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Administrative data

Description of key information

In an in vitro skin irritation study, performed according to OECD/EC test guidelines, the substance was found to be irritant to the skin.  In an in vitro skin corrosion study, performed according to OECD/EC test guidelines, the substance was found not to be corrosive to the skin.  
In an eye irritation study with rabbits, performed according to OECD/EC test guidelines, limited irritation was observed (not sufficient for classification).

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
19NOV2012 to 10DEC2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
July 2010
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
March 2003
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
- Source: Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: Animals were at least 6 weeks old.
- Weight at study initiation: Body weights were between 1646 and 2284 g.
- Housing: Individually housed in cages with perforated floors.
- Diet: Free access to pelleted diet for rabbits (Global Diet 2030 Harlan Teklad, Italy). Hay and wooden sticks were available during the study period.
- Water: Free access to tap water.
- Acclimatization period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 19NOV2012 to 10DEC2012
Vehicle:
unchanged (no vehicle)
Controls:
other: One eye of each animal remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL- Amount applied (weight with unit): 49.8 mg (range 49.6 – 50.3 mg), a volume of approximately 0.1 mL.
Duration of treatment / exposure:
Single instillation on Day 1.
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance.
Number of animals or in vitro replicates:
3 males.
Details on study design:
STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 14 days later, after considering the degree of eye irritation observed in the first animal.

TREATMENT
Animals were treated by instillation of the test substance in the conjunctival sac of one of the eyes 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 substance. The other eye remained untreated and served as the reference control.

Immediately after the 24-hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

REMOVAL OF TEST SUBSTANCE
-Washing: No

OBSERVATIONS (beschrijf beperkt)
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Prior to instillation and after the final observation.
- Necropsy: No necropsy was performed according to protocol.
- Irritation: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded. The irritation was assessed according to OECD 405.
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
mean
Remarks:
of three animals
Time point:
other: 24, 48 and 72 hrs.
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
of three animals
Time point:
other: 24, 48 and 72 hrs.
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
mean
Remarks:
of three animals
Time point:
other: 24, 48 and 72 hrs.
Score:
1.1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of three animals
Time point:
other: 24, 48 and 72 hrs.
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
Instillation of approximately 50 mg (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in corneal injury consisting of slight dulling of the normal luster, noted in one animal on Day 1. Treatment of the eye with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage in any of the rabbits. Iridial irritation grade 1 was observed in one animal and resolved within 24 hours. Irritation of the conjunctivae was seen in all animals and consisted of redness, this had completely resolved in all animals within 7 days. All animals showed chemosis and discharge at 24 hrs after instillation, this had resolved after 48 hours.
Other effects:
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.
No changes in body weight gain occurred.
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an eye irritation study with rabbits, performed according to OECD/EC test guidelines, limited irritation was observed, not sufficient for classification.
Executive summary:

An eye irritation study was performed according to OECD/EC test guidelines and GLP principles. Approximately 50 mg (a volume of approximately 0.1 mL) was instilled into one eye of each of three rabbits. In one animal on Day 1, the corneal injury consisted of slight dulling of the normal luster. Redness of conjunctivae and chemosis was noted for all animals (mean 1.1 and 0.3 resp. at 24, 48 and 72 hrs), which had completely resolved after 7 days. Treatment of the eye with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage. No systemic toxicity, changes in body weight gain or mortaliy occurred.

According to these data, the test substance is not irritating to the eyes and is not classified for eye irritation.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In an in vitro skin irritation test using a human skin model ( EPISKIN Standard Model), the influence of TMAC on the viability of human skin was tested. Survival of unexposed skin was set at 100%, the positive control had a mean cell viability of 5% whereas the test substance showed cell viability of 28%. Since the mean relative tissue viability after exposure to the test substance was below 50%, it can be concluded that the test substance is irritating in the in vitro skin irritation test.

In an in vitro skin corrosion test using a human skin model (EpiDerm Skin Model), the influence of TMAC on the viability of human skin was tested. Survival of unexposed skin was set at 100%, the positive control had a mean cell viability of 26% and 10% after resp. 3 minutes or 1 hour exposure whereas the test substance showed a cell viability of 98% and 92%, resp. Since the mean relative tissue viability after exposure to the test substance was above 50% or 15% after resp. 3 minutes or 1 hour exposure, it can be concluded that the test substance is not corrosive in the in vitro skin corrosion test.

An eye irritation study was performed according to OECD/EC test guidelines and GLP principles. Approximately 50 mg (a volume of approximately 0.1 mL) was instilled into one eye of each of three rabbits. In one animal on Day 1, the corneal injury consisted of slight dulling of the normal luster. Redness of conjunctivae and chemosis was noted for all animals (mean 1.1 and 0.3 resp. at 24, 48 and 72 hrs), which had completely resolved after 7 days. Treatment of the eye with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage. No systemic toxicity, changes in body weight gain or mortaliy occurred. Based on these data, TMAC is not considered to be irritating to the eye.


Justification for selection of skin irritation / corrosion endpoint:
Two studies were selected as key studies: an in vitro skin irritation study (WIL 2012) and an in vitro skin corrosion study (WIL 2012). Both studies are needed for the endpoint conclusion. Both studies were conducted according to OECD guideline and GLP principles (both reliability 1).

Justification for selection of eye irritation endpoint:
One in vivo study available with reliability 1. The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

According to the available data, the test substance is irritating to the skin, but not corrosive and is classified according to CLP Regulation (EC) No. 1272/2008 with Cat.2. The substance was found not to be irritating to the eyes and thus does not have to be classified for eye irritation according to CLP Regulation (EC) No. 1272/2008.