Registration Dossier

Administrative data

Description of key information

Oral toxicity:

- 90-day rat (OECD 408). NOAEL: 450 mg/kg bw/day

Dermal toxicity:

- 28-day rat (OECD 410). NOAEL: 5000 mg/kg bw/day

- 21-day rabbit (EPA OPPTS 870.3200). NOAEL: 1500 mg/kg bw/day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, but study scope does not cover a full OECD 407 study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Uvinul MC 80 N was administered in the diet to groups of 13 female Wistar rats at nominal doses of 0 and 1000 mg/kg bw/day. The test substance concentration in the diet was adjusted weekly in order to achieve the intended dose level. Food consumption and body weights were determined weekly. The animals were examined for signs of toxicity or mortality at least once a day. Vaginal smears for estrus cycle determination were prepared and evaluated each day. From day 28 onwards blood was taken when the animals were in the proestrus cycle and the following hormones were determined: T3, T4, thyroid-stimulating hormone, prolactin, follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone. All animals were assessed by gross pathology. No histopathological examinations were performed.
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 11 to 13 weeks
- Weight at study initiation: mean weights per group 195.8 g and 196.8 g
- Housing: Singly in type DK III stainless steel wire mesh cages supplied by Becker & Co., Castrop-Rauxel, Germany (floor area about 800 cm²). Underneath the cages, disposable waste trays were used. type 3/4 dustfree embedding, supplied by SSNIFF was used.
- Diet: basic maintenance diet for mice and rats, meal "GLP", ad libitum
- Water: ad libitum
- Acclimation period: 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 hours light from 06.00 a.m. - 06.00 p.m., 12 hours dark from 06.00 p.m. - 06.00 a.m.

IN-LIFE DATES: From: 23.04.2003 To: 28.05.2003
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): basic maintenance diet
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
35 days
Frequency of treatment:
daily
Dose / conc.:
0 mg/kg diet
Remarks:
Basis: nominal in diet
Dose / conc.:
1 000 mg/kg diet
Remarks:
Basis: nominal in diet
No. of animals per sex per dose:
13
Control animals:
yes, plain diet
Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice a day (working days), once a day (week-ends, holidays)

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
Substance intake for day x= (FCx * C)/BWx
BWx= body weight on day x [g]
FCx = mean daily food consumption on day x [g]
C = concentration in the diet on day x [mg/kg]

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: Yes
- Animals fated: no
- How many animals: 10 animals
- Parameters checked: Hormones
* total triiodothyronine (T3)
* total thyroxine (T4)
* thyroid-stimulating hormone (TSH)
* prolactin (PRL)
* follicle-stimulating hormone (FSH)
* luteinizing hormone (LH)
* estradiol (E2)
* progesterone (PROG)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: No
Organ weights (ovaries)
Other examinations:
Estrus cycle determination
Statistics:
Body weight, food consumption, estrus stages: DUNNETT's test (two-sided) vs control group
Clinical patholgy: Wilcoxon-test (two- sided) vs control group
Organ weight: Wilcoxon-test vs control group
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weight was statistically significantly decreased from day 21 to day 35. The value on day 35 was 4.3 % below control.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In test group, food consumption was statistically significantly reduced on day 7 (5.5 % below control).
Food efficiency:
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Slight but statistically significantly increased thyroxine (T4) concentrations (+ 19 %) were found in the serum of the treated female rats. No effects were observed in the other thyroid hormones of the treated animals. Moreover, the test compound did not affect prolactin, follicle-stimulating hormone, luteinizing hormone, estradiol and progesterone level in the serum of the treated rats.
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
The only gross lesion noted was a cyst in the ovaries of one animal in the treated group.
This finding was regarded as incidental and unrelated to treatment.
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Key result
Dose descriptor:
LOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Key result
Critical effects observed:
not specified
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 March 1983 - 15-17 June 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted according to OECD guideline 408 and under GLP conditions. Report is well-documented.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
number of animals in high-dose group (n=12) due to recovery experiment (n=6)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Füllinsdorf Albino SPF (outbred)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Institute of biological and medical research, Füllinsdorf, Switzerland
- Age at study initiation: about 7 weeks
- Weight at study initiation:
Males: about 162 gr.
Females: about 136 gr.
- Housing: 2 rats per stainless-steel wire-mesh cage
- Diet (e.g. ad libitum): Ad libitum, powdered complete rodent breeding diet
- Water (e.g. ad libitum): Ad libitum, tap water
- Acclimation period: minimum 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 55 +/- 10
- Air changes (per hr): 20-25
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): powdered complete rodent breeding diet (NAFAG 850).
The feed mixture was prepared weekly by mixing the test compound with powdered diet for 10-15 min. in a mixing machine.

VEHICLE
- No vehicle used (unchanged diet)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of active ingredient found after preparation of the mixture corresponds well to the added concentration (88-110% of nominal).
The standard deviation of the feed mixtures is at less than 6.3%.
Duration of treatment / exposure:
minimum 90 days
Frequency of treatment:
continuous
Remarks:
Doses / Concentrations:
200, 450, 1000 mg/kg/day
Basis:
nominal in diet
No. of animals per sex per dose:
12
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Based on the results of a preliminary dose-range finding study, which covered the dose-levels of 175, 350 and 700 mg/kg/day, the dosing schedule for the 13-week study was chosen.
- Rationale for animal assignment (if not random): At random
- Rationale for selecting satellite groups: None selected
Positive control:
None
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked:
Morbidity/mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly for each group and sex.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes, calculated from the average feed intake.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: At weeks 7 and 12
- Dose groups that were examined: Control and high-dose animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At weeks 2, 6 and 14
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 6/sex/dose
- Parameters checked:
Red Blood Cells (RBC)
Haemoglobin (HB)
Mean Corpuscular Volume (MCV)
Packed Cel]. Volume (PCV)
Mean Corpuscular Haemoglobin (MCH)
Mean Corpuscular Haemoglobin Concentration (MCHC)
Reticulocytes (RETI) Method of Dacie
White Blood Cells (WBC)
Differential Cell Count:
- Polymorphonuclear Neutrophil (SEG)
- Juvenile Neutrophil (BAND)
- Lymphocyte (LYMPH)
- Monocyte (MONO)
- Eosinophil (EOS)
- Basophil (BASO)
- Immature White Blood Cell (IMMAT WBC)
- Immature Lymphocyte (IMMAT LYMPH)
- Plasma Cell (PLASM CELL)
- Nucleated Red Blood Cell (NRBC)
Thrombocytes (THROM)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At weeks -1, 6, 14 and 19
- Animals fasted: No data
- How many animals: No data
- Parameters checked:
Total bilirubin
Urea
Glucose
Cholesterol
Serum-Sodium
Serum-Potassium
Serum-Total-Proteins
Serum-Electrophoresis
Alkaline phosphatase
Aspartate aminotransferase
Alanine aminotransferase
Glutamate dehydrogenase

URINALYSIS: Yes
- Time schedule for collection of urine: At weeks 4 and 12
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, during collection
- Parameters checked:
Color
Volume in ml
Specific gravity
pH
Protein content
Glucose content
Occult blood
Bilirubin, Urobilinogen
Ketone Bodies
Microscopic composition
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were subjected to full gross necropsy

ORGAN WEIGHTS:
Brain, heart, liver, kidneys, testes, ovaries, adrenal glands were weight wet (paired organs = weight of both organs).

HISTOPATHOLOGY: Yes
All gross lesions
Pituitary
Thyroids
Salivary glands
Thymus
Adrenal glands
Lungs with bronchi
Trachea
Heart
Aorta
Liver
Esophagus
Stomach
Duodenum
Je junum
Ileum
Caecum
Colon
Rectum
Pancreas
Mesenteric lymph node
Spleen
Kidneys
Urinary bladder
Testes
Epididymis
Prostate
Seminal vesicle
Ovaries
Uterus
Brain
Nervus ischiadicus
Eyes
Bone
Bone marrow
Muscle
Skin
Mammary glands
Other examinations:
Not relevant
Statistics:
Results of hematology and blood chemistry: Dunn-test
Body and organ weights: Growth rate, adjusted organweights and Jonckheere- and U-test.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Only observation in connection with the treatment was the occurrence of soiled tails especially in high-dose groups.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No mortalities related to the treatment with the test compound occurred during the study.
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
None of the effects found were significantly or treatment related. For some observed effects individual values remained within the normal ranges for rats.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There was a slight increase of the absolute and the allometrically adjusted weights of kidneys and livers of treated males, but only the weights of the kidneys at the high-dose were statistically significant different from those of the controls. After recovery no weight difference was found, which indicates most probably that the weight difference reflects the adaptation to the eliminatory task. For females, no effects were found.
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Microscopical changes related to the treatment were confined to the livers. They consisted of the reduction of the glycogen content, especially in high-dose animals, which was accompanied by the shrinkage of hepatocytes in some males and females and a slight non-significant increase of the phagocytized iron positive material in the Kupffer cells of mid and high-dose females. After the recovery period, there was no obvious difference between the livers of high-dose and control animals.
Histopathological findings: neoplastic:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
450 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
clinical signs
organ weights and organ / body weight ratios
histopathology: non-neoplastic
Dose descriptor:
LOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: overall effects
Key result
Critical effects observed:
not specified
Conclusions:
Ethylhexyl methoxycinnamate was well tolerated at any dose-level under the conditions of this 13-week oral feeding study in rats. A NOAEL of 450 mg/kg/day was established. Based on these results, ethylhexyl methoxycinnamate does not need to be classified as toxic after repeated exposure based on the criteria outlined in Annex I of 1272/2008/EC.
Executive summary:

Ethylhexyl Methoxycinnamate, an UV-B absorber used in sunscreens was orally administered to rats (12/sex/group) at the doses 0 (control), 200, 450 and 1000 mg/kg/day, 7 days per week for at least 13 weeks. At the termination of the treatment, half of the controls and of the high-dose rats were allowed 5 additionally weeks on normal diet prior to sacrifice.

Body weight, feed consumption and signs of toxicity were recorded weekly. Ophthalmoscopy and urine analysis were performed twice during the study. Blood chemical and hematological investigations were carried out at the beginning, during and at the end of the treatment period. An additional blood chemical investigation was performed after the recovery period. Post mortem investigations comprised full autopsy, organ weight determinations and histological examination.

The feed intake and the body weight development of treated animals were similar to those of controls. No symptoms indicative of pathologic conditions, ophthalmological abnormalities or mortalities as consequence of the treatment with the test compound were recorded during the study. Laboratory investigations in high-dose females (1000 mg/kg/day) revealed an increase of the plasma activity of GLDH which was reversed after the recovery period. The absolute as well as the allometrically adjusted weights of the kidneys were slightly increased in males of the high-dose group. No deviations of the weights were found after the recovery period, thus indicating an adaptive change. The glycogen content of the livers in high-dose animals was reduced and in 5 of 12 animals it was accompanied by slight shrinkage of the hepatocytes. In females of mid and high-dose group the amount of the iron positive material phagocytized by Kupffer cells was slightly increased. These conditions were reversed after the recovery period.

There was no obvious effect related to the treatment, which was detectable by the hematological, blood chemical and urine parameters at the mid- (450 mg/kg/day) and low-dose (200 mg/kg/day) levels. A slight increase of the iron positive material phagocytized by the Kupffer cells was observed in mid-dose females.

It is concluded that the treatment with Ethylhexyl Methoxycinnamate was well tolerated at any dose level and that under the conditions of this study only minor and reversible changes occur at the doselevel of 1000 mg/kg/day, whereas the dose of 450 mg/kg/day does not induce any adverse effect in the rat. Based on these results, ethylhexyl methoxycinnamate does not need to be classified as toxic after repeated exposure based on the criteria outlined in regulation 1272/2008/EC.

Endpoint conclusion
Dose descriptor:
NOAEL
450 mg/kg bw/day

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 1979 - August 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was not conducted under GLP and according to an OECD guideline. However, the experimental design resembles OECD guideline 410 and the report is well documented.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Deviations:
not applicable
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Bantin & Kingman Ltd., Grimston, Aldbrough, Hull
- Weight at study initiation:
Male: 181 - 232 g
Female: 158 - 196 g
- Housing: Individually, in solid bottomed plastic cages
- Diet (e.g. ad libitum): Ad libitum, pelleted
- Water (e.g. ad libitum): Ad libitum, water bottles
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 25
- Humidity (%): 40 - 60
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on exposure:
TEST SITE
- Area of exposure: Back and flanks
- Type of wrap if used: Aluminium foil with a strip of impermeable plaster
- Time intervals for shavings or clipplings: 24 hours prior to treatment

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
Group 1: 0 mL/kg bw/day
Group 2: 0.5 mL/kg bw/day (= 500 mg/kg/day)
Group 3: 1.5 mL/kg bw/day (= 1500 mg/kg/day)
Group 4: 5.0 mL/kg bw/day (= 5000 mg/kg/day)
- Concentration (if solution): 1000 mg/mL (specific gravity: 1.011)
- Constant concentration used: Yes
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not relevant
Duration of treatment / exposure:
28 days, 6 hours contact period/day
Frequency of treatment:
Once daily
Dose / conc.:
500 mg/kg bw/day (nominal)
Dose / conc.:
1 500 mg/kg bw/day (nominal)
Dose / conc.:
5 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Intact skin: 5
Abraded skin: 5
Control animals:
yes
Details on study design:
- Dose selection rationale: selected after examination of data from a 7 day dose range-finding study
- Section schedule rationale (if not random): Animals were allocated randomly throughout the whole study
Positive control:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations:
Signs of toxicity
Ill health
Behavioural change

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Daily
Erythema and oedema were scored on a scale from 0 - 4.
All-or-nothing scale was used for skin reactions concerning desquamation, fissuring and atonia.

BODY WEIGHT: Yes
- Time schedule for examinations: prior to treatment on first day and thereafter on every Monday and Thursday

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Pre-dose and pre-termination during week 4
- Dose groups that were examined: All animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Prior to commencement of therapy and during week 4 of study
- Anaesthetic used for blood collection: Yes (diethyl ether)
- Animals fasted: Yes, approx. 16 hours
- How many animals: All animals
- Parameters checked:
Haemoglobin (Hb)
Packed cell volume (PCV)
Red blood cell count (RBC)
Mean cell haemoglobin (MCH)
Mean cell volume (MCV)
Mean cell haemoglobin concentration (MCHC)
Total white blood cell count (WBC)
Differential white blood cell count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Prior to commencement of therapy and during week 4 of study
- Animals fasted: No data
- How many animals: All animals
- Parameters checked:
glucose
blood urea nitrogen (BUN)
glutamate-oxaloacetate transaminase activity (GOT)
glutamate-pyruvate transaminase activity (GPT)
alkaline phosphatase activity (Alk. P)
sodium ions
calcium ions
potassium ions
total protein
albumin
albumin/globulin ratio (A/G ratio)
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Organ weigths: adrenals, brain, heart, kidneys, liver, ovaries, pituitary, testes, thyroids (with parathyroids)

HISTOPATHOLOGY: Yes
Following tissues:
adrenals
aorta (arch and anterior abdominal)
brain
caecum
colon
duodenum
ep ididymus
eye and optic nerve
heart
ileum
Jejunum
kidneys
liver
lungs
lymph nodes (mandibular and mesenteric)
skeletal muscle (quadriceps)
all unusual lesions
oesophagus
ovaries
pancreas
pituitary
prostate
rib and bone marrow
sciatic nerve
skin and mammary gland
spinal cord (lumbar region)
spleen
stomach
salivary gland (submaxillary)
t est es
trachea
treated and untreated skin
thytnus
thyroids
tongue
urinary bladder
uterus (corpus and cervix)
adrenals
aorta (arch and anterior abdominal)
brain
caecum
colon
duodenum
ep ididymus
eye and optic nerve
heart
ileum
Jejunum
kidneys
liver
lungs
lymph nodes (mandibular and mesenteric)
skeletal muscle (quadriceps)
all unusual lesions
oesophagus
ovaries
pancreas
pituitary
prostate
rib and bone marrow
sciatic nerve
skin and mammary gland
spinal cord (lumbar region)
spleen
stomach
salivary gland (submaxillary)
testes
trachea
treated and untreated skin
thytnus
thyroids
tongue
urinary bladder
uterus (corpus and cervix)
Statistics:
Group mean values and standard deviations
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
Irritation of the skin at the application site was first noted prior to the third application. The severity of reaction was most marked in group 4 (highest dose) animals and in particluar the female animals. This erythema and oedema was accompanied by desquamation of the application site. In general the skin reactions noted in groups 2 and 3 appeared mild (lower dose groups) and little irritation was noted in control group 1.
The irritation reached a maximum at the end of the second week and then regressed. The skin appeared normal in all treated animals at termination, with the exception of one male animal in the highest dose group.
The reactions indicate that the test article is a low grade irritant under the experimental conditions.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
5 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: overall effects
Key result
Critical effects observed:
not specified
Conclusions:
The skin irritation reactions indicate that Ethylhexyl Methoxycinnamate is a low grade irritant under the experimental conditions. No evidence of any systemic toxicity by Ethylhexyl Methoxycinnamate was demonstrated. A NOAEL of 5000 mg/kg bw/day was established and the substance does not need to be classified as toxic after repeated exposure according to the criteria outlined in Annex I of 1272/2008/EC.
Executive summary:

Three groups of rats, each containing 5 male and 5 female animals with intact skin and 5 male and 5 female animals with abraded skin were treated by topical application with test article Ethylhexyl Methoxycinnamate at levels of 500, 1500 and 5000 mg/kg bw/day for 28 consecutive days. In addition a control group of 5 male and 5 female animals with intact skin and 5 male and 5 female animals with abraded skin was given a similar topical application of distilled water at a dose level of 5000 mg/kg bw/day for 28 days.

No animals died during the observation period in any group. No overt signs of toxicity were apparent in any treated group or control group during the observation period. No effect on body weight gain was apparent in any treated group compared with the control groups. The skin reactions observed suggest the test article is a low grade irritant under the experimental conditions used. No ocular defects related to treatment could be detected during an ophthalmoscopic examination at week 4. No treatment-induced effects were apparent in the haematology and blood chemistry parameters measured. No treatment related changes in individual organ weights or organ/body weight ratios were apparent. No evidence of systemic toxicity was noted at necropsy in any of the tissues and organs evaluated. Histological evaluation of the skin sites revealed a low grade epidermal proliferation in all groups.The degree of epidermal change tended to be dose-related, and males tended to show more epidermal change than females. There was no appreciable difference between intact and abraded skin sites, and no significant dermal inflammatory or fibrotic responses were seen in any rat.

Overall, a minimal to moderate epidermal proliferation in the absence of significant dermal changes suggested that the test article was a relatively low grade irritant under the prevailing experimental conditions. There was no evidence of systemic toxicity histologically in any of the other organs or tissues examined. A NOAEL of 5000 mg/kg bw/day was established and the substance does not need to be classified as toxic after repeated exposure according to the criteria outlined in Annex I of 1272/2008/EC.

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
June 1979 - July 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The EPA guideline for the registration of pesticides in the U.S. - the "sub-chronic 21 day dermal toxicity study" was used. Report is well-documented.
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.3200 (Repeated Dose Dermal Toxicity -21/28 Days)
Deviations:
yes
Remarks:
the used protocol was based on the EPA guideline
GLP compliance:
no
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Ranch Rabbits, Crawley Down, Sussex, England
- Weight at study initiation:
Males: 2.22 - 2.80 kg
Females: 2.23 - 2.95 kg
- Housing: Individually in grid floor cages
- Diet (e.g. ad libitum): Ad libitum, Ranch pellets
- Water (e.g. ad libitum): Ad libitum, filtered mains water
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 14 or above
- Air changes (per hr): Fan controlled air circulation
- Photoperiod (hrs dark / hrs light): 10/14
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on exposure:
TEST SITE
- Area of exposure: back and flanks (abraded or intact)
- % coverage: 10
- Type of wrap if used: gauze pad (10x10) on treated area using strip of impermeable adhesive plaster
- Time intervals for shavings or clipplings: every 48 hours

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Only wiping
- Time after start of exposure: 6 hours

TEST MATERIAL
- Amount(s) applied:
Control: 5.0 mL/kg/day (distilled water)
Test article groups: 0.5, 1.5 and 5.0 mL/kg/day (= 500, 1500 and 5000 mg/kg/day based on density Ethylhexyl Methoxycinnamate: 1.010 - 1.020)
- Concentration: Undiluted (1000 mg/mL)
- Constant volume or concentration used: no, adjusted weekly based on the most recent body weight
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not relevant
Duration of treatment / exposure:
21 days: 6 hours/day
Frequency of treatment:
Daily
Dose / conc.:
500 other: mL/kg bw
Dose / conc.:
1 500 other: mL/kg bw
Dose / conc.:
5 000 other: mL/kg bw
No. of animals per sex per dose:
10 (5 abraded and 5 intact skin)
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: The dose levels were selected after examination of the results obtained from a preliminary 7 day dose range finding study
- Rationale for animal assignment: At random
- Rationale for selecting satellite groups: No satellite groups
- Section schedule rationale: Animals were killed so that as far as possible equal numbers of male and female animals from each dose group were necropsied per day.
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations:
Signs of toxicity, behavioural change.

DERMAL IRRITATION: Yes
- Time schedule for examinations: Daily, prior to application of test article

BODY WEIGHT: Yes
- Time schedule for examinations: First day of study, followed by every Monday and Thursday during the rest of the study

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: predose and after 3 weeks
- How many animals: All
- Parameters checked:
haemoglobin (Eb)
packed cell volume (PCV)
red blood cell count (R.BC)
mean cell haemoglobin (MCH)
mean cell volume (MCV)
mean cell haemoglobin concentration (MCHC)
white blood cell count (WBC)
differential white blood cell count (neutrophils (N), lymphocytes (L), monocytes (M), eosinophils (E) and basophils (B))
platelet count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: predose and after 3 weeks
- How many animals: All
- Parameters checked:
glucose
cholesterol
blood urea nitrogen (BUN)
bilirubin
uric acid
ghitamate oxaloacetate transaininase (GOT)
glutamate pyruvate trarisaminase (GPT)
alkaline phosphatase (Alk.P)
lactate dehydrogenase (LDH)
sodium
calcium
potassium
chloride
total protein
albumin
A/G ratio

OTHER:
BONE MARROW ANALYSIS: Yes
Terminal bone marrow smears from the control group and high dose group were examined microscopically. A subjective assessment of the ratio of myeloid elements to erythoid elements was made.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Major tissues and organs were examined for presence of gross lesions

HISTOPATHOLOGY: Yes

Stained with haematoxylin and eosin for microscopic examination:
liver
kidneys
brain (3 levels)
pituitary
heart
thyroids
adrenals
testes
ovaries
treated and untreated skin as follows:
1. One section of untreated skin
2. One section of treated skin from the central area of application
3. One section of treated skin between the centre and periphery of the area of application
4. One sect ion of treated skin from the periphery of the area of application all unusual lesions

Additional sections of skin from the mammary region were examined in high dose group animals (group 4) to confirm microscopic findings in untreated skin sections.

Fixed and preserved:
adrenals
aorta (arch and anterior
brain
caecum
colon
duodenum
eye and optic nerve
gall bladder
heart
ileum
oesophagus
abdominal)
ovaries
pancreas
pituitary
prostate
rib and bone marrow
sciatic nerve
skin and mammary gland
spinal cord (lumbar region)

OTHER:
Organ weights of following organ were determined:
adrenals
brain
heart
kidneys
ovaries
pituitary
testes
thyroids
liver

Organ to body weight ratios were calculated.
Statistics:
Dose and sex group mean values and standard deviations were calculated
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Major clinical signs observed in treated animals during study were: lethargy, hunched posture, soiled and matted coats, alopecia, increased respiration, emaciation and conjunctival swelling. These were in general confined to the high dose group animals. No differences in the clinical symptoms were recorded between male and female animals, or between animals with intact and abraded skin within this group.
The lower dose groups showed soiling of fur, lethargy, a watery nasal discharge, periocular swelling and prominent ocular blood vessels.
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
Significant skin irritation was observed in all high, intermediate and low dose group animals during the study.
High dose group: slight to moderate erythema, slight oedema, slight to moderate desquamation, moderate cracking, slight atonia and folding of the skin. Similar for male/female and abraded/intact skin.
Intermediate and low dose group: very slight to slight erythema, very slight to slight oedema, slight desquamation, slight to moderate cracking and folding of the skin. Similar for male/female and abraded/intact skin.
Control group: very slight erythema only, mainly in abraded skin group.
Mortality:
mortality observed, treatment-related
Description (incidence):
Three high dose group animals died or were killed in extremis during experimental period. No further deaths occurred.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
High dose group: BW lower than those of intermediate, low and control group animals at all weightings (decrease at first application, weight loss not recovered thereafter. This was seen for male/female and abraded/intact skin.
Intermediate, low and control group: BW is generally similar (decrease at first application, healthy increase thereafter). This was seen for male/female and abraded/intact skin.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
High dose group: less than amount of food consumed by the other groups (especially during the first 2 weeks). This was seen for male/female and abraded/intact skin.
Intermediate, low and control groups: generally similar during study. This was seen for male/female and abraded/intact skin.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
High dose group: reversal of relative numbers of neutrophils (from low to high) and lymphocytes (from high to low) from predose to terminal period. Predose the counts were within the normal ranges.
Intermediate, low and control group: similar mean counts of neutrophils and lymphocytes from predose to terminal period. These are within the normal ranges.
There were no marked intergroup differences in the total white cell counts or in any of the other haematology paramters measured.
Mean platelet counts in predose and terminal samples from all groups were below the normal range expected. This is an unavoidable consequence of the drip bleeding method used to collect samples.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
High dose group: Plasma activity of alkaline phosphatase was depressed in blood samples taken terminally. Also a change in the level of plasma urea nitrogen was observed (elevated). Predose the concentration was within the distribution of levels of the lower dose groups.
Intermediate, low and control groups: Level of alkaline phosphatase within normal ranges. Mean levels for plasma urea nitrogen were similar in these groups.
No marked dose related changes occurred in any of the other clinical chemistry parameters measured.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
High dose group animals: Actual weights of liver, ovaries, tested and thyroids appeared depressed. Ratio with total weight indicated that this was linked to poor condition rather than to a toxic effect. Testicular retardation was however suggested to be treament-related.
Organ to body weight ratios also showed equivocal elevations of kidney and pituitary weight in high dose group animals.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Skin reaction in high, intermediate and low dose group animals involved both the epidermis and dermis and consisted essentially of slight to marked proliferation in hyperkeratosis, parakeratosis, hypergranulosis and acanthosis accompanied by generally low grade congestion, oedema and leucocyte infiltration in the superficial dermis. There seems to be a tendency towards a dose-related increase of effects. No difference was seen between male/female and abraded/intact skin.
No toxic effects in the intermediate and low dose groups were found.
Skin thickening in untreated samples was restricted mainly to the high dose group with a few minor cases in the intermediate dose group.
The most significant pathology findings in the high dose group animals were the generally poor bodily condition, indistinguishable thymus, a low incidence of grossly visible focal liver necrosis, depletion of glycogen vacuolation in the liver sections and more immature testes in these males than in the other groups.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
All treated groups: epidermal proliferative response accompanied with low grade inflammatory reaction on skin sites.
The degree of epidermal change tended to be dose-related, but there was no obvious difference between males and females or intact and abraded skin sites at any dose level.
A significant prominent epidermal reaction was seen in the high dose group animals.
Overall, the test article was found to be moderately irritating under the test conditions.
No evidence of systemic toxicity was found in intermediate or low dose group animals. High dose group animals were generally thin and in poor condition (3 animals died). Two losses were associated with respiratory conditions, one probably with enteric disturbances.
Histopathological findings: neoplastic:
not examined
Details on results:
OTHER FINDINGS
BONE MARROW
A subjective assessment of the ratio of myeloid elements to erythroid elements in terminal bone marrow smears taken from control and high dose group animals showed an apparent increase in the ratio in the high dose group. However, this observation was not supported by other haematology data which showed no evidence of a treatment related effect on the total red and white cell counts.
Key result
Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Key result
Critical effects observed:
not specified
Conclusions:
The grading of irritation reactions and microscopic observations during the study indicated ethylhexyl methoxycinnamate to be a moderate skin irritant under the prevailing conditions of the experiment. A NOAEL of 1500 mg/kg/day was established, indicating that ethylhexyl methoxycinnamate does not need to be classified as toxic after repeated exposure according to the criteria outlined in regulation 1272/2008/EC.
Executive summary:

Following a preliminary 7 day dose rangefinding study, 3 groups of 10 rabbits (5 male, 5 female) with intact skin and 3 groups of 10 rabbits (5 male, 5 female) with abraded skin were given occluded dermal doses of test article ethylhexyl methoxycinnamate at levels of 0.5, 1.5 and 5.0 mL/kg/day (= 500, 1500 and 5000 mg/kg bw/day) over a 21 day period. Two additional groups of 10 rabbits (5 male, 5 female) with intact and abraded skin were treated with distilled water at a volume of 5.0 mL/kg/day as controls.

Three high dose group animals (5000 mg/kg bw/day) died or were killed in extremis after 9 - 15 days of treatment. No further deaths occurred. Adverse clinical signs were mainly confined to animals in the high dose group and consisted of lethargy, hunched posture, soiled and matted fur, alopecia, increased respiration, emaciation and conjunctival swelling.

Skin irritation reactions resulting from treatment were erythema, oedema, desquamation, cracking, atonia and folding. The severity of these reactions varied between the groups, being in general, slight to moderate in high dose group animals (occasionally severe) and very slight to slight in intermediate and low dose group animals.

The body weights of animals in the high dose group were markedly lower than those of animals in the intermediate, low and control groups throughout the study.

Blood samples taken from high dose group animals after approximately 3 weeks of treatment showed an increase in the number of neutrophils, a corresponding decrease in the number of lymphocytes, depressed alkaline phosphatase activity and elevated levels of urea nitrogen.

Organ weights recorded at necropsy indicated a retardation of testicular growth in male high dose group animals.

Histological evaluation of the skin sites revealed an epidermal proliferative response in all treated groups accompanied by a low grade inflammatory reaction in the superficial dermis. The degree of epidermal change tended to be dose-related, but there was no obvious difference between males and females or between intact and abraded skin sites at any dose level. A significant feature of the high dose group was a prominent epidermal reaction in the samples of skin taken from untreated areas. Overall, the prominent epidermal proliferation plus the low grade dermal inflammatory reaction suggested that the test article was moderately irritant under the prevailing experimental conditions.

No evidence of systemic toxicity was found in intermediate or low dose group animals. High dose group animals were generally thin and in poor condition and 3 animals died or were killed because of illness before the end of the study. Two of these losses were associated with respiratory conditions and the third death probably resulted from enteric disturbances. Pathology findings in the surviving rabbits included diminished thymus, a low incidence of macroscopically observable focal liver necrosis, deleted liver glycogen and immature testes. These findings were considered to be related to the general debilitated condition of the rabbits rather than evidence of direct organotoxicity.

The grading of irritation reactions during the study indicated the test article to be a moderate skin irritant under the prevailing conditions of the experiment. This was supported by microscopic observations. Pathological findings in animals were contributed to ill-health (by skin irritation) rather than direct organotoxicity. This is also the case for the dose related changes in blood and liver parameters. A NOAEL of 1500 mg/kg/day was established, indicating that ethylhexyl methoxycinnamate does not need to be classified as toxic after repeated exposure according to the criteria outlined in regulation 1272/2008/EC.

Endpoint conclusion
Dose descriptor:
NOAEL
1 500 mg/kg bw/day

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose toxicity: oral

The 90-day oral toxicity study in rat (Camponovo, 1984, OECD 408) indicated that a dose of 1000 mg/kg/day induced liver changes (histopathology), resulting in a NOAEL of 450 mg/kg bw/day. This study was considered as key study.

A further supporting study undertaken was a hormone study in female Wistar rats (BASF SE, 2004) with administration at 1000 mg/kg bw/day in the diet over 4 weeks. Multiple hormone assays were undertaken. The rat is considered a very sensitive species for indirect thyroid hormone effects in comparison to man. Serum thyroid stimulating hormone (TSH) and total triiodothyronine (T3) were not affected. Slightly increased serum thyroxine (T4) concentrations were observed but, in the absence of effects on TSH or T3, ethylhexyl methoxycinnamate was not considered to be a true thyroid modulator or thyroid toxicant. In the other hormone investigations, in particular for estradiol and progesterone, no treatment-related effects were observed. (further studies on the endocrine system were performed: see section 7.8.3)

Repeated dose toxicity: dermal

A 28-day dermal toxicity study in rat (Keller, 1980, equivalent of OECD 410) indicated only dermal irritation, but no systemic toxicity. Therefore, a NOAEL of 5000 mg/kg/day was established. A 21-day dermal toxicity study in rabbit (Keller, 1980, non-OECD, but EPA OPPTS 870.3200) indicated multiple effects, which were attributed to ill-health (skin irritation) rather than direct organotoxicity. This resulted in a NOAEL of 1500 mg/kg/day. As the effects were not seen in other treatment groups or the control group, the effects could be caused by the high dose of ethylhexyl methoxycinnamate. The NOAEL of 1500 mg/kg bw/day is therefore chosen as the key parameter for repeated dermal exposure.


Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: liver; glandular: thyroids


Repeated dose toxicity: dermal - systemic effects (target organ) cardiovascular / hematological: bone marrow; cardiovascular / hematological: hematopoiesis; digestive: liver; other: skin; other: all gross lesions and masses

Justification for classification or non-classification

Based on the key parameters (NOAELs) given above, ethylhexyl methoxycinnamate does not need to be classified for repeated dose toxicity, as the NOAEL values are significantly higher than the guidance values for "Specific Target Organ Toxicity (STOT) - Repeated Exposure (RE) Category 2" as outlined in regulation 1272/2008/EC.

Guidance values used:

- Oral, rat, 90-days: 100 mg/kg bw/day

- Dermal, rat/rabbit, 28-days: 600 mg/kg bw/day (corrected by a factor of 3 for 90-days exposure, as indicated in regulation 1272/2008/EC)