Registration Dossier

Administrative data

Description of key information

One skin and one eye irritation assay (in vivo)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP/Guidline Study
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: Annex V
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
other: MAFF 59 NohSan No. 4200
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products Inc., Denver, PA
- Age at study initiation: Young adult
- Weight at study initiation: 2622 to 2654 grams
- Housing: Individual suspended wire-mesh cages.
- Diet (e.g. ad libitum): PMI Nutrition International, Inc. Certified Rabbit LabDiet® 5322 was offered at approximately 150 g per day during the study.
- Water (e.g. ad libitum): municipal water ad libitum
- Acclimation period: The animals were acclimated to laboratory conditions for a minimum of seven days prior to initiation of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature: 66-71°F
- Humidity (%): 30-68%
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES: From: 2001-02-14 to 2001-02-23
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 ml
Duration of treatment / exposure:
4 h
Observation period:
Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes adn 24,48 and 72 hours after patch removal and daily through study day 9, if irritation persisited.
Number of animals:
3
Details on study design:
There was one group of three albino rabbits that received a single, four-hour, semi-occluded exposure. Each 0.5-ml dose of the test article was applied to the clipped, unabraded skin. At completion of exposure, the bandages were removed and the sites washed.

Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes adn 24,48 and 72 hours after patch removal and daily through study day 9, if irritation persisited.

Body weights were obtained and recorded on study day 0 (initiation) and at each rabbit's termination from the study.

Upon termination, the rabbits were euthanized by intravenous injection of euthanasia solution and discarded.
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
1.3
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
2
Irritation parameter:
erythema score
Max. score:
2
Remarks on result:
other: Max. duration: 96 d; Max. value at end of observation period: 0 (related to all animals)
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
0
Irritation parameter:
edema score
Max. score:
0
Remarks on result:
other: Max. duration: 0 d; Max. value at end of observation period: 0 (related to all animals)
Irritant / corrosive response data:
Reversibility of any observed effect: Changes fully reversible within 9 days
Other effects:
Grade 2 erythema was still apparent in one animal at 96 hour s.

There were no deaths during the study.

There were no remarkable body weight changes noted during the study.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
All animals were noted with very slight to slight erythema which completely subsided by termination (study day 9). Two animals were noted with desquamation, which persisted through each animal’s termination (study days 7 or 9). There was no edema, nor were there any other cutaneous findings.
Executive summary:

The primary skin irritation potential of Consolidated Cyclohexane Dicarboxaldehydes was evaluated in this study with New Zealand White rabbits. There was one group of three albino rabbits that received a single, four-hour semi-occluded exposure. Each 0.5 -

ml dose of the test article was applied to the clipped unabraded skin. At completion of exposure, the bandages were removed and the sites washed. Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes and 24, 48, and 72 hours after patch removal and daily through study day 9, if irritation persisted. All animals were noted with very slight to slight erythema which completely subsided by termination (study day 9). Two animals were noted with desquamation, which persisted through each animal’s termination (study days 7 or 9). There was no edema, nor were there any other cutaneous findings. The primary Irritation Index (PII)( was 1.5. The test article, Consolidated Cyclohexane Dicarboxaldehydes, received a descriptive rating classification of slightly irritating.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2001-02-15 to 2001-03-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP/Guideline Study
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
other: Annex V
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF No. 6283
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products Inc., Denver, PA
- Age at study initiation: Young adult
- Weight at study initiation: 2602 to 2757 grams
- Housing: Individual suspended wire-mesh cages.
- Diet (e.g. ad libitum): PMI Nutrition International, Inc. Certified Rabbit LabDiet® 5322 was offered at approximately 150 g per day during the study.
- Water (e.g. ad libitum): municipal water ad libitum
- Acclimation period: The animals were acclimated to laboratory conditions for a minimum of seven days prior to initiation of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature: 63-72°F
- Humidity (%): 30-78%
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES: From: 2001-02-15 to 2001-03-15
Vehicle:
unchanged (no vehicle)
Controls:
other: Left eye served as control for each animal
Amount / concentration applied:
.1 ML
Duration of treatment / exposure:
single, unwashed exposure
Observation period (in vivo):
Both eyes of all rabbits were examined macroscopically for ocular irritation in
accordance with the method of Draize (Appendix A) at approximately one, 24, 48 and
72 hours after dosing and on study days 4, 7, 10, 17 and 21, if irritation persisted. A
direct ophthalmoscope was used during these observations to examine the corneal
tissue. In addition, both eyes were further examined at 24 hours and at all subsequent
observations with sodium fluorescein.
Number of animals or in vitro replicates:
3
Details on study design:
There was one group of three albino rabbits that received a single, unwashed
exposure. Each 0.1-ml dose of the test article was instilled into the lower
conjunctival sac of the right eye. The eyelid was held closed for approximately one
second and released. The left eye was manipulated in a similar manner as the right
eye and served as a contralateral control.
The eyes were examined for ocular reactions in accordance with the method of Draize
(Appendix A) at approximately one, 24, 48 and 72 hours after dosing and on study
days 4, 7, 10, 17 and 21, if irritation persisted. Sodium fluorescein was used to aid in
revealing possible corneal damage at 24 hours and at all subsequent observations.

Body weights were obtained and recorded on study day 0 (initiation) and at each
rabbit’s termination from the study.

After study termination, the rabbits were euthanized by intravenous injection of
euthanasia solution and discarded.
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
2
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
2.7
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
2
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Max. score:
3
Remarks on result:
other: Max. duration: h; Max. value at end of observation period: 1 (related to all animals)
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
3
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
3.7
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
3
Irritation parameter:
chemosis score
Max. score:
4
Remarks on result:
other: Max. duration: h; Max. value at end of observation period: 0 (related to all animals)
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
cornea opacity score
Max. score:
2
Remarks on result:
other: Max. duration: h; Max. value at end of observation period: 2 (related to all animals)
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
(mean score 1)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
(mean score 2)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
(mean score 3)
Time point:
other: overall at 24, 48 and 72 h
Score:
1
Irritation parameter:
iris score
Max. score:
1
Remarks on result:
other: Max. duration: 96 h; Max. value at end of observation period: 0 (related to all animals)
Irritant / corrosive response data:
Reversibility of any observed effect: Changes not fully reversible within 21 days

No remarkable body weight changes were noted during the study.

There were no deaths during the study.

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
All animals were noted with positive cornea1 (grade l-2 for opacity, grade 4 for area), iridal (grade 1) and conjunctival (grade 2-3 for redness, grade 3-4 for chemosis) findings. Irritation for two animals completely subsided by study days 10 or 17. Corneal and conjunctival findings were still present at study termination for one animal. Additionally, this animal was observed with cornea1 neovascularization on study days 10 and 17. The Maximum Average Score for Consolidated Cyclohexane Dicarboxaldehydes was 41.0 at 24 hours post-instillation.
Executive summary:

The primary ocular irritation potential of Consolidated Cyclohexane Dicarboxaldehydes was evaluated in this study with New Zealand White rabbits. There was one group of three albino rabbits that received a single, unwashed exposure. Each 0.1- ml dose of the test article was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately one second and released. The left eye was manipulated in a similar manner as the right eye and served as a contralateral control. The eyes were examined for ocular reactions in accordance with the method of Draize at approximately one, 24, 48 and 72 hours after dosing and on study days 4, 7, 10, 17 and 21, if irritation persisted. Sodium fluorescein was used to aid in revealing possible corneal damage at

24 hours and at all subsequent observations. All animals were noted with positive corneal (grade 1-2 for opacity, grade 4 for area), iridal (grade 1) and conjunctival (grade 2-3 for redness, grade 3-4 for chemosis) findings. Irritation for two animals completely subsided by study days 10 or 17. Corneal and conjunctival findings were still present at study termination for one animal. Additionally, this animal was observed with corneal neovascularization on study days 10 and 17. The Maximum Average Score for Consolidated Cyclohexane Dicarboxaldehydes was 41.0 at 24 hours post-instillation.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Skin irritation

The primary skin irritation potential of Consolidated Cyclohexane Dicarboxaldehydes was evaluated in this study with 3 New Zealand White rabbits. All animals were noted with very slight to slight erythema which completely subsided by termination (study day 9). Two animals were noted with desquamation, which persisted through each animal’s termination (study days 7 or 9). There was no edema, nor were there any other cutaneous findings. The primary Irritation Index (PII) was 1.5. The test article, Consolidated Cyclohexane Dicarboxaldehydes, received a descriptive rating classification of slightly irritating.

Eye irritation:

The primary ocular irritation potential of Consolidated Cyclohexane Dicarboxaldehydes was evaluated in this study with 3 New Zealand White rabbits. All animals were noted with positive corneal (grade 1-2 for opacity, grade 4 for area), iridal (grade 1) and conjunctival (grade 2-3 for redness, grade 3-4 for chemosis) findings. Irritation for two animals completely subsided by study days 10 or 17. Corneal and conjunctival findings were still present at study termination (day 21) for one animal. Additionally, this animal was observed with corneal neovascularization on study days 10 and 17.

Respiratory irritation:

A 2 week, repeated dose (6 hours per day, 5 days per week) inhalation range finding study was performed using F344 rats (5 male and 5 female per dose group). The dose levels chosen were 0, 4.4, 26.8, or 93.2 mg1,3- and 1,4-cyclohexanedicarboxaldehyde/m3air. The main findings of this study are discussed in section '7.5 - Repeated dose toxicity'. With respect to irritation to the respiratory tract,

lesions of the respiratory tract occurred inthe anterior nasal cavity of animals exposed to 93.2 and 26.8 mg/m3and cranial larynx of animals exposed to 93.2 mg/m3. The nasal cavity of male and female rats exposed to 26.8 or93.2 mg/m3had erosive/ulcerative, metaplastic/hyperplastic, and inflammatory changes to the squamous, respiratory, transitional epithelia, and hypertrophy of mucous cells lining the nasopharynx. The cranial larynx of male and female rats exposed to 93.1 mg/m3had very slight squamous metaplasia of the surface epithelium overlying the cartilage at the base of the epiglottis.

Based on these findings, this substance can be considered to be irritating to the upper respiratory tract, and the NOEL for this effect is the low dose of 4.4 mg/m3.

In addition to this study, data are available from an RD50 assessment in mice that demonstrated a dose dependant decrease in respiration rate following exposure to the test material (refer to section 7.12). The RD50 of inhaled 1,3- and 1,4-Cyclohexanedicarboxaldehyde was therefore determined to be 15.9 ppm, with a 95% confidence interval of 10.9 - 23.1 ppm.Taken together with the findings from the 14 day inhalation study, this substance can be considered as irritating to the respiratory tract.


Justification for selection of skin irritation / corrosion endpoint:
GLP guideline study

Justification for selection of eye irritation endpoint:
GLP guideline study

Effects on eye irritation: highly irritating

Effects on respiratory irritation: irritating

Justification for classification or non-classification

This substance does not meet the criteria for classification as irritating to the skin. However, one grade of this substance contains up to 10% n-heptane and as such requires classification as irritating to the skin (see section 2.1).

Based on the persistence of eye irritation in one animal through to the end of the eye irritation study this substance meets the criteria for classification as Category 1, irreversible effects on the eye, according to CLP, and R41, risk of serious damage to the eyes according to the DSD.

Based on the evidence of irritation to the respiratory tract in the 2 -week inhalation study and RD50 assay, this substance meets the criteria for classification for respiratory tract irritation, STOT-SE Cat 3.