Registration Dossier

Administrative data

Description of key information

Skin irritation/corrosion: not irritating (OECD 404, GLP);  
Eye irritation: not irritating (OECD 405, GLP)

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:
2015-12-13 to 2016-01-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
2015-07-28
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 15 - 25 °C, tightly closed
Species:
rabbit
Strain:
other: Zika
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Kaninchenbetrieb Kock, Warnkenhagen, Germany
- Age at study initiation: 13 (one rabbit) and 18 weeks old (two rabbits)
- Weight at study initiation: 3.41 to 3.96 kg
- Housing: single-caged; during the exposure time the rabbits were kept in a restrainer avoiding ingestion or inhalation of the test substance and avoiding access to the patch by the animal.
- Diet: conventional laboratory diet (a half-and-half blend of "Holstenstolz Kaninchenverbrauchsfutter Z, Type 038" and "Rabbit maintenance, MuesliMash")
- Water (ad libitum): tap water (drinking quality)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 °C (± 3 °C)
- Relative humidity: 30 - 70 %
- Air changes: 15/hour
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of test substance moistened with vehicle

VEHICLE
- Amount(s) applied (volume or weight with unit): 500 µL


Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3 female rabbits
Details on study design:
INITIAL AND CONFIRMATORY TEST
Prior to the initial test the pH of the test substance in water (saturated, sedimented suspension, 5.55 % w/v) was determined (pH = 7.46).
The in vivo test was performed initially using one animal. The first patch was removed after three minutes. No serious skin reaction was observed, therefore a second patch was applied at a different site and removed after one hour. No serious skin reaction was observed, therefore a third patch was applied at a different site and removed after four hours.
Because of a negative response after the initial test, two additional animals were exposed simultaneously. A single patch was applied to both animals for 4 hours.

TEST SITE
- Area of exposure: dorsal area of the trunk (area: approx. 6 cm²)
- Type of wrap if used: test substance was applied to the clipped skin and covered with a gauze patch (compress for operation, 8-fold), which was held in place with a non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing: residual test substance was removed, using water without altering the existing response or the integrity of the epidermis
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS (4 hour application time)
immediately (initial test only) as well as 1, 24, 48, and 72 hours after removal of the patch

SCORING SYSTEM: according to the Draize scale

Irritation parameter:
erythema score
Basis:
mean
Remarks:
amimal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Remarks:
aninmal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritant / corrosive response data:
No evidence of skin irritation/corrosion was noted during the study.
Other effects:
- Other adverse systemic effects: at the daily observation the animals did not show any other visible clinical symptoms.
Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not irritating to the skin based on an in vivo study (OECD 404).
According to Regulation (EC) No 1272/2008 and subsequent adaptations, the substance does not require classification as skin irritant.
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:
2015-12-15 to 2016-02-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2012-10-02
Deviations:
yes
Remarks:
application time of fluorescein after test item application was not stated
GLP compliance:
yes
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 15 - 25 °C, tightly closed
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test material was ground to a fine dust before application.
Species:
rabbit
Strain:
other: Zika
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kaninchenbetrieb Kock, Warnkenhagen, Germany
- Age at study initiation: 14 (one rabbit) and 18 weeks old (two rabbits)
- Weight at study initiation: 3.24 to 4.87 kg
- Housing: single-caged
- Diet (ad libitum): conventional laboratory diet (a half-and-half blend of "Holstenstolz Kaninchenverbrauchsfutter Z, Type 038" and "Rabbit maintenance, MuesliMash")
- Water (ad libitum): tap water (drinking quality)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 °C (± 3 °C)
- Relative humidity: 30 - 70 %
- Air changes: 15/hour
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg of the test substance
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
1, 24, 48 and 72 hours up to 5 (confirmatory test) or 9 days (initial test)
Number of animals or in vitro replicates:
3 female rabbits
Details on study design:
INITIAL AND CONFIRMATORY TEST
The in vivo test was performed initially using one animal. Prior to the initial test the pH of the test substance in water (saturated, sedimented suspension, 5.55 % w/v)) was determined (pH = 7.46).
Becasue of lack of corrosive or severe irritant reactions after the initial test, two additional animals were exposed.

USE OF TOPICAL ANAESTHETICS AND SYSTEMIC ANALGESICS
Approx. 60 minutes prior to test substance application, Alvegesic vet. (concentration: 10 mg/mL) was applied (application volume: 0.3 mg/kg bw).
Approx. five minutes prior to test substance application, one drop of a topical ocular anaesthetic (Proparakain-POS 0.5 %) was applied to each eye.
Four to six hours after test substance application, Alvegesic vet. (concentration: 10 mg/mL) subcutaneous was administered to provide a continued therapeutic level of systemic analgesia. An additional application was not necessary.

REMOVAL OF TEST SUBSTANCE
- Washing: test substance had not been removed from the eye of the test animals by physiological mechanisms at the first observation time, therefore the eyes were rinsed with physiological saline
- Time after start of exposure: 1 hour after treatment

SCORING SYSTEM: according to the Draize scale

TOOL USED TO ASSESS SCORE: slit lamp biomicroscope and fluorescein staining

CLINICAL OBSERVATIONS:
Test animals were evaluated for the entire duration of the study for clinical signs of pain and/or distress at least twice daily, with a minimum of 6 hours between observations, or more often if necessary.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
3
Remarks on result:
other: moderate conjunctivae redness was observed at the 1 hour observation.
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: Slight chemosis was observed at the 1 hour observation.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
3
Remarks on result:
other: severe conjunctivae redness was observed at the 1 hour observation.
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: Slight chemosis was observed at the 1 hour observation.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: moderate conjunctivae redness was observed at the 1 hour observation.
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: slight chemosis was observed at the 1 hour observation.
Irritant / corrosive response data:
Initial test (one rabbit): reversible signs of eye reactions (conjunctivae redness, chemosis) to the test substance were observed at 1 hour (also at 4 hours and 7 hours) after application.
Confirmatory test (two rabbits): reversible signs of eye reactions (conjunctivae redness, chemosis) to the test substance were observed at 1 hour (also 5 hours and 8 hours) and at 24 hours (one rabbit only) after application.
Other effects:
- Lesions and clinical observations: at the daily observations the animals did not show any other visible clinical symptoms.
Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not irritating to the eyes.
According to the EC Regulation No. 1272/2008 and subsequent adaptations, the substance is not classified as an eye irritant.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

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

Additional information

Skin irritation

The substance was not observed to be irritating to the skin in a reliable in vivo skin irritation study according to OECD 404.

Eye irritation

The substance was not observed to be irritating to the eyes in a reliable in vivo eye irritation study according to OECD 405.

Justification for classification or non-classification

Skin irritation

The substance does not possess a skin irritating potential based on an in vivo OECD 404 (2015) test and does not require classification as skin irritating according to Regulation (EC) No 1272/2008 and its subsequent adaptations.

Eye irritation

The substance does not possess an eye irritating potential based on an in vivo OECD 405 (2012) test and does not require classification as eye irritating according to Regulation (EC) No 1272/2008 and its subsequent adaptations.

Respiratory tract irritation

No signs of respiratory tract irritation were observed in an acute inhalation toxicity study in rats, up to the maximum attainable concentration. No classification required as respiratory irritatant according to regulation (EC) 1272/2008.