Registration Dossier

Administrative data

Description of key information

Skin Irritation

In vitro skin irritation (OECD 439) - no indication of skin irritation.

In vivo skin irritation (OECD 404) - no indication of skin irritation.

Eye Irritation

In vitro eye irritation (equivalent to OECD 492) - no indication of irritation.

In vivo irritation (OECD 405) - no indication of irritation.

Based on the weight of evidence, the data indicates that the Test Item should not be classified as a skin irritant nor an eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 August 2013 - 15 October 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: E00031-633

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In the dark at ambient temperature
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
skin obtained from plastic surgery from multiple donors
Justification for test system used:
The EpiSkin RhE model has been accepted as a valid model for the assessment of skin irritation by European Centre for the Validation of Alternative Methods (ECVAM) and the Organisation for Economic Co-operation and Development (OECD).
Vehicle:
unchanged (no vehicle)
Details on test system:
The EpiSkin RhE tissues were produced by culturing adult human keratinocytes on a collagen base in conditions which permitted their terminal differentiation and the reconstruction of an epidermis with a functional horny layer (stratum corneum). The human keratinocytes came from mammary samples obtained from healthy consenting donors during plastic surgery. HIV 1 & 2, HEP B and HEP C tests were carried out on the donors as well as verification of the bacteriological and fungal sterility of the cells and absence of mycoplasma. This model is used to assess the irritation potential of a test item by examining the initiating events in the cascade, for example: the cytotoxicty. The test system measures the reduction of MTT to formazan metabolite by mitochondrial reductase and, therefore, irritant materials are identified by their ability to reduce cell viability below a threshold of 50% below the negative control value.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 µL

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL (Dulbecco's PBS)

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL (aqueous SDS solution (5%, w/v))
Duration of treatment / exposure:
15 minutes
Duration of post-treatment incubation (if applicable):
42.5 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 99.9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: No visible damage
- Direct-MTT reduction: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
Acceptance criteria: OD: >/=0.6 and - Acceptance criteria met for positive control: yes
Acceptance criteria: mean % viability: - Acceptance criteria met for variability between replicate measurements: yes
Interpretation of results:
GHS criteria not met
Conclusions:
EXP0700332 is non-irritant when tested in the EpiSkin in vitro irritiation assay
Executive summary:

The dermal irritation potential was assessed by applying EXP0700332 (10 µL) to the exposed surface of three EpiSkin reconstructed human epidermis (RhE) units for 15 min. The surface area of the EpiSkin was 0.38 cm^2, therefore the application rate was 26.3 µL/cm^2. After the 15 min exposure period, the surface of the EpiSkin was rinsed with Dulbecco’s phosphate-buffered saline (PBS; 25 mL). Initial rinsing with PBS did not remove all of the applied dose from the EpiSkin surface. Therefore, cotton swabs soaked in PBS were used to gently remove any remaining test item from the surface of the EpiSkin units. The EpiSkin was then incubated in maintenance media (2 mL) for a recovery period of 42.5 hours in a humidified incubator set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively. Following incubation, the EpiSkin units were transferred to assay medium (2 mL) containing MTT (0.3 mg/mL) and returned to the incubator (set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively) for 3 hours. Biopsies of the EpiSkin membranes were removed, added to acidified isopropanol and refrigerated for ca 68 h in order to extract the formazan. Formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control, aqueous sodium dodecyl sulphate (SDS) solution (5%, w/v), and the negative control, PBS, were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual EpiSkin tissue was calculated as a percentage of the mean negative control viability (defined as 100%). Exposure to EXP0700332 resulted in a mean EpiSkin viability of 99.90% +/- 9.42% of the negative control value. Exposure to the positive control, aqueous SDS solution (5%, w/v), resulted in a mean EpiSkin viability of 10.62% +/- 3.54% of the negative control value.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 February 2014 - 19 May 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: E00031-633
- Expiration date of the lot/batch: 31 Dec 2014
- Purity test date: 99%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient temperature in the dark
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan UK Lts, Bicester, UK
- Age at study initiation: 8 - 9 months
- Weight at study initiation: 2.699 kg - 3.468 kg
- Housing: Singly housed in appropriately sized floor pens.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 3 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 19 °C
- Humidity (%): 44 - 46%
- Air changes (per hr): 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: From: 18 Feb 2017 To: 21 Feb 2014
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
4 hour exposure
Observation period:
All sites were examined for irritation at approximately 1, 24, 48 and 72 hours after patch removal and on Day 8. Additional examination was performed on Days 14 and 15 for Animals 2 and 1 repectively.
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: The animals were clipped, approximately 8 cm x 8 cm, across the dorsal area of the trunk the day before dosing. 2 gauze patches (2.5 cm x 2.5 cm) were applied to the rabbits topically in the upper dorsal trunk region of the tested site and the lower dorsal trunk region of the control site.
- Type of wrap if used: Each patch was covered with Micropore semi-occlusive tap. An elastic banage was also wrapped around the torso of the rabbit to keep the patches in place.

REMOVAL OF TEST SUBSTANCE
The patches were removed after 4 hours and the test sites delineated. The test and control sites were washed with sterile water.

OBSERVATION TIME POINTS
The animals were observed daily for any clinical / behavioural signs.
All application sites were examined at 1, 24, 48, 72 hours after patch removal and on Day 8. Additional examination look place for Animals 1 and 2 on Days 15 and 14 respectively.

SCORING SYSTEM:
- Method of calculation: Dermal Scoring System
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 1.3
Max. score:
2
Reversibility:
fully reversible within: 8 days
Remarks:
Dry, flaking skin was observed on Days 7 and 8
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 1
Max. score:
1
Reversibility:
fully reversible within: 8 days
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 1
Max. score:
1
Reversibility:
fully reversible within: 8 days
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
other: Not Applicable as no edema seen
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
other: Not Applicable as no edema seen
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
other: Not Applicable as no edema seen
Remarks on result:
no indication of irritation
Interpretation of results:
GHS criteria not met
Conclusions:
Applying the GHS and ECHA criteria for classification of skin irritants, EXP0700332 was not categorised as irritating to skin.
Executive summary:

This study investigated the skin irritation and/or corrosive potential of the test item, EXP0700332, after a single application to New Zealand White rabbits. Three female rabbits were subjected to a 4 h exposure of 0.5 mL of the test item, EXP0700332. The test item was applied, as supplied, onto dorsal trunk under a semiocclusive patch. To act as a control, another area of the trunk was similarly treated, except that no test item was applied. All sites were examined for irritation at approximately 1, 24, 48 and 72 h after patch removal. Examinations continued for 2 animals up to 14 days (Day 15) after patch removal in order to evaluate the reversibility of reactions.

Erythema was present at the application site of all rabbits 24 h, 48 h and 72 h after patch removal. The degree of severity was very slight (Grade 1) erythema, with the exception of one animal at the 48 h observation, when well defined (Grade 2) erythema was recorded. No erythema was seen at the application site of any rabbit after the 72 h observation timepoint, and there was no oedema recorded in any rabbit at any observation timepoint.

Using the criteria described in the Regulation (EC) 1272/2008 under the conditions of the study, EXP0700332 would be considered to be non-irritating to rabbit skin. Therefore, no symbol and no risk phrase are required. In addition, applying the GHS and ECHA criteria for classification of skin irritants, EXP0700332 was not categorised as irritating to skin.

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

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 August 2013 to 07 October 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
Deviations:
yes
Remarks:
Study performed before the guideline came into force.
Principles of method if other than guideline:
Evaluation of ocular irritation is part of the Human Health Hazard Assessment required for registration of a chemical. In this study, the irritation potential of EXP0700332 was assessed using the HCE® in vitro ocular irritation test.

The SkinEthic HCE® model assesses the irritation potential of a test item by examining the cytotoxic effects of the test item after a defined exposure period and recovery time. The endpoint of the HCE® assay is the estimation of cell viability by assaying the reduction of MTT by mitochondrial enzymes. Irritant materials are identified by their ability to reduce cell viability below a threshold of 50% of the negative control value.

30 µL of EXP0700332, the postitive control and the negative control were applied directly onto the surface of HCE tissue using a positive displacement pipette.The tissue was exposed for 60 minutes before rinsing with PBS (25 mL). After washing, all HSE tissues were transferred to fresh maintenance medium and incubated for 18 hours at 5% CO2 and 37 degrees Celsius. The tissues were then transferred to a new 24 well plate containing MTT solution in SkinEthic maintenance medium (0.5 mg/mL, 300 µL per well). The HCE® tissues were then incubated for 3 hours in a humidified incubator set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively. The cells were then removed from the MTT solution and gently dried before being transferred to wells containing 750 µL of isopropanol to extract the formazan for 90 minutes. The absorbance was then measured to determine the cell viability.
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: E00031-633
- Retest date of the lot/batch: 30 January 2014

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient temperature in the dark
Species:
human
Strain:
other: Human Corneal Epithelium
Details on test animals or tissues and environmental conditions:
Evaluation of ocular irritation is part of the Human Health Hazard Assessment required for registration of a chemical. In this study, the irritation potential of EXP0700332 was assessed using the HCE® in vitro ocular irritation test.
The SkinEthic reconstructed Human Corneal Epithelium (HCE) model has undergone ECVAM prevalidation testing and has been shown to have a high correlation with existing in vivo and in vitro data for known eye irritants and non-irritant compounds.

The SkinEthic HCE® model assesses the irritation potential of a test item by examining the cytotoxic effects of the test item after a defined exposure period and recovery time. The endpoint of the HCE® assay is the estimation of cell viability by assaying the reduction of MTT by mitochondrial enzymes. Irritant materials are identified by their ability to reduce cell viability below a threshold of 50% of the negative control value.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
30 µL (application rate of 60 µL/cm2)
Duration of treatment / exposure:
60 minutes
Observation period (in vivo):
Not Applicable
Duration of post- treatment incubation (in vitro):
After exposure of the test substance, the HCE was washed with 25 mL PBS and incubated for 18 hours at 37 degrees celsius at 5% CO2.
Number of animals or in vitro replicates:
3 replicates per treatment
Details on study design:
EXP0700332, the positive control (Triton X-100 solution; 20%, w/v) and negative control (PBS), were each applied directly (30 µL) onto the surface of four HCE tissues using a positive displacement pipette. The pipette tip was used to gently spread the applied doses over the entire exposed area of the HCE tissues. The surface area of the EpiSkin was 0.5 cm2. Therefore, the mean application rate was 60 µL/cm2 for all treatments.

After exposure to the test item and controls for 60 minutes, the exposure period was terminated by rinsing the HCE® tissues with PBS (25 mL). After washing, all HCE® tissues were transferred to fresh maintenance medium and incubated for 18 hours in a humidified incubator set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively.

After the recovery period, the HCE® tissues were transferred to a new 24 well plate containing MTT solution in SkinEthic maintenance medium (0.5 mg/mL, 300 µL per well). The HCE® tissues were then incubated for 3 hours in a humidified incubator set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively. After the incubation, each HCE® tissue was removed from the MTT solution and gently tapped dry to remove any excess moisture. The HCE® tissues were then transferred to wells containing 750 µL isopropanol to extract the formazan. A further 750 µL of isopropanol was also added to the upper surface of each HCE® tissue. The wells were then sealed and left for 90 minutes at room temperature, protected from light, with gentle shaking. Following the solvent extraction two aliquots were added to a 96 well plate for each sample. Plates were analysed using an MRX plate reader using wavelength measurement at 550 nm. Absorbance values were calculated against the background isopropanol sample contained on the plate.
Irritation parameter:
other: Cell viability (%)
Value:
ca. 100.81
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
Interpretation of results:
GHS criteria not met
Conclusions:
EXP0700332 was demonstrated to be non-irritant to eyes when tested in vitro using the HCE® reconstructed human corneal model.
Executive summary:

Evaluation of ocular irritation is part of the Human Health Hazard Assessment required for registration of a chemical. In this study, the irritation potential of EXP0700332 was assessed using the HCE® in vitro ocular irritation test.

Prior to the conduct of the irritation assay, a preliminary test was conducted to assess the intrinsic ability of the test item to reduce methylthiazoldiphenyl-tetrazolium bromide (MTT) to formazan, which is a measure of cell viability in the assay. The results of the MTT direct reduction test showed that EXP0700332 did not reduce MTT to formazan.

Eye irritation potential was assessed by applying EXP0700332 (30 µL) to the exposed surface of three HCE tissues for 60 minutes. The surface area of the HCE was 0.5 cm2, therefore the application rate was 60 µL/cm^2. After the 60 minutes exposure period, the test item was washed from the surface of the HCE using Dulbecco’s phosphate-buffered saline (PBS; 25 mL) and cotton swabs. The HCE tissues were then incubated for a recovery period of 16 hours in a humidified incubator set to maintain temperature and CO2 levels of 37 degrees Celsius and 5%, respectively. Following incubation, the HCE tissues were transferred to maintenance medium containing MTT (0.5 mg/mL) and returned to the incubator for 3 hours. The HCE tissues were then transferred to isopropanol in order to extract the formazan. After extraction (90 minutes), the formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control,

Triton X-100 solution (20%, w/v), and the negative control, PBS, were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual HCE tissue was calculated as a percentage of the mean negative control viability (defined as 100%).

Exposure to EXP0700332 resulted in a mean HCE® viability of 100.81 +/- 7.23% of the negative control value. Exposure to the positive control, Triton X-100 (20%, w/v), resulted in a mean HCE® viability of 0.23 +/- 0.11% of the negative control value. Cell viability values below a threshold of 50% of the negative control viability indicate that the test item is irritant.

In conclusion, EXP0700332 was demonstrated to be non-irritant to eyes when tested in vitro using the HCE® reconstructed human corneal model.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 October 2013 to 07 January 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- lot/batch No.of test material: E00031-633
- Retest date of the lot/batch: 30 January 2014
- Purity test date: 99%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At ambient temperature in the dark
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan UK Ltd, Bicester, UK.
- Age at study initiation:15 to 24 weeks old
- Weight at study initiation: 2.572 kg and 4.158 kg
- Housing: Singly housed in appropriately sized stainless steel cages with a 'Noryl' dual level interior and perforated floor
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approximately 20°C
- Humidity (%): approximately 44% to 60%
- Air changes (per hr): minimum of 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: From: 22 October 2013 To: 25 October 2013
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
72 hours
Observation period (in vivo):
72 hours (observed at 1, 4, 24, 48 and 72 hour time points after instillation)
Duration of post- treatment incubation (in vitro):
Not applicable
Number of animals or in vitro replicates:
2 animals
Details on study design:
Both eyes of the rabbits were examined the day before dosing (Day -1) and no ocular defects were detected. In order to provide a therapeutic level of systemic analgesia, both animals received a subcutaneous injection of an analgesic (0.01 mg/kg buprenorphine) between 66 min and 78 min before test item instillation. Between 6 and 20 min before instillation, a topical ocular anaesthetic, 0.5% tetracaine hydrochloride, was dropped into each eye.

EXP0700332 was instilled into the right eye; the test item was placed into the eye by gently pulling the lower eyelid away from the eyeball to form a sac into which the test item was dropped. The lids were then gently held together for 1 to 2 s in order to prevent loss of the test item. The left eye remained untreated toserve as a control.
Approximately 8 hours after instillation, both animals received a subcutaneous injection of 0.5 mg/kg meloxicam in order to provide a continued therapeutic level of systemic analgesia. After examination by a veterinary surgeon, it was decided that, because neither animal showed any clinical signs of pain and distress, it was not necessary to administer buprenorphine at 8 h post-instillation. Since neither animal showed any positive ocular lesions or clinical signs of pain and distress at the 24 h post instillation assessment, no further analgesics were administered.

An Ocular Scoring system was used to grade the ocular reactions.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible within: 24 hours
Remarks on result:
no indication of irritation
Remarks:
Redness grade 1 was seen at observation time period 1 hour and 4 hours but was fully reversbile within 24 hours.
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible within: 24 hours
Remarks on result:
no indication of irritation
Remarks:
Redness grade 1 was seen at 1 and 4 hour observation time periods which fully reversed within 24 hours
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
other: Discharge
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible within: 24 hours
Remarks on result:
no indication of irritation
Remarks:
Garde 1 discharge seen at observation time points 1 and 4 hours but reversed within 24 hours
Irritation parameter:
other: Discharge
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Max. score:
0
Reversibility:
fully reversible within: 24 hours
Remarks on result:
no indication of irritation
Remarks:
Grade 1 chemosis was oberseved at 1 and 4 time observation periods which fully reversed within 24 hours
Interpretation of results:
GHS criteria not met
Conclusions:
Using the criteria described in the European Union Council Directive 67/548/EEC, under the conditions of the study, EXP0700332 would be considered to be non-irritating to the rabbit eye. Therefore, no symbol and no risk phrase are required. In addition, applying the GHS and ECHA criteria for classification of eye irritants,
EXP0700332 was not categorised as irritating to eyes.
Executive summary:

This study investigated the eye irritation potential of EXP0700332 after a single instillation to the rabbit eye.

Two male rabbits were treated with 0.1 mL of EXP0700332, instilled into the conjunctival sac of the right eye. The left eye remained untreated and hence it acted as a control. Both eyes were examined for evidence of irritation approximately 1, 4, 24, 48 and 72 h after instillation.

No irritation was noted in the control eyes of either rabbit at any observation timepoint. There were no reactions in the cornea or iris of the treated eye in either animal. Slight conjunctival redness, accompanied by slight discharge, was seen in the treated eye of

both rabbits at 1 and 4 h after instillation. No other signs of eye irritation were recorded in either rabbit at any other observation timepoint.

Using the criteria described in the Regulation (EC) 1272/2008, under the conditions of the study, EXP0700332 would be considered to be non-irritating to the rabbit eye. Therefore, no symbol and no risk phrase are required. In addition, applying the GHS and ECHA criteria for classification of eye irritants, EXP0700332 was not categorised as irritating to eyes.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

The in vitro skin irritation study conducted in accordance with OECD guideline 439, using the EpiSkin test system showed the registration substance to be non irritating.

The in vivo skin irritation study was conducted in accordance with OECD guideline 404, using New Zealand White rabbits. Grade 1 erythema was present but was fully reversible within 8 days and no oedema was recorded at any observation timepoint. EXP0700332 would be considered as non-irritating using the critera described in the Regulation (EC)1272/2008.

Eye irritation

The in vitro eye irritation study was conducted in equivalence with the OECD 492 study as this test was performed before the guideline came into force. No irritation was observed.

The in vitro eye irritation study was conducted in accordance with the OECD 405 guideline and to GLP standards. Grade 1 redness and discharge was seen in both animals but fully reversed within 24 hours. Under the critera described in the Regulation (EC) 1272/2008, EXP0700332 would be considered as non-irritating.

Justification for classification or non-classification

Neither the in vitro or in vivo tests produced an signs of dermal irritation that would warrant classification under Regulation (EC) No 1272/2008.

Neither the in vitro or in vivo tests produced signs or eye irritation that would warrant classification under Regulation (EC) No 1272/2008.