Final Report on the Safety Assessment of Calendula

OfŽcinalis Extract and Calendula OfŽcinalis1

INTRODUCTION
Calendula OfŽ cinalis Extract is an extract of the  owers of The safety of Calendula OfŽ cinalis Extract and Calendula
Calendula ofŽ cinalis, the common marigold, whereas Calendula OfŽ cinalis used in almost 200 cosmetic formulations, ranging
OfŽ cinalis is described as plant material derived from the  ow-
from skin and eye cosmetics through hair products and soaps,
ers of C. ofŽ cinalis. Techniques for preparing Calendula OfŽ ci-
nalis Extract include gentle disintegration in soybean oil. Propy- is reviewed in this report. Both Calendula OfŽ cinalis Extract
lene glycol and butylene glycol extractions were also reported. and Calendula OfŽ cinalis are obtained from the calendula,
Components of these ingredients are variously reported to include Calendula ofŽ cinalis, and serve as biological additives
sugars, carotenoids, phenolic acids, sterols, saponins,  avonoids, (Wenninger and McEwen 1997).
resins, sterins, quinones, mucilages, vitamins, polyprenylquinones,
and essential oils. Calendula OfŽ cinalis Extract is reported to be
used in almost 200 cosmetic formulations, over a wide range of
CHEMISTRY
product categories. There are no reported uses of Calendula Of-
Ž cinalis. Acute toxicity studies in rats and mice indicate that the DeŽnition
extract is relatively nontoxic. Animal tests showed at most min-
imal skin irritation, and no sensitization or phototoxicity. Min- Calendula OfŽ cinalis Extract (CAS No. 84776-23-8) is an
imal ocular irritation was seen with one formulation and no extract of the  owers of the calendula, C. ofŽ cinalis (Wenninger
irritation with others. Six saponins isolated from C. ofŽ cinalis  ow- and McEwen 1997). It is also known as Calendula Extract;
ers were not mutagenic in an Ames test, and a tea derived from Extract of Calendula; Extract of Calendula OfŽ cinalis; and
C. ofŽ cinalis was not genotoxic in Drosophila melanogaster. No car-
Marigold Extract.
cinogenicity or reproductive and developmental toxicity data were
available. Clinical testing of cosmetic formulations containing the Calendula OfŽ cinalis is a plant material derived from the
extract elicited little irritation or sensitization. Absent any basis  owers of C. ofŽ cinalis (Wenninger and McEwen 1997). It
for concluding that data on one member of a botanical ingredi- is also known as Calendula and Calendula Powder. The plant
ent group can be extrapolated to another in a group, or to the Calendula is also known as marigold, garden marigold, pot
same ingredient extracted differently, these data were not consid-
marigold, Marybud, holigold (Fleischner 1985), holligold, and
ered sufŽ cient to assess the safety of these ingredients. Additional
data needs include current concentration of use data; function in gold-bloom (Budavari 1989).
cosmetics; ultraviolet (UV) absorption data; if absorption occurs
in the UVA or UVB range, photosensitization data are needed;
gross pathology and histopathology in skin and other major organ Physical and Chemical Properties
systems associated with repeated dermal exposures; dermal repro-
A mixture of Calendula OfŽ cinalis Extract (1% – 5%), soy-
ductive/developmental toxicity data; inhalation toxicity data, espe-
cially addressing the concentration, amount delivered, and particle bean (Glycine Soja) oil (>50%), and tocopherol (<0.1%) is a
size; and genotoxicity testing in a mammalian system; if positive, reddish-yellow oily liquid with an aromatic herbal odor (Chemis-
a 2-year dermal carcinogenicity assay performed using National ches Laboratorium Dr. Kurt Richter GmbH 1996). It is soluble in
Toxicology Program (NTP) methods is needed. Until these data fats and oils, has a refractive index (nD 20± C) of 1.474 to 1.475,
are available, it is concluded that the available data are insuf-
density of 0.918 to 0.922 g/ml, and an acid value of <1.0. A mix-
Ž cient to support the safety of these ingredients in cosmetic
formulations. ture of Calendula OfŽ cinalis Extract (10% – 25%) and propylene
glycol (>75%) is a clear, brown liquid with a faint herbal odor
(Grau Aromatics GmbH & Co. 1998). It is soluble in water,
has a refractive index of 1.425 to 1.445 (at 20± C), density of
1.035 to 1.055 (at 20± C), and a pH value of 5.5 to 6.5. A mix-
Received 7 January 2001; accepted 21 March 2001. ture of Calendula OfŽ cinalis Extract, butylene glycol, and water
1
Reviewed by the Cosmetic Ingredient Review Expert Panel. (percentages not speciŽ ed) is a yellowish-brown, transparent
Monice Zondlo Fiume, former ScientiŽ c Analyst/Report Management
Coordinator, prepared this report. Address correspondence to Direc- liquid with a bitter taste (Ichimaru Pharcos Co., Ltd. 1994). It
tor, Cosmetic Ingredient Review, 1101 17th Street, NW, Suite 310, has a speciŽ c gravity (d20/20) of 1.01 to 1.05 and a pH of 5.0
Washington, DC 20036, USA. to 6.0.

International Journal of Toxicology, 20(Suppl. 2):13 – 20, 2001

Copyright °c 2001 Cosmetic Ingredient Review
1091-5818/01 $12.00 + .00 13
14 COSMETIC INGREDIENT REVIEW

Manufacture and Production A mixture of Calendula OfŽ cinalis Extract (1% – 5%), soy-
A mixture containing Calendula OfŽ cinalis Extract (1% – bean (Glycine Soja) oil (>50%), and tocopherol (<0.1%) is
5%), soybean (Glycine Soja) oil (>50%), and tocopherol identiŽ ed using the “total of quality control data” (Chemisches
(<0.1%) is characterized as a fatty oil extract of calendula blos- Laboratorium Dr. Kurt Richter GmbH 1996).
soms; the fatty oil used is soybean oil (Chemisches Laborato-
rium Dr. Kurt Richter GmbH 1996). The calendula blossoms are
Ultraviolet Absorption
“gently disintegrated and extracted with stabilized soybean oil.”
Published data on the ultraviolet absorption of Calendula Of-
The mixture is then obtained by Ž ltration.
Ž cinalis Extract and Calendula OfŽ cinalis were not found.
A mixture of Calendula OfŽ cinalis Extract (10% – 25%) and
propylene glycol (>75%) is prepared by extracting calendula
blossoms with 1,2-propylene glycol; the ratio of extract to botan- Impurities
ical is 5:1 (Grau Aromatics GmbH & Co. 1998). A preservative, A mixture of Calendula OfŽ cinalis Extract, butylene glycol,
0.6% phenonip (phenoxyethanol, methylparaben, butylparaben, and water contains ·10 ppm heavy metals and ·1 ppm arsenic
ethylparaben, and propylparaben), is used. (Ichimaru Pharcos Co., Ltd. 1994).
A mixture of Calendula OfŽ cinalis Extract, butylene glycol,
and water (percentages not speciŽ ed) is prepared by extracting
calendula  owers with 1,3-butylene glycol (Ichimaru Pharcos USE
Co. Ltd. 1994).
Cosmetic
Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis are
Composition reported to function as biological additives (Wenninger and
The blossoms of C. ofŽ cinalis contain carotenoid pigments, McEwen 1997). The product formulation data submitted to the
a saponin, a “bitter principle,” and calendulin (Fleischner 1985). Food and Drug Administration (FDA) in 1998 reported that Cal-
Vidal-Ollivier et al. (1990) reported that six saponins were de- endula OfŽ cinalis Extract was used in 178 cosmetic formula-
termined in six cultural varieties of C. ofŽ cinalis, and that the tions, 177 used under the name Calendula Extract and 1 under
concentration of the saponin was dependent upon the variety and the named Calendula Fluid Extract, and that Calendula OfŽ ci-
the date of harvest. Calendula is also composed of volatile oil, nalis was not used (FDA 1998) (Table 1).
caledin (Budavari 1989), and  avonol-3-O-glycosides (Vidal- Concentration of use values are no longer reported to the
Ollivier et al. 1991; Pietta et al. 1992), and C. ofŽ cinalis con- FDA by the cosmetics industry (FDA 1992). Data submitted by
tains acylated steryl glucosides that are composed of C12 – C22 the cosmetics industry reported that one company uses Calen-
fatty acids (Wojciechowski and Zimowski 1975). Extract of dula OfŽ cinalis Extract at concentrations of <0.5% in a styling
C. ofŽ cinalis in propylene glycol contained sugars, carotenoids, gel, a shampoo, and a cream rinse; another company uses it at
 avonoids, and essential oils components and an extract in iso- a concentration of 0.2 weight % (CTFA 1998); and a supplier
propyl myristate contained carotenoids, phenolic acids, sterols, states that a mixture of Calendula OfŽ cinalis Extract (10% – 25%)
and essential oil components (Góra et al. 1980). and propylene glycol (>75%) is used at 1% – 10% in cosmetic
A supplier of a mixture containing Calendula OfŽ cinalis Ex- products (Grau Aromatics, GmbH & Co. 1998). Product formu-
tract and propylene glycol stated that the plant is composed of lation data submitted to the FDA in 1984 stated that Calendula
essential oil, carotenoids,  avonoids, triterpenic alcohols, or- OfŽ cinalis Extract was used in 24 cosmetic formulations, 12 at
ganic acids and esters, sterins, quinones, mucilages, saponins, concentrations of ·10% and 12 at unknown concentrations, and
resins, vitamins, polyprenylquinones , and bitter substances that Calendula OfŽ cinalis was used in 3 cosmetic formulations
(Grau Aromatics GmbH & Co. 1998). A supplier of a mix- at concentrations of <5% (FDA 1984) (Table 2).
ture containing Calendula OfŽ cinalis Extract and butylene gly-
col and water stated that the main elements of the plant are
International
 avonoid, saponine, and amino acid (Ichimaru Pharcos Co., Ltd.
1994). Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis, as
Calendula Extract, are listed in the Japanese Comprehensive
Licensing Standards of Cosmetics by Category (CLS) (Rempe
Analytical Methods and Santucci 1997). Calendula Extract which conforms to the
High-performanc e liquid chromatography (HPLC) (Vidal- speciŽ cations of the Japanese Cosmetic Ingredients Codex
Ollivier et al. 1991), reversed-phase HPLC, and micellular elec- (JCIC) has precedent for use without restriction in all CLS cate-
trokinetic capillary chromatography have been used to analyze gories. Calendula OfŽ cinalis, as Calendula Powder, is also listed
C. ofŽ cinalis (Pietta et al. 1992). Fractionation by column chro- in the CLS. Calendula Powder, which conforms to the speciŽ -
matography and thin layer chromatography have been used to cations of the JCIC, has precedent for use in all CLS categories
determine some terpene derivatives of C. ofŽ cinalis (Gracza without restriction except eyeliner, lip, and oral preparations, for
1987). which there is no precedent for use.
 CALENDULA OFFICINALIS EXTRACT AND CALENDULA OFFICINALIS 15

TABLE 1
Calendula Extracts product formulation data (FDA 1998)

Total no. of formulations Total no. containing

Product category in category ingredient
Baby lotions, oils, powders, and creams 53 3
Other baby products 29 1
Bath oils, tablets, and salts 124 2
Bubble baths 200 1
Other bath preparations 159 1
Eyeliner 514 1
Eye shadow 506 1
Eye lotion 18 2
Other eye makeup preparations 120 5
Other fragrance preparations 148 2
Hair conditioners 636 9
Hair sprays (aerosol Ž xatives) 261 1
Hair straighteners 63 1
Permanent waves 192 4
Rinses (noncoloring) 40 3
Shampoos (noncoloring) 860 15
Tonics, dressings, and other hair-grooming aids 549 9
Other hair preparations 276 6
Blushers (all types) 238 2
Face powders 250 3
Foundations 287 1
Lipstick 790 3
Other makeup preparations 135 2
Bath soaps and detergents 385 9
Deodorants (underarm) 250 1
Other personal cleanliness products 291 2
Aftershave lotion 216 1
Shaving cream 139 1
Other shaving preparations 60 1
Cleansing preparations 653 14
Depilatories 28 1
Face and neck preparations (excluding shaving preparations) 263 6
Body and hand preparations (excluding shaving preparations) 796 11
Moisturizing preparations 769 12
Night preparations 188 7
Paste masks (mud packs) 255 7
Skin fresheners 184 6
Other skin care preparations 692 19
Suntan gels, creams, and liquids 136 2
1998 Total uses of Calendula Extracts 178

Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis do Noncosmetic

not appear in Annex II (list of substances which must not form C. ofŽ cinalis L. is generally recognized as safe (GRAS) in
part of the composition of cosmetic products), III (list of sub- spices and other natural seasonings and  avorings (FDA 1997).
stances which cosmetic products must not contain except sub- C. ofŽ cinalis has been used in traditional herbal medicine
ject to the restrictions and conditions laid down), or IV (list of (Gracza 1987), often because of its (reported) anti-in ammatory
coloring agents allowed for use in cosmetic products) of the activity (Boucaud-Maitre, Algernon, and Raynaud 1988; Della
Cosmetics Directive of the European Union (1995). Loggia et al. 1994; Bezákova et al. 1996).
16 COSMETIC INGREDIENT REVIEW

TABLE 2
Concentration of use data (FDA 1984)

Product category 5% – 10% 1% – 5% 0.1% – 1% Unknown Total

Calendula OfŽ cinalis Extract
Wave sets 1 1
Face powders 1 1
Foundations 1 1
Shaving cream (aerosol/brushless/lather ) 1 1
Face/body/hand preparations (excluding shaving) 1 5 7 13
Night preparations 2 2
Paste masks (mud packs) 1 1
Other skin care preparations 1 2 3
Suntan gels/creams/liquids 1 1
1984 Totals 1 3 8 12 24
Calendula OfŽ cinalis
Lipstick 1 1
Moisturizing products 1 1
Paste masks (mud packs) 1 1
1984 Totals 0 2 1 0 3

GENERAL BIOLOGY wounds of control animals. Also, the amount of “mature col-
lagen” Ž bers compared to “young collagen” Ž bers was much
Absorption, Distribution, Metabolism, Excretion
greater in the test than in the control animals.
Published data on the absorption, distribution, metabolism,
and excretion of Calendula OfŽ cinalis Extract and Calendula Immunologic Effects
OfŽ cinalis were not found.
Wagner et al. (1985) reported the isolation of polysaccha-
rides with molecular weights of 25,000 to >500,000 Da from
Dermal Effects an aqueous. NaOH extract of Calendula ofŽ cinalis L. that had
Standard skin wounds were induced on a depilated area of signiŽ cant immunostimulating activities according to the gran-
the backs of groups of 24 male Wistar rats, and the wounds were ulocytes and carbon clearance tests.
covered with 5% unguentum (control group), allantoin alone
(control group), or unguentum with C1 and C5 fractions isolated Cytotoxicity
from C. ofŽ cinalis  owers and allantoin (test group) to determine The cytotoxicity of Ž ve extracts of C. ofŽ cinalis that had
the effects on epithelialization (KloucÏ ek-Papova et al. 1982). The different compositions was evaluated using MRC5, Hep2, and
test and control materials were applied daily. Epithelialization Ehrlich cell lines (Boucaud-Maitre, Algernon, and Raynaud
was determined on days 0, 1, 3, 5, 14, and 21, wound exudate 1988). All the extracts were cytotoxic, but the activity using
was examined microscopically 8, 24, and 48 hours after wound 0.2 to 0.02 g/l of the extracts varied from 30% to 99% killed
in iction, and tissue samples from the wound were examined cells using the MRC5 cell line, from 2% to 99% killed cells us-
microscopically on day 10. There was “signiŽ cantly more inten- ing the Hep2 cell line, and from 10% to 100% cells killed using
sive epithelialization of the wounds” for animals of the control the Ehrlich cell line. Using saline solution as a control, the per-
group as compared to animals of the test groups, especially by centage of killed cells varied from 16% to 20%, 2% to 4%, and
day 14. Microscopic examination of the exudate from test ani- 0% to 2% for the MRC5, Hep2, and Ehrlich cell lines, respec-
mals detected a greater glycogen content in the “polynuclears,” tively. Antitumoral activity was also studied using the mouse
a “considerable number” of blast cells at 24 hours, the presence Ehrlich carcinoma. One of the extracts was inactive and three
of “numerous differentiated macrophages having a cytoplasm were “poorly active,” whereas “an absence of development of
rich in various inclusions” after 48 hours, and that “lymphoid the ascites was observed” with the most saponin-rich extract.
elements produce a more intensively nuclear (green)  uores-
cence” as compared with controls. At microscopic examination ANIMAL TOXICOLOGY
of the wound at day 10, a thinner leucocytic-necrotic torus was
observed on the surface of the wound of test animals as com- Acute Toxicity
pared to controls, and that “deŽ nite zones” of the wounds of Oral
test animals were almost entirely Ž lled with granulation tissue, The LD50 of Calendula OfŽ cinalis Extract for rats was
as opposed to insular development of granulation tissue in the >4640 mg/kg (CTFA 1980).
 CALENDULA OFFICINALIS EXTRACT AND CALENDULA OFFICINALIS 17

The LD50 , for rats, of a mixture containing Calendula OfŽ ci- 1984). During induction, intradermal injections of 0.05 ml of
nalis Extract (1% – 5%), soybean (Glycine Soja) oil (>50%), and 50% aqueous Freund’s complete adjuvant (FCA), 5% Calen-
tocopherol (<0.1%) was >20 ml/kg (Chemisches Laboratorium dula OfŽ cinalis Extract in propylene glycol, and 5% Calendula
Dr. Kurt Richter GmbH 1996). OfŽ cinalis Extract in 50% aqueous FCA were made to sites on
the upper back of each animal. A control group of 10 animals re-
Parenteral ceived the injections without the test material. During the booster
Using two to three albino mice per group, the intraperitoneal phase 1 week after induction, 20% Calendula OfŽ cinalis Extract
(IP) LD50 of Calendula OfŽ cinalis Extract was determined to be (a dose determined to be slightly irritating) was applied to the
300 mg/kg (Dhar et al. 1968). induction site for 48 hours under an occlusive patch. Two weeks
after application of the booster, the animals were challenged
with 5% and 10% Calendula OfŽ cinalis Extract that was ap-
Short-Term Toxicity
plied for 24 hours under an occlusive patch. The challenge sites
Groups of dd-mice were given 5, 10, or 20 ml/kg of a mix- were graded 24 and 48 hours after patch removal. Calendula
ture of Calendula OfŽ cinalis Extract, butylene glycol, and water OfŽ cinalis Extract, 5% or 10%, did not produce a sensitization
(percentages not speciŽ ed) for 14 days (Ichimaru Pharcos Co., reaction.
Ltd. 1994). The LD50 was >20 ml/kg. A mixture of Calendula OfŽ cinalis Extract (1% – 5%), soy-
bean (Glycine Soja) oil (>50%), and tocopherol (<0.1%) was
Subchronic Toxicity not sensitizing to guinea pigs (Chemisches Laboratorium Dr.
Published data on the subchronic toxicity of Calendula Of- Kurt Richter GmbH 1996).
Ž cinalis Extract and Calendula OfŽ cinalis were not found. The sensitization potential of a 50% aqueous solution of a
mixture containing Calendula OfŽ cinalis Extract, butylene gly-
col, and water (percentages not speciŽ ed) was determined in a
Chronic Toxicity maximization test using guinea pigs (Ichimaru Pharcos Co., Ltd.
Published data on the chronic toxicity of Calendula OfŽ ci- 1994). Erythema and edema were not observed.
nalis Extract and Calendula OfŽ cinalis were not found.
Photosensitization
Dermal Irritation
The phototoxicity potential of a 50% aqueous solution of
The dermal irritation potential of 10% aqueous Calendula a mixture containing Calendula OfŽ cinalis Extract, butylene
OfŽ cinalis Extract was determined in a single-insult occlusive glycol, and water (percentages not speciŽ ed) was determined
patch test (SIOPT) using nine rabbits (CTFA 1983). Calendula using six guinea pigs (Ichimaru Pharcos Co., Ltd. 1994). One-
OfŽ cinalis Extract, 10%, had a primary irritation index (PII) of tenth milliliter of the test article was applied and exposed to a
0.0 and was not an irritant. 15-minute minimal erythema dose. The mixture was not
An SIOPT was performed to determine the dermal irrita- phototoxic.
tion potential of an eye cream containing 1% Calendula OfŽ ci-
nalis Extract (CTFA 1986a). The eye cream produced minimal
Ocular Irritation
irritation.
The ocular irritation potential of 10% aqueous Calendula Of-
A mixture of Calendula OfŽ cinalis Extract (1% – 5%), soy-
Ž cinalis Extract was determined in a study using six rabbits
bean (Glycine Soja) oil (>50%), and tocopherol (<0.1%), tested
(CTFA 1983). The test article was instilled into the conjunctival
at 10% in liquid parafŽ n, was nonirritating to rabbits in a Draize
sac of the eye of each animal, and the eyes were not rinsed. Cal-
test (Chemisches Laboratorium Dr. Kurt Richter GmbH 1996).
endula OfŽ cinalis Extract, 10%, produced minimal irritation.
The irritation potential of a mixture containing Calendula
The ocular irritation potential of eye creams containing 1%
OfŽ cinalis Extract, butylene glycol, and water (percentages not
Calendula OfŽ cinalis Extract was also determined (CTFA
speciŽ ed) was determined in a Draize test in which 0.5 ml of
1986a). The eye creams produced no to minimal irritation.
the mixture was applied to intact and abraded skin of six albino
A mixture of Calendula OfŽ cinalis Extract (1% – 5%), soy-
rabbits (Ichimaru Pharcos Co., Ltd. 1994). The test sites were
bean (Glycine Soja) oil (>50%), and tocopherol (<0.1%), tested
scored 4, 24, and 48 hours after application. Very slight erythema
at 10% in liquid parafŽ n, was nonirritating to rabbit eyes in a
was observed after 4 hours. The mixture, 0.5 ml, was also applied
Draize test (Chemisches Laboratorium Dr. Kurt Richter GmbH
19 times to the skin of Ž ve guinea pigs over a 4-week period.
1996).
Erythema and edema were not observed.
The ocular irritation potential of a mixture containing Calen-
dula OfŽ cinalis Extract, butylene glycol, and water (percentages
Sensitization not speciŽ ed) was determined in a Draize test in which 0.1 ml
The sensitization potential of Calendula OfŽ cinalis Extract of the mixture was applied to the conjunctival sacs of six al-
was determined in a modiŽ ed Magnusson-Kligman maximiza- bino rabbits (Ichimaru Pharcos Co., Ltd. 1994). A conjunctival
tion test using 10 female Dunkin Hartley guinea pigs (CTFA reaction was observed in one rabbit.
18 COSMETIC INGREDIENT REVIEW

REPRODUCTIVE AND DEVELOPMENTAL TOXICITY 24 and 48 hours after patch removal. Erythematous reactions
Published data on the reproductive and developmental toxi- were not observed during induction or at challenge, and an eye
city of Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis cream containing 1% Calendula OfŽ cinalis Extract was not a
were not found. sensitizer.
An RIPT of a cosmetic formulation containing 1% Calen-
dula OfŽ cinalis Extract, using the same procedures described
GENOTOXICITY above, was completed using 102 subjects (TKL Research 1987).
An Ames test using Salmonella typhimurium TA98 was per- One subject had reactions at challenge indicative of a possible
formed without and with metabolic activation to determine the sensitization response. Upon rechallenge using occlusive and
mutagenic potential of six saponins isolated from the dried  ow- semiocclusive patches, the reactions were considered indicative
ers of C. ofŽ cinalis L. (Elias et al. 1990). The saponins, 80 to of irritation and not sensitization. A cosmetic formulation con-
200 ¹g/tube, were not mutagenic. The saponins, ·400 ¹g, also taining 1% Calendula OfŽ cinalis Extract was not a sensitizer.
were not toxic to S. typhimurium TA97, TA98, TA100, or TA102,
without or with metabolic activation in a preliminary spot test.
Predictive Testing
A somatic mutation and recombination test using Drosophila
melanogaster was performed to determine the genotoxic poten- A multicenter sensitization study using 119 subjects with
tial of 20% and 40% C. ofŽ cinalis herbal tea extract (Graf et al. contact allergic dermatitis was performed according to inter-
1994). The C. ofŽ cinalis tea was not genotoxic. Two  avonols, nationally accepted methods using the European standard series
quercetin and rutin, had weak genotoxic activity. and a number of cosmetic ingredients, including 10% Calendula
extract in alcohol (de Groot et al. 1988). The test materials were
applied for 2 days using van der Bend patch test chambers; the
CARCINOGENICITY test sites were scored 20 minutes and 1 and 2 days after removal.
Published data on the carcinogenic potential of Calendula Calendula extract caused a positive reaction in one subject.
OfŽ cinalis Extract and Calendula OfŽ cinalis were not found. A series of ointments, one of which contained calendula tinc-
ture, 10%, the European standard series, and the components of
the ointment bases, that is, petrolatum, liquid parafŽ n, wool fat,
CLINICAL ASSESSMENT OF SAFETY and chlorophyll, were evaluated for their sensitization potential
Dermal Irritation using 1032 subjects from six patch test clinics (Bruynzeel et al.
1992). Two subjects had positive reactions to the calendula tinc-
The irritation potential of a cosmetic formulation containing
ture; one of these subjects also had a positive reaction to wool
1% Calendula OfŽ cinalis Extract was performed using 14 sub-
fat. The researchers stated that “the relevance of the patch test re-
jects (CTFA 1986a). The cosmetic formulation had an irritation
actions is difŽ cult to evaluate” because the subjects often do not
index (PII) of 0.
know whether they have previously used the ointments. Also,
The cumulative irritation potential of a cosmetic formulation
“the number of reactions may be underestimated” because the
containing 1% Calendula OfŽ cinalis Extract was determined in
ointment base may not be a suitable vehicle for testing.
a study completed using 13 subjects, 3 males and 10 females
(Hilltop Research 1986). The test material, 0.2 ml, was applied A patch test was performed using 15 subjects according to
the methods of the International Contact Dermatitis Research
to a site on the back of each subject for 23 hours under an occlu-
Group (ICDRG) with the European standard series and some
sive patch daily for 20 days. The test sites were scored 1 hour
Compositae allergens, including 10% calendula in petrolatum
after patch removal. A cosmetic formulation containing 1% Cal-
(Wrangsjö, Ros, and Wahlberg 1990). The Compositae allergens
endula OfŽ cinalis Extract was considered a “mild material.”
were applied for 24 hours, and the test sites were scored after
In a 4-day minicumulative irritation test, a cosmetic eye prod-
20 and 60 minutes and 48 and 96 hours. Calendula, as both the
uct containing 1% Calendula OfŽ cinalis Extract applied under
plant extract and with pollen “as is,” produced a positive result
an occlusive patch had a PII of 0.24 (CTFA 1990). (Additional
in one of 15 subjects. The  ower of Calendula was tested “as
details not given.)
is,” either fresh or deep frozen for 6 months, and it produced a
positive reaction in the one patient tested.
Sensitization Commercial-grade absolute of calendula, 1% in petrolatum,
The sensitization potential of an eye cream containing 1% was applied to three subjects that were “contact-sensitive” to
Calendula OfŽ cinalis Extract was determined in a repeated- numerous Compositae species and sesquiterpene lactones and
insult patch test (RIPT) completed using 109 subjects, 11 males to six eczema patients (Rodr´õ quez and Mitchell 1977). (An ab-
and 98 females (CTFA 1986b). The eye cream, 0.1 ml, was ap- solute is a highly concentrated reŽ ned perfume material, usually
plied under an occlusive patch for 24 hours to a test site on the liquid, that has undergone at least two extractions; it is obtained
back of each subject 3 days per week for 3 weeks. Following a by alcohol extraction from concretes. A concrete is a solid, waxy
2-week nontreatment period, a challenge patch was applied to material extracted from non- or low-resinous material; natural
a previously unpatched site for 24 hours. The site was scored raw materials for concretes are usually prepared from vegetative
 CALENDULA OFFICINALIS EXTRACT AND CALENDULA OFFICINALIS 19

materials extracted from previously live tissue.) Positive reac- tive human patch testing, positive reactions to Calendula were
tions were not observed. seen in a small number of patients.
A sesquiterpene lactone mix, 0.1% petrolatum, was included
in a standard patch test series and 686 patients were patch-tested DISCUSSION
with the series (Paulsen, Andersen, and Hausen 1993). Seventy- Section 1, paragraph (p), of the Cosmetic Ingredient Review
nine patients who had positive reactions to the mix or who were (CIR) Procedures states that “A lack of information about an in-
suspected of having a Compositae dermatitis were tested with a gredient shall not be enough to justify a determination of safety.”
Compositae mix, 6% petrolatum. The test materials were applied In accordance with Section 30( j)(2)(A) of the Procedures, the
under occlusive patches to the backs of the patients, and the sites Expert Panel informed the public of its decision that the data
were scored on days 2, 3, or 4, and sometimes on days 5 to 7, on Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis were
according to the methods of the ICDRG. Thirty-one patients insufŽ cient to determine whether Calendula OfŽ cinalis Extract
had positive reactions to one or both mixes. One patient with and Calendula OfŽ cinalis were either safe or unsafe. The Ex-
Compositae allergy was patch tested with 10.0% C. ofŽ cinalis pert Panel released a Notice of InsufŽ cient Data Announcement
L. had no response. on June 6, 1997, outlining the data needed to assess the safety
of Calendula OfŽ cinalis Extract and Calendula OfŽ cinalis. The
SUMMARY types of data still required for each ingredient include2
Calendula OfŽ cinalis Extract is an extract of the calendula, C. 1. Current concentration of use data.
ofŽ cinalis, and Calendula OfŽ cinalis is a plant material derived 2. Function in cosmetics.
from the  owers of the calendula. In 1998, it was reported to the 3. UV absorption data; if absorption occurs in the UVA or UVB
FDA that Calendula OfŽ cinalis Extract was used in 178 cosmetic range, photosensitization data are needed.
formulations; in 1984, it was used at concentrations of ·10% 4. Gross pathology and histopathology in skin and other
and at unknown concentrations. Data submitted by the cosmetics major organ systems associated with repeated dermal
industry reported that one company uses Calendula OfŽ cinalis exposures. 3
Extract at concentrations of <0.5% in a styling gel, a shampoo, 5. Dermal reproductive/developmental toxicity data.3
and a cream rinse, another company uses it at a concentration of 6. Inhalation toxicity data, especially addressing the concentra-
0.2 weight %, and a supplier reported that a mixture of Calendula tion, amount delivered, and particle size.
OfŽ cinalis Extract (10% to 25%) and propylene glycol (>75%) 7. Genotoxicity testing in a mammalian system; if positive, a
is used at 1% to 10% in cosmetic products. Calendula OfŽ cinalis 2-year dermal carcinogenicity assay performed using Na-
was not reported to be used. tional Toxicology Program (NTP) methods in needed.
The oral and IP LD50 values for rats and mice of Calendula
OfŽ cinalis Extract were >4640 and 300 mg/kg, respectively. The Expert Panel originally also requested information on the
The oral LD50 , for rats, of a mixture containing Calendula Of- presence of contaminants. Some data were received and sum-
Ž cinalis Extract, soybean (Glycine Soja) oil, and tocopherol and marized in the report. The Expert Panel expects that pesticide
for mice, of a mixture containing Calendula OfŽ cinalis Extract, residues would be kept to a minimum.
butylene glycol, and water was >20 ml/kg. In SIOPTs, Calen- No offer to supply the needed data was received. In ac-
dula OfŽ cinalis Extract, 10%, was nonirritating and an eye cream cordance with Section 45 of the CIR Procedures, the Expert
containing 1% Calendula OfŽ cinalis Extract was minimally ir- Panel has issued a Final Report—InsufŽ cient Data. When the
ritating; mixtures that contained Calendula OfŽ cinalis Extract requested data are available, the Expert Panel will reconsider the
were not irritants. Calendula OfŽ cinalis Extract and mixtures Final Report in accordance with Section 46 of the CIR Proce-
containing Calendula OfŽ cinalis Extract were not sensitizers in dures, Amendment of a Final Report.
tests using guinea pigs. A mixture containing Calendula OfŽ c-
inalis Extract, butylene glycol, and water was not phototoxic. CONCLUSION
Calendula OfŽ cinalis Extract, 10%, caused minimal ocular irri- The CIR Expert Panel concludes that the available data are
tation, eye creams containing 1% Calendula OfŽ cinalis Extract insufŽ cient to support the safety of Calendula OfŽ cinalis Extract
caused no to minimal irritation, and mixtures containing Calen- and Calendula OfŽ cinalis for use in cosmetic products.
dula OfŽ cinalis Extract produced no or little irritation.
Six saponins isolated from the dried  owers of C. ofŽ cinalis
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Clinical testing of cosmetic formulations containing 1% Cal-
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not very irritating. In RIPTs, cosmetic formulations containing 3 These are data that would be expected from what is commonly referred to
1% Calendula OfŽ cinalis Extract were not sensitizing. In predic- as a “28-day dermal toxicity study.”
20 COSMETIC INGREDIENT REVIEW

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4 Available for review: Director, Cosmetic Ingredient Review, 1101 17th

Street, NW, Suite 310, Washington, DC 20036, USA.