Registration Dossier

Administrative data

Description of key information

Following exposure with, the mean cell viability was 105% compared to the negative control and therefore non-irritant. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

In this in vitro skin irritation test in the EPISKIN model with the results indicated that the test item is Non Irritant (NI).

The test item, applied to rabbit eye mucosa, caused conjunctival redness and transient discharge. The effects were fully reversible within 48 hours.

The animal individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

chemosis : 0.00, 0.00

discharge : 0.00, 0.00

redness : 0.33, 0.33

cornea opacity : 0.00, 0.00

iris : 0.00, 0.00

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25 September 2013 - 27 September 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guidelines and GLP.
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. certificate)
Species:
human
Strain:
other: EPISKIN-SM
Irritation / corrosion parameter:
other: other: cell viability
Run / experiment:
mean
Value:
105
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
Basis: mean. Remarks: The OD value for the test item treated skin showed a viability of 105%.. (migrated information)
Other effects / acceptance of results:
For results of the optical density (OD) measured at 540 nm of each extract and the calculated % viability of the cells see attached table.
Colouring potential of test substances
As the test item is coloured, one additional test item-treated tissue was used for the non specific OD evaluation. Optical density (measured at 540 nm) of this tissue was determined as 0.034, Non Specific Colour % was calculated as 4.7%. Therefore additional data calculation was not necessary.

No colour change was observed after three hours of incubation of the test item in MTT solution. The test material did not interact with the MTT, therefore additional controls and data calculations were not necessary. A false estimation of viability can be precluded.
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Following exposure with PREPOLYMER D, the mean cell viability was 105% compared to the negative control and therefore non-irritant. All validity criteria were within acceptable limits and therefore the study can be considered as valid.
In this in vitro skin irritation test in the EPISKIN model with PREPOLYMER D the results indicated that the test item is Non Irritant (NI).
Executive summary:

The reconstructed human epidermis model EPISKIN-SM is designed to predict and classify the skin irritant potential of chemicals, by measuring its cytotoxic effect, as reflected in the MTT assay, on the EPISKIN reconstituted human epidermis. This method is approved by international regulatory agencies as a replacement for the identification of irritants / corrosives in the in vivo Rabbit skin assay (OECD 404).

Disks of EPISKIN (three units / chemical) were treated with PREPOLYMER D and incubated for 15 minutes at room temperature. Exposure of test material was terminated by rinsing with PBS. Epidermis units were then incubated at 37°C for 42 hours in an incubator with 5% CO2. The viability of the epidermis on each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37°C in 5% CO2 protected from light. The formazan extract in acidified isopropanol was then spectrophotometrically evaluated for optical density (OD) and quantified.

SDS 5% and PBS treated epidermis were used as positive and negative controls, respectively. An additional disk was used to provide an estimate of colour contribution from the test item. For each treated tissue viability was expressed as a % relative to negative control. If the mean relative viability after 15 minutes exposure and 42 hours post incubation is less or equal (≤) to 50% of the negative control, the test substance is considered to be irritant to skin.

Following exposure with PREPOLYMER D, the mean cell viability was 105% compared to the negative control and therefore non- irritant. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

In this in vitro skin irritation test in the EPISKIN model with PREPOLYMER D the results indicated that the test item is Non Irritant (NI).

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:
17 September 2013 - 22 September 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guidelines and GLP.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Species and strain: New Zealand White rabbits
Source: S&K-LAP Kft.
2173 Kartal, Császár út 135, Hungary
Justification of strain: The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined prior to starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
Number of animals: 2 animals
Age of animals at treatment: ~12 weeks old (adult)
Sex: Male
Body weight range on the day of treatment: 3240 g and 3410 g
before euthanasia: 3291 g and 3477 g
Date of receipt: 30 August 2013
Acclimatization time: at least 18 days
Animal identification: The individual identification was by engraved ear tag. The cages were marked with individual identity cards with information about study code, sex, dose, cage number and individual animal number.

Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 610
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
(and during of the analgesic/anaesthetic treatment)
Temperature
during the study: 20 ± 3°C
Relative humidity
during the study: 38 - 78 %
Housing/Enrichment: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages
Ventilation: 15-20 air exchanges/hour
The temperature and relative humidity values were measured continuously. The measured range was checked at least daily during the acclimatisation and experimental phases.
Vehicle:
unchanged (no vehicle)
Controls:
other: The untreated right eye served as control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
Duration of treatment / exposure:
72 hours
Observation period (in vivo):
The eyes were examined at 1, 24, 48 and 72 hours after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes.
Number of animals or in vitro replicates:
2
Details on study design:
The test substance was not rinsed from the eyes.

SCORING SYSTEM: The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (02 October 2012)

TOOL USED TO ASSESS SCORE: none
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
other: 24-48-72 hours
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24-48-72 hours
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24-48-72 hours
Score:
0
Max. score:
2
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24-48-72 hours
Score:
0
Max. score:
3
Irritation parameter:
other: discharge
Basis:
mean
Time point:
other: 24-48-72 hours
Score:
0
Max. score:
3
Irritant / corrosive response data:
see attached table
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item, applied to rabbit eye mucosa, caused conjunctival redness and transient discharge. The effects were fully reversible within 48 hours. The substance is not classified as irritating based on these results.
Executive summary:

An acute eye irritation study of the test item was performed in New Zealand White rabbits.

The irritation effects of the test item were evaluated according to the Draize method (OECD No.: 405, 2012). Rabbits were treated with analgesic and anaesthetic as per the regulatory guideline. Two animals were used to make the classification according to the Sponsor request. The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. A volume of 0.1 mL of the test item was administered as a single dose. The eyes were examined at 1, 24, 48 and 72 hours after the application. Initial Pain Reaction (IPR/PR) was not observed. First animal (No: 00166) clinical observation At one hour after the application: Conjunctival redness (score 2), chemosis (score 1) and discharge (score 3) were found. At 24 hours after the application: Conjunctival redness (score 1) was found. At 48 and 72 hours after the application There were no clinical signs, and no conjunctival or corneal effects observed. The effects were fully reversible therefore the animal was euthanized. Second animal (No: 00159) clinical observation At one hour after the application: Conjunctival redness (score 2), chemosis (score 1) and discharge (score 3) were found. At 24 hours after the application: Conjunctival redness (score 1) was found. At 48 and 72 hours after the application There were no clinical signs, and no conjunctival or corneal effects observed. The effects were fully reversible therefore the animal was euthanized. There were no clinical sign of systemic toxicity observed in the animals during the study and no mortality occurred. During the study, the control eye of each animal was symptom-free. The body weights of the animal were considered to be within the normal range of variability. The animal individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

chemosis : 0.00, 0.00

discharge : 0.00, 0.00

redness : 0.33, 0.33

cornea opacity : 0.00, 0.00

iris : 0.00, 0.00

The test item, applied to rabbit eye mucosa, caused conjunctival redness and transient discharge. The effects were fully reversible within 48 hours. The substance is not classified as irritating based on these results.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:

K1: The study was performed according to OECD guidelines and GLP.

Justification for selection of eye irritation endpoint:

K1: The study was performed according to OECD guidelines and GLP.

Effects on eye irritation: slightly irritating

Justification for classification or non-classification

An in vitro skin irritation test show no effects, therfore the substance is not classified for skin irritation or corrosion.

The test item, applied to rabbit eye mucosa, caused conjunctival redness and transient discharge. The effects were fully reversible within 48 hours. The substance is not classified as irritating based on these results.