/L to 11.600 µg/L), with an symptoms described by this individual during that period were
exception of two data points (3.00 x 10-3 M, 1.20 x 10-2 M), and immediate stinging and tearing of the eyes, soreness and
with an overall geometric mean threshold concentration of 3.42 dryness in the tonsil area of the throat, a cooling irritation of the
mM.(13) nose, watery nasal discharge, periodic (non-productive)
coughing, and tingling sensation in the face and arms. Cold
Twenty-five employees exposed to an unspecified menthol sweating occurred for about five minutes after leaving the candy
isomer (concentration not specified) were examined room.(118)
olfactometrically. A control group was also examined which
consisted of 25 employees working in the same plant, but not Several incident reports and case reports were identified.
exposed to menthol. The examination showed a general Inhalation of high doses of menthol was reported to cause
diminution of smell acuity on an odor identification task.(116) adverse CNS effects. A woman developed insomnia, unsteady
The airway hyper-responsiveness of 23 human subjects with gait, mental confusion, depression, vomiting, and cramp in the
chronic mild asthma was tested by use of a nebulizer containing legs after smoking 80 mentholated cigarettes per day for 3
10 mg menthol twice a day for four weeks. An estimate of the months.(119) In another report, a 13-year old boy inhaled an
air concentration was 32 mg/m3 (5 ppm) assuming a nebulizer

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estimated 200 mg menthol in a menthol and olbas oil mixture the two are considered to have same toxicity. Menthol has low
and experienced similar symptoms.(120) acute and chronic toxicity potential in mammals. The chemical
is neither genotoxic, carcinogenic nor a reproductive or
A case of asthma due to menthol exposure was reported in a 40- developmental toxicant. It is not a dermal sensitizer, and is not
year-old woman with no history of asthma or any other allergy, absorbed through the skin in toxicologically significant
presenting with dyspnea, wheezing and nasal symptoms after quantities. However, menthol was irritating to the eyes, skin and
using menthol containing toothpaste and candies. Menthol was respiratory tract.
confirmed as the causative agent by positive skin tests and
bronchial challenge.(121) Using the NOAEL (188 mg/kg, females) from a chronic oral rat
study with the lowest effect being decreased weight gain and
In several epidemiology studies, the effect of smoking applying uncertainty factors for intra and interspecies variability
mentholated cigarettes as a risk factor in various cancers was would result in a WEEL greater than that proposed below. The
investigated. Current cases of cigarette smokers with 588 male most sensitive effect considered as the point of departure for the
lung cancer cases and 914 male controls, and 456 female lung WEEL was on the lung, including irritancy, lacrimation, and
cancer cases and 410 female controls were investigated. The receptor mediated cooling pharmacological responses.
prevalence of menthol usage did not differ between cases and
controls of either sex. For specific histological types of lung The derivation of the WEEL considered the weight of evidence
cancer (squamous cell carcinoma, small cell carcinoma, large regarding the dose-response from several human and animal
cell carcinoma and adenocarcinoma) there was no indication of studies, as there was no clear NOEL in a standard toxicity study
an association with menthol usage.(122) Carpenter and colleagues by the inhalation route. It is acknowledged that an acclimation
concluded that lung-cancer risk from smoking mentholated to the irritation and lacrimation occurs in the workplace and that
cigarettes resembled the risk from smoking non-mentholated there is a lack of chronic inhalation dose response data. In a
cigarettes from examining a population of 337 incidents of lung subchronic inhalation study in rats,(128) there was a clear NOEL
cancer compared to 478 controls.(123) An additional cohort study for respiratory histopathological effects with no systemic effects
investigating the use of mentholated cigarettes and lung cancer observed. Even though the air concentration could not be
in men and women was conducted. The study population verified, the NOEL indicates that this effect is a dose-
consisted of 11761 members (5771 men, 3990 women). The dependent, threshold-type irritant effect reasonably expected
relative risk of lung cancer associated with mentholation after repeated exposure. Three reports indicate a WEEL of 1
compared with non-mentholated cigarettes was 1.45 in men and ppm would prevent respiratory irritation. Application of modest
0.75 in women. The authors’ conclusion was that there was an uncertainty factors to either 1) the acute mouse pharmacology
increased risk of lung cancer associated with mentholated study, which identified a 4-ppm no effect level for a counter-
cigarette use in male smokers but not in female smokers.(124) In irritancy effect(30); or 2) the mouse RD50 of 45 ppm multiplied
another study, it was investigated whether smoking mentholated by 0.03,(30,31,129) both support a WEEL of 1 ppm (6.4 mg/m3). In
cigarettes increased the risk of cancer of the oral cavity and addition, 3) the NIOSH report(118) described menthol air
pharynx. One hundred and ninety-four males and 82 females concentrations in the workplace areas associated with some
were test subjects and 845 male and 411 female controls were complaints of irritation as 0.8 to 6.2 ppm with a median range
part of the study. From this analysis, menthol was not a risk of 1.8 and 2.0 ppm. This indicates a WEEL of 1 ppm would be
factor for cancer and it was concluded that the use of appropriate. It is reasonable to consider that shorter exposures
mentholated cigarettes is unlikely to be an important to higher concentrations of 3 ppm (19.2 mg/m3) would not
independent factor in oropharyngeal cancer.(125) Next, the cause noticeable irritation. However, this STEL may not protect
relationship of menthol cigarette smoking and esophageal all naive individuals of severe lacrimation upon entering an area
cancer was investigated. Data from a large hospital-based case- containing this air concentration of menthol. Based on the
control study was used. There was no change in the cancer risk weight of evidence, a Workplace Environmental Exposure
for males ever-smoking menthol versus those never smoking Level of 1 ppm (6.4 mg/m3) and a 15 minute Short Term
menthol cigarettes. For women, however, there was an Exposure Limit of 3 ppm (19.2 mg/m3) are assigned for
increased risk. The authors stated that because of the limitations menthol.
of the study the issue of menthol cigarette smoking and
esophageal cancer could not be resolved.(126) The epidemiology VII. RECOMMENDED WEEL
data overall suggest that smoking mentholated cigarettes does
not increase cancer or other disease risk above that already 8-hr Time-Weighted Average (TWA): 1 ppm (6.4 mg/m3)
present from smoking non-mentholated cigarettes.(127)
15 min Short Term Exposure Limit (STEL): 3 ppm (19.2
mg/m3)
VI. RATIONALE
Menthol is a liquid with a high vapor pressure and a minty odor No additional hazard notations are assigned.
with a low threshold. This chemical is used widely in the
consumer products and food industries where low
concentrations are added to products with direct oral and dermal
exposure. It was found that D- and L-isomers or a mixture of

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