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Toxicological information

Repeated dose toxicity: dermal

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Administrative data

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1981-02-03 to 1981-06-07
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
sub-chronic toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1981-02-03 to 1981-06-07
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose:
read-across source
Related information:
Composition 1
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
GLP compliance:
not specified
Test material information:
Composition 1
Specific details on test material used for the study:
5% Hexyl Cinnamic Aldehyde in phenyl ethyl alcohol
The specific gravity for all the compound was 1.00.
The test articles were stored for the duration of the study at ambient temperature and humidity.
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Ace Animals, Inc., P.O. Box 122, Boyertown, PA
- Females (if applicable) nulliparous and non-pregnant: Not specified
- Age at study initiation: Not specified
- Weight at study initiation: 75-100g
- Fasting period before study: Not specified
- Housing: The animals were housed/caged (Rats will be individually housed in wire mesh cages in a temperature controlled room reserved exclusively for this test. Cages will meet the minimum size requirement specified in DHEU Publication No. (NIH) 77-23, entitled "Guide for the Care and Use of Laboratory Animals) for 3 days after receipt to facilitate adjustment to the watering system. Thereafter, they were individually housed.
- Diet (e.g. ad libitum): Purina Rat Mash (#5001) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: The animals were kept in a room reserved for this test from date of receipt until dosing was initiated. During this period, the rats were observed daily. Body weights where recorded.

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The animal room was temperature and humidity controlled
- Humidity (%): The animal room was temperature and humidity controlled
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): Light was automatically controlled on a 12 hour light/dark cycle.

IN-LIFE DATES: From: 1981-02-03 To: 1981-06-07
Type of coverage:
not specified
Vehicle:
other: Phenyl ether alcohol
Details on exposure:
The test articles were applied topically, 7 days/week for 90 days by syringe and ball-tipped needle. The backs were divided into a seven (7) area grid and a separate area was treated daily to minimize local irritation. The rats were clipped once a week or as needed. Dosage volumes were based on weekly body weights.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
7 days/week for 90 days
Frequency of treatment:
90 days
Dose / conc.:
25 mg/kg bw/day (nominal)
No. of animals per sex per dose:
17/sex
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Not specified
- Rationale for animal assignment (if not random): Rats in good health were assigned to study groups by random numbers generated by a program in the TI 59 programmable calculator. Ear tags were secured after randomization.
Observations and examinations performed and frequency:
The rats were observed daily for signs of toxicity, pharmacological effects and mortality
Body Weights were recorded weekly
Dermal observations were recorded daily.
Clinical studies: The following parameters were measured at termination in all rats
hematology: erythrocytes, hemoglobin, hematocrit and total and differential leucocytes.
clinical chemistry: glucose, blood urea nitrogen, cholesterol, total bilirubin, uric acid, lactic dehydrogenase, albumin total protein. calcium, phosphorus. alkaline phosphatase, serum glutamic-oxaloacetic transaminase and serumglutamic - pyruvic transaminase.
urinalysis: appearance, color, specific gravity, pH, protein, glucose, ketone, bilirubin, occult blood and microscopic examination of the sediment

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily. The rats were observed daily for signs of toxicity, pharmacological effects and mortality.

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION: No data
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Not specified
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: erythrocytes, hemoglobin, hematocrit and total and differential leucocytes.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Not specified
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: glucose, blood urea nitrogen, cholesterol, total bilirubin, uric acid, lactic dehydrogenase, albumin, total protein. calcium, phosphorus. alkaline phosphatase, serum glutamic-oxaloacetic transaminase and serum glutamic-pyruvic transaminase

URINALYSIS: Yes
- Time schedule for collection of urine: Not specified
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters examined: appearance, color, specific gravity, pH, protein, glucose, ketone, bilirubin, occult blood and microscopic examination of the sediment

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

a. Animals were anesthetised with ether and exsanquinated on Day 91.
b. Liver and kidneys were weighed. Organ to body weight ratios were calculated.
c. The following tissues were preserved ln 10% formalin: treated and untreated skin, sternal bone marrow, liver, kidneys, thyroids, lung,heart, spleen, adrenals, mesenteric lymph nodes, urinary bladder, testes with epididymus, ovaries, sciatic nerve with muscle, spinal cord (2 levels), brain, pituitary, pancreas, trachea, esophagus, and stomach.
d. The organs that were processed histologically, including haematoxylin and eosin staining,and examined histomorphologically were: skin,kidneys, liver, sternal bone and spinal cord.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
All but one of the animals survived the test in generally good health. A male rat (2912), Group II-HCA, died on Day 14 after losing weight and refusing to drink water. Emaciation, starting Day 7, lethargy, and ptosis were also noted
Dermal irritation:
no effects observed
Description (incidence and severity):
All sites were normal at all times
Mortality:
mortality observed, treatment-related
Description (incidence):
All but one of the animals survived the test in generally good health. A male rat (2912), Group II-HCA, died on Day 14 after losing weight and refusing to drink water. Emaciation, starting Day 7, lethargy, and ptosis were also noted
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weights were generally comparable to control. In Group II, the rate of gain was very slightly less for females and slightly less for males. However, none of the differences was statistically significant.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
There were no remarkable differences from control.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Most parameters were comparable to control. There was a slight increase in alkaline phosphatase values in males of both treated groups, This was not noted in females. A relationship to treatment is considered to be questionable
Urinalysis findings:
no effects observed
Description (incidence and severity):
There was no remarkable difference from control
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There was no remarkable difference from control.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Necropsy findings in the rat which died, 2912M, Group II, HCA, were indicative of a lung infection. Other findings at sacrifice on Day 91, appeared to be spontaneous in nature and unrelated to treatment.

Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no significant lesions on treated or untreated skin. bone marrow or spinal cord. Findings in the liver and kidney were similar to lesions found in normal, non-experimental animals and were therefore considered to be unrelated to treatment
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Key result
Dose descriptor:
NOEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on lack of effects observed at the highest dose tested.
Key result
Critical effects observed:
no

Table 2. Clinical Chemistry - Males

Group

 

A.P. (IU/L)

I

Mean

23.0

S.D.

1.5

n

5

 

II

Mean

36.9*

S.D.

8.6

n

4

Conclusions:
One male rat treated with Hexyl Cinnamic Aldehyde died on Day 14. At necropsy, there was evidence of a lung infection. The death was not considered to be related to treatment. Of the other parameters evaluated, there was no evidence of toxicity induced by treatment with the test article. The NOEL for repeated dose toxicity via the dermal route was therefore considered to be 25 mg/Kg bw/day.
Executive summary:

In a key repeated dose dermal toxicity study, the test material (Hexyl Cinnamic Aldehyde in phenyl ethyl alcohol) at a dose of 25 mg/Kg was applied topically to the clipped backs of Sprague Dawley Albino rats daily for 90 days. There were five male and five female rats/group. A control group of five male and five female rats received Phenyl Ethyl Alcohol, 1 mL/Kg.

 

All rats were observed daily and skin reactions were recorded. Body weights were recorded weekly. At termination, selected haematology, clinical chemistry and urinalysis parameters were evaluated. All animals were examined grossly and liver and kidneys were weighed. Microscopic examination of the skin, liver, kidney and spinal cord was conducted.

 

One male rat treated with Hexyl Cinnamic Aldehyde died on Day 14. At necropsy, there was evidence of a lung infection. The death was not considered to be related to treatment. Of the other parameters evaluated, there was no evidence of toxicity induced by treatment with the test article.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report Date:
1981

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
5% Hexyl Cinnamic Aldehyde in phenyl ethyl alcohol
The specific gravity for all the compound was 1.00.
The test articles were stored for the duration of the study at ambient temperature and humidity.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Ace Animals, Inc., P.O. Box 122, Boyertown, PA
- Females (if applicable) nulliparous and non-pregnant: Not specified
- Age at study initiation: Not specified
- Weight at study initiation: 75-100g
- Fasting period before study: Not specified
- Housing: The animals were housed/caged (Rats will be individually housed in wire mesh cages in a temperature controlled room reserved exclusively for this test. Cages will meet the minimum size requirement specified in DHEU Publication No. (NIH) 77-23, entitled "Guide for the Care and Use of Laboratory Animals) for 3 days after receipt to facilitate adjustment to the watering system. Thereafter, they were individually housed.
- Diet (e.g. ad libitum): Purina Rat Mash (#5001) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: The animals were kept in a room reserved for this test from date of receipt until dosing was initiated. During this period, the rats were observed daily. Body weights where recorded.

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The animal room was temperature and humidity controlled
- Humidity (%): The animal room was temperature and humidity controlled
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): Light was automatically controlled on a 12 hour light/dark cycle.

IN-LIFE DATES: From: 1981-02-03 To: 1981-06-07

Administration / exposure

Type of coverage:
not specified
Vehicle:
other: Phenyl ether alcohol
Details on exposure:
The test articles were applied topically, 7 days/week for 90 days by syringe and ball-tipped needle. The backs were divided into a seven (7) area grid and a separate area was treated daily to minimize local irritation. The rats were clipped once a week or as needed. Dosage volumes were based on weekly body weights.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
7 days/week for 90 days
Frequency of treatment:
90 days
Doses / concentrations
Dose / conc.:
25 mg/kg bw/day (nominal)
No. of animals per sex per dose:
17/sex
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Not specified
- Rationale for animal assignment (if not random): Rats in good health were assigned to study groups by random numbers generated by a program in the TI 59 programmable calculator. Ear tags were secured after randomization.

Examinations

Observations and examinations performed and frequency:
The rats were observed daily for signs of toxicity, pharmacological effects and mortality
Body Weights were recorded weekly
Dermal observations were recorded daily.
Clinical studies: The following parameters were measured at termination in all rats
hematology: erythrocytes, hemoglobin, hematocrit and total and differential leucocytes.
clinical chemistry: glucose, blood urea nitrogen, cholesterol, total bilirubin, uric acid, lactic dehydrogenase, albumin total protein. calcium, phosphorus. alkaline phosphatase, serum glutamic-oxaloacetic transaminase and serumglutamic - pyruvic transaminase.
urinalysis: appearance, color, specific gravity, pH, protein, glucose, ketone, bilirubin, occult blood and microscopic examination of the sediment

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily. The rats were observed daily for signs of toxicity, pharmacological effects and mortality.

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION: No data
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Not specified
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: erythrocytes, hemoglobin, hematocrit and total and differential leucocytes.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Not specified
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: glucose, blood urea nitrogen, cholesterol, total bilirubin, uric acid, lactic dehydrogenase, albumin, total protein. calcium, phosphorus. alkaline phosphatase, serum glutamic-oxaloacetic transaminase and serum glutamic-pyruvic transaminase

URINALYSIS: Yes
- Time schedule for collection of urine: Not specified
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters examined: appearance, color, specific gravity, pH, protein, glucose, ketone, bilirubin, occult blood and microscopic examination of the sediment

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

a. Animals were anesthetised with ether and exsanquinated on Day 91.
b. Liver and kidneys were weighed. Organ to body weight ratios were calculated.
c. The following tissues were preserved ln 10% formalin: treated and untreated skin, sternal bone marrow, liver, kidneys, thyroids, lung,heart, spleen, adrenals, mesenteric lymph nodes, urinary bladder, testes with epididymus, ovaries, sciatic nerve with muscle, spinal cord (2 levels), brain, pituitary, pancreas, trachea, esophagus, and stomach.
d. The organs that were processed histologically, including haematoxylin and eosin staining,and examined histomorphologically were: skin,kidneys, liver, sternal bone and spinal cord.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
All but one of the animals survived the test in generally good health. A male rat (2912), Group II-HCA, died on Day 14 after losing weight and refusing to drink water. Emaciation, starting Day 7, lethargy, and ptosis were also noted
Dermal irritation:
no effects observed
Description (incidence and severity):
All sites were normal at all times
Mortality:
mortality observed, treatment-related
Description (incidence):
All but one of the animals survived the test in generally good health. A male rat (2912), Group II-HCA, died on Day 14 after losing weight and refusing to drink water. Emaciation, starting Day 7, lethargy, and ptosis were also noted
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weights were generally comparable to control. In Group II, the rate of gain was very slightly less for females and slightly less for males. However, none of the differences was statistically significant.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
There were no remarkable differences from control.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Most parameters were comparable to control. There was a slight increase in alkaline phosphatase values in males of both treated groups, This was not noted in females. A relationship to treatment is considered to be questionable
Urinalysis findings:
no effects observed
Description (incidence and severity):
There was no remarkable difference from control
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There was no remarkable difference from control.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Necropsy findings in the rat which died, 2912M, Group II, HCA, were indicative of a lung infection. Other findings at sacrifice on Day 91, appeared to be spontaneous in nature and unrelated to treatment.

Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no significant lesions on treated or untreated skin. bone marrow or spinal cord. Findings in the liver and kidney were similar to lesions found in normal, non-experimental animals and were therefore considered to be unrelated to treatment
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Effect levels

Key result
Dose descriptor:
NOEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on lack of effects observed at the highest dose tested.

Target system / organ toxicity

Key result
Critical effects observed:
no

Any other information on results incl. tables

Table 2. Clinical Chemistry - Males

Group

 

A.P. (IU/L)

I

Mean

23.0

S.D.

1.5

n

5

 

II

Mean

36.9*

S.D.

8.6

n

4

Applicant's summary and conclusion

Conclusions:
One male rat treated with Hexyl Cinnamic Aldehyde died on Day 14. At necropsy, there was evidence of a lung infection. The death was not considered to be related to treatment. Of the other parameters evaluated, there was no evidence of toxicity induced by treatment with the test article. The NOEL for repeated dose toxicity via the dermal route was therefore considered to be 25 mg/Kg bw/day.
Executive summary:

In a key repeated dose dermal toxicity study, the test material (Hexyl Cinnamic Aldehyde in phenyl ethyl alcohol) at a dose of 25 mg/Kg was applied topically to the clipped backs of Sprague Dawley Albino rats daily for 90 days. There were five male and five female rats/group. A control group of five male and five female rats received Phenyl Ethyl Alcohol, 1 mL/Kg.

 

All rats were observed daily and skin reactions were recorded. Body weights were recorded weekly. At termination, selected haematology, clinical chemistry and urinalysis parameters were evaluated. All animals were examined grossly and liver and kidneys were weighed. Microscopic examination of the skin, liver, kidney and spinal cord was conducted.

 

One male rat treated with Hexyl Cinnamic Aldehyde died on Day 14. At necropsy, there was evidence of a lung infection. The death was not considered to be related to treatment. Of the other parameters evaluated, there was no evidence of toxicity induced by treatment with the test article.