Registration Dossier

Administrative data

Description of key information

Skin Irritation:

The dermal irritation potential of test chemical was assessed in various experimental studies conducted on rabbits. Based on the available data for the key and supporting studies, it can be concluded that the test chemical is unable to cause skin irritation and thus considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Eye irritation:

An ocular irritation potential of target chemical was assessed in various experimental studies conducted on test animals. Based on the available data for the key and supporting studies, it can be concluded that the test chemical is not able to cause eye irritation and thus can be considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not classified (Not irritating)”.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: EWG Directive 84/449/EWG
Principles of method if other than guideline:
Pnimary skin irritation study with test chemical In rabbits (4-hour semi occlusive application)
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Remarks:
KFM (SPF quality)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG
CH 4414 Fuellinsdorf/ Switzerland
- Age at study initiation: 14-15 weeks
- Weight at study initiation: 2.3-2.7 kg
- Housing:Individually in stainless steel cage eqiipped with an automatic cleaning and drinking system.
- Diet (e.g. ad libitum): Pelleted stardand Kuba 341, Batch 17/86 rabbit maintenance diet
- Water (e.g. ad libitum):Community tap water from ltingen. ab Libitum.
- Acclimation period:Three days under trest conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22±3°C
- Humidity (%):relative humidity 40-70 %
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, music/light period.

IN-LIFE DATES: From: To: 8 August 1986 to 25 August 1986
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): No data


NEGATIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution): No data

POSITIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution):No data
Duration of treatment / exposure:
4 hours
Observation period:
The skin irritation was observed at 1, 24, 48 and 72 hrs and at 7 and 14 days intervals after removing the dressing.
Number of animals:
3 animals:
1 male
2 females
Details on study design:
TEST SITE
- Area of exposure: Dorsal area
- % coverage: 100 square cm (10cm x10cm)
- Type of wrap if used: 3.0cm x 3.0 cm patch of surgical gauze, gauze was covered with semi occlusive dressing, the dressing was anchored with elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing : Washing done with lukewarm water
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS : 1, 24, 48 and 72 hrs and at 7 and 14 days intervals after removing the dressing.

SCORING SYSTEM:
- Method of calculation: EEC Council Directive 83/467/EEC,
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 1 hr to 14 days
Score:
0.9
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
probability of mild irritation
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 1 hr to 14 days
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
probability of mild irritation
Irritant / corrosive response data:
Mild erythema was observed in all three animals but the signs were reversible within 14 days.
Other effects:
No data available.

Table 5: Primary skin irritation index

Selected observations

Mean cumulative

24 hrs

1.0

48 hrs

1.3

72 hrs

 1.3

7 days

0.7

14 days

0.0

 

Table 6: Mean scores per animal

Animal

Erythema

Edema

412: Male

1.2

0

413: Female

0.6

0

413: Female

0.8

0

Interpretation of results:
other: Not irritating
Conclusions:
In a skin irritation study, a maximum score of 0.9 out of 4 was observed. Slight irritant effects were observed which were fully reversible in 14 days. Therefore the test chemical can be considered to be not irriotant to the rabbit skin.
Executive summary:

A skin irritation study was performed on 1 male and 2 female New Zealand White KFM (SPF quality) rabbits. The test chemical was applied undiluted at a concentration of 0.5 ml to the intact shaved skin semiocclusively for four hours. The skin irritation was observed at 1, 24, 48 and 72 hrs and at 7 and 14 days intervals after removing the dressing. Body weight was measured at day 1 and at termination of the study. Rabbits were observed for mortality daily. All the observations were terminated 14 days after the application of the test chemical. Rabbits were killed by an intraveneous injection of T61 into the ear vein. The skin reaction like irritation score, colouration, corrosion and toxic symptoms waere assessed according to the OECD guideline 404. The test chemical showed primary irritation score of 0.9 when applied to the intact rabbit skin. Local signs consisted a grade 0.9 erythema and grade 0.0 edema. The skin reactions were reversible until day 14. Slight coloration was observed, no corrosive and toxic symptoms were observed in the test animals.

Based on obove observations and at a score of 0.9, the test chemical can be classified as "Not irritant" according to the CLP regulation.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: EWG Directive 84/449/EWG
Principles of method if other than guideline:
Determination of primary eye irritation study test chemical in rabbits.
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Remarks:
KFM (SPF quality)
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG
CH 4414 Fuellinsdorf/ Switzerland
- Age at study initiation: 14-15 weeks
- Weight at study initiation: 2.3-2.7 kg
- Housing:Individually in stainless steel cage eqiipped with an automatic cleaning and drinking system.
- Diet (e.g. ad libitum): Pelleted stardand Kuba 341, Batch 17/86 rabbit maintenance diet
- Water (e.g. ad libitum):Community tap water from ltingen. ab Libitum.
- Acclimation period:Three days under trest conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22±3°C
- Humidity (%):relative humidity 40-70 %
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, music/light period.

IN-LIFE DATES: From: To: 8 August 1986 to 25 August 1986
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 ml of test chemical was applied per animal.
Duration of treatment / exposure:
No data
Observation period (in vivo):
The eyes of each animal were examined 1, 24, 48 and 72 hours, after administration.
Number of animals or in vitro replicates:
3 animals
1 male and 2 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data available
- Time after start of exposure: No data available

SCORING SYSTEM: No data available

TOOL USED TO ASSESS SCORE: hand-slit lamp
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
1.1
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
No staining of the conjuctiva and cornea and no corrosion was observed in the rabbits.
Other effects:
- Lesions and clinical observations: No data
- Ophthalmoscopic findings: No data
- Histopathological findings: No data
- Effects of rinsing or washing: No data
- Other observations:No data

Table 3: Primary irritation score

Selected observations

Mean scores

1 hr

2.7

24 hrs

1.3

48 hrs

0.3

72 hrs

0.0

 

Table 4: Mean scores per animal

Animal & Sex

Cornea opacity

Iris

Conjunctivae

Redness

Chemosis

415 M

0.0

0.0

0.7

0.3

146 F

0.0

0.0

0.3

0.0

417 F

0.0

0.0

0.0

0.0

Selected observations for mean: 24 hrs, 48 hrs and 72 hrs

Interpretation of results:
other: Not irritating
Conclusions:
The test chemical was found to cause a primary irritation score of 1.1 when applied to the rabbit eye mucosa. No sever effects were observed in 72 hours. Thus, the test chemical was considered to be not Irritating to the eyes.
Executive summary:

An eye irritation study was performed on 1 male and 2 female New Zealand White KFM (SPF quality) rabbits. The test chemical was applied undiluted at a concentration of 0.1 ml in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control. The eyes of each animal were examined 1, 24, 48 and 72 hours, after administration. Body weight was measured at day 1 and at termination of the study. Rabbits were observed for mortality daily. Rabbits were killed by an intraveneous injection of T61 into the ear vein. The irritation was assessed according to the OECD Guidelines for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion". Slight coloration was observed, no corrosive and toxic symptoms were observed in the test animals. Based on obove observations and a primary irritation score of 1.1, the test chemical can be classified as "Not irritant" to the rabbit eye according to the CLP regulation.

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

Additional information

Skin Irritation (In-vivo):

Various studies were performed on rabbits and rats to assess the dermal irritation potential of test chemical which have been summarized as follows:

A skin irritation study was performed on 1 male and 2 female New Zealand White KFM (SPF quality) rabbits. The test chemical was applied undiluted at a concentration of 0.5 ml to the intact shaved skin semiocclusively for four hours. The skin irritation was observed at 1, 24, 48 and 72 hrs and at 7 and 14 days intervals after removing the dressing. Body weight was measured at day 1 and at termination of the study. Rabbits were observed for mortality daily. All the observations were terminated 14 days after the application of the test chemical. Rabbits were killed by an intraveneous injection of T61 into the ear vein. The skin reaction like irritation score, colouration, corrosion and toxic symptoms waere assessed according to the OECD guideline 404. The test chemical showed primary irritation score of 0.9 when applied to the intact rabbit skin. Local signs consisted a grade 0.9 erythema and grade 0.0 edema. The skin reactions were reversible until day 14. Slight coloration was observed, no corrosive and toxic symptoms were observed in the test animals.

Based on obove observations and at a score of 0.9, the test chemical can be classified as "Not irritant" according to the CLP regulation.

Another skin irritation study was carried out according to OECD 404 “SKIN IRRITATION” to evaluate the irritation potential of the test chemical. 0.5 ml of test chemical was applied to the flank of 3 rabbits semi occlusively. Observations were made at 24, 48 and 72 hrs after application of test chemical. The scoring was done according to the Draize method. Erythema was observed in all 3 animals, edema was not observed. The primary irritation score of test chemical was 0.89. Based on the classification of chemicals, the test chemical can be considered as "Not irritating" to rabbit skin.

All these studies lead to a conclusion that Test chemical is indeed not irritating to skin. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

Similarly, another skin irritation study was carried out according to OECD 404 “SKIN IRRITATION” to evaluate the irritation potential of the test chemical. 0.5 ml of test chemical was applied to the flank of 3 rabbits semi occlusively. Observations were made at 24, 48 and 72 hrs after application of test chemical. The scoring was done according to the Draize method. No erythema and edema was observed. The primary irritation score of test chemical was 0. Based on the classification of chemicals, the test chemical can be considered as "Not irritating" to rabbit skin.

Skin Irritation (In-vitro):

The dermal irritation potential of test article was determined according to the OECD 439 test guideline for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met and passed the acceptance of criteria. The Mean % tissue viability compared to negative control (n=3) of the test substance was determined to be 4.8%. Hence, under the current experimental test conditions it was concluded that test substance was considered to be irritating to human skin.

Though the in vitro results for the target indicates that test chemical can be irritating to skin, but the estimated in-vivo results imply that test chemical is not irritating to skin. These results are aptly supported by the results of rabbit study for the test chemical.

Thus, based on above observations, the test chemical be determined as "Not irritating" to skin of rabbits.

Eye Irritation (In-vivo):

Various studies were performed on test animals to assess the ocular irritation potential of test chemical which have been summarized as follows:

An eye irritation study was performed on 1 male and 2 female New Zealand White KFM (SPF quality) rabbits. The test chemical was applied undiluted at a concentration of 0.1 ml in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control. The eyes of each animal were examined 1, 24, 48 and 72 hours, after administration. Body weight was measured at day 1 and at termination of the study. Rabbits were observed for mortality daily. Rabbits were killed by an intraveneous injection of T61 into the ear vein. The irritation was assessed according to the OECD Guidelines for testing of Chemicals, Section 4, number 405 "Acute Eye Irritation/Corrosion". Slight coloration was observed, no corrosive and toxic symptoms were observed in the test animals. Based on obove observations and a primary irritation score of 1.1, the test chemical can be classified as "Not irritant" to the rabbit eye according to the CLP regulation.

Another eye irritation study was carried out according to OECD 405 “EYE IRRITATION” to evaluate the irritation potential of the test chemical. 0.1 ml of test chemical was instilled into the conjunctival sac of 3 New Zealand White rabbits. Observations were made after 1 hour, 4 hours and then 1, 2, 3 days after instillation of test chemical. The scoring was done according to the Draize method. Conjunctival redness and chemosis was observed in all 3 animals after 1 day of observation. The MMAS (Maximum Modified Average Score) of test chemical was 4.0 after 1 day of observation. Based on the classification of chemicals according to MMAS scores, the test chemical can be considered as "Not irritating" to rabbit eyes.

Similarly another eye irritation test was conducted on rabbits according to Directive 84/449/EEC, B.5 "Acute toxicity (eye irritation)" to evaluate the irritation potential of test material. Three rabbits were each administered a single ocular dose of 0.1 ml test substance and observed over a 7 day period. Effects were temporary corneal opacity (one animal), transient dulling of the cornea (1 animal), transient iridial inflammation (one animal) and slight to well defined conjunctival irritation. All reactions had resolved in 7 days. Hence, the test chemical can be considered as "Not irritating" to rabbit eyes.

Eye Irritation (In-vitro):

The ocular irritation potential of test article was determined according to the OECD 492 test guideline for this study. The MatTek EpiOcular™ model was used to assess the potential ocular irritation of the test articles by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to liquid test articles and controls for ~30 minutes, followed by a ~12 minute post-soak and approximately 2 hour recovery after the post-soak. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met, passing the acceptance criteria. The mean % tissue viability of test substance was determined to be 19.7%. Hence, under the experimental test conditions it was concluded that test substance was considered to be irritating to the human eyes and can thus be classified as ‘’Irritating to eyes in Category 2" as per CLP Regulation.

Though the in vitro results for the target indicates that test chemical can be irritating to eye, but the estimated in-vivo results imply that test chemical is not irritating to eye. These results are aptly supported by the results of rabbit study for the rtest chemical.


The results obtained from these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “ Not classified (not irritating to eyes) based on GHS criteria”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical were observed in various studies. The results obtained from these studies indicates that the chemical is unlikely to cause skin irritation and eye irritation. Hence the test chemical can be classified under the category “Not Classified” for skin and eye irritation as per CLP.