Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Non skin irritating

Not eye 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
Study period:
From June 12 to July 02, 2003
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
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted April 24, 2002
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne / France.
- Age at study initiation: 22-23 weeks old male; 9-10 and 20-21 weeks old females.
- Housing: individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4646 were provided for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet, ad libitum (batch no.28103) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water: community tap water from FÜllinsdorf, ad libitum.
- Acclimation period: under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study. Four days before treatment, one flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was again clipped; animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 *C
- Humidity: 30-70 %
- Air changes: approximately 10-15 air changes per hour.
- Photoperiod: the animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark.
- Other: music was played during the daytime light period.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.5 g (per animal) of test item was weighed as delivered by the sponsor and then moistened with approximately 0.1 ml of purified water before application.
The pH of the test item was measured before the study initiation date. A formulation of 1 % in water was prepared. The pH was found to be 5 to 6.
Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
1 male and 2 females
Details on study design:
TEST SITE
- Area of exposure: 0.5 g of test item was placed on a surgical gauze patch (ca. 4 cm x 4 cm). The gauze patch was applied to the intact skin of the clipped area.
- Type of wrap if used: the patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing: the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours

OBSERVATIONS
- Mortality: daily from acclimatization of the animals to the termination of test.
- Clinical signs: daily from acclimatization of the animals to the termination of test.
- Body weights: at start of acclimatization, on the day of application and at termination of observation.

OBSERVATION TIME POINTS
The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after the removal of the dressing, gauze patch and test item.

SCORING SYSTEM
Grading of Skin Reactions
ERYTHEMA AND ESCHAR FORMATION
No erythema 0
Very slight erythema 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) or eschar formation (injuries in depth preventing erythema) reading 4

OEDEMA FORMATION
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (edges raised approximately 1 mm) 3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure) 4
Irritation parameter:
erythema score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2.3
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2.3
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
With the exception of the yellow staining, the test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

COLORATION
Slight yellow staining produced by the test item of the treated skin was observed in all animal from the 1-hour reading up to 10 days after treatment. Slight yellow staining was still present in two animals at the 14 day examination, the end of the observation period.

CORROSION
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.
Other effects:
VIABILITY/MORTALITY/CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

BODY WEIGHTS
The body weights of all rabbits were considered to be within the normal range of variability. One animal (no. 69) slightly lost body weight (1.4 %) between the treatment start and the end of the study. The body weilnt loss was considered to be incidental and treatment unrelated.

Individual reactions

Animal Reaction After
1 hr 24 hrs 48 hrs 72 hrs 7 days 10 days 14 days
67 M Erythema 0 0 0 0 0 0 0
68 F Erythema 0 0 0 0 0 0 0
69 F Erythema 0 0 0 0 0 0 0
67 M Oedema 0 0 0 0 0 0 0
68 F Oedema 0 0 0 0 0 0 0
69 F Oedema 0 0 0 0 0 0 0
Interpretation of results:
other: not classified, according to the CLP Regulation (EC) No 1272/2008
Conclusions:
Not irritating
Executive summary:

The primary skin irritation potential of the substance was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The durition of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after removal of the dressing.

With the exception of the yellow staining the test item did not elicit any skin reactions at the application sitä of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

The test item caused yellow staining of the treated skin in all animals. This effect was not reversible and was still evident in two animals 14 days after treatment, the end of the observation period for all animals. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no other clinical signs of test item related effects were observed.

Conclusion

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 06, 2001), with the exception of the yellow staining which persisted in two animals throughout the observation period, test item is considered to be "not irritating" to rabbit skin.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From September 12th to October 17th, 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
2018
GLP compliance:
yes (incl. QA statement)
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
0.03 g
Duration of treatment / exposure:
10 s then frinse with ca. 20 ml isotonic saline
Duration of post- treatment incubation (in vitro):
up to 240 minutes after post-treatment rinse.
Details on study design:
SELECTION AND PREPARATION OF ISOLATED EYES
Eyes selection
After removing the head from the plastic box, it was put on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. One small drop of fluorescein solution 2 (w/v) % was applied onto the cornea surface for a few seconds and subsequently rinsed off with 20 ml isotonic saline. Then the fluorescein-treated cornea was examined with hand-held slit lamp or slit lamp microscope, with the eye in the head, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit.

Preparation of eyes
The eyeball was carefully removed from the orbit by holding the nictitating membrane with surgical forceps, while cutting the eye muscles with bent scissors without cutting off the optical nerve too short. The procedure avoided pressure on the eye in order to prevent distortion of the cornea and subsequent corneal opacity. Once removed from the orbit, the eye was placed onto damp paper and the nictitating membrane was cut away with other connective tissue. The prepared eyes were kept on the wet papers in a closed box so that the appropriate humidity was maintained.

Eyes examination and acclimatization time
The prepared eye was placed in a steel clamp with the cornea positioned vertically with the eye in the correct relative position (same position as in the chicken head). Again avoiding too much pressure on the eye by the clamp. Because of the relatively firm sclera of the chicken eyeball, only slight pressure was applied to fix the eye properly. The clamp with the eyeball was transferred to a chamber of the superfusion apparatus. The clamp holding the eye was positioned in such a way that the entire cornea was supplied with saline solution dripping from a stainless steel tube, at a rate of approximately 3 to 4 drops/minutes. The door of the chamber was closed except for manipulations and examinations, to maintain temperature and humidity.
The appropriate numbers of eyes were selected and after being placed in the superfusion apparatus, the selected eyes were examined again with the slit lamp microscope to ensure that they were in good condition. The focus was adjusted to see clearly the isotonic saline which was flowing on the cornea surface. Eyes with a high baseline fluorescein staining (i.e., > 0.5) or corneal opacity score (i.e., > 0.5) were rejected. The cornea thickness was measured using the depth measuring device on the slit lamp microscope (Haag-Streit BQ 900) with the slit-width set at 9½, equalling 0.095 mm. Any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced. If the selected eyes were appropriate for the test, acclimatization started and was conducted for approximately 45 to 60 minutes. The temperature was verified to be in the range of 32 ± 1.5 °C in all chambers during the acclimatization and treatment periods.


EQUILIBRATION AND BASELINE RECORDINGS
At the end of the acclimatization period, a zero reference measurement was recorded for cornea thickness and opacity to serve as a baseline (t=0) for each individual eye. The cornea thickness of the eyes should not change by more than ±5-7 % within approximately 45 to 60 minutes before the start of application. Slight changes in thickness (0% to 2%) were observed in the eyes during the first and additional experiment, finding considered as normal when maintaining enucleated eyes. Following the equilibration period, the fluorescein retention was measured. Baseline values were required to evaluate any potential test item related effects after treatment. The location of any minor findings was marked on the record sheet as a drawing, if applicable. None of the eyes were discarded as no eye was considered unsuitable after the baseline assessment.

NUMBER OF REPLICATES: 3 eyes per test item, 3 eyes per positive control, 1 negative control eye

NEGATIVE CONTROL USED : yes

POSITIVE CONTROL USED : yes, imidazole

APPLICATION DOSE AND EXPOSURE TIME
After the zero reference measurements in each experiment, one out of three test item treated eyes was taken out of its chamber and placed on a layer of tissue with the cornea facing upwards. The eye was held in horizontal position over a container to catch waste, while the test item was applied onto the centre of the cornea. The test item was applied in an amount of 0.03 g by attempting to cover the cornea surface uniformly with the test substance, while taking care not to damage or touch the cornea with the application equipment. This procedure was repeated for each test item treated eye.
The three positive control eyes were treated in a similar way with 0.03 g imidazole.
One negative control eye was treated with saline solution. The saline solution was applied in a volume of 30 µl from micropipette, in such a way that the entire surface of the cornea was covered with negative control, taking care not to damage or touch the cornea with the application equipment.

OBSERVATION PERIOD
The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within ±5 minutes were considered acceptable.
The cornea thickness and cornea opacity were measured at all time points. Fluorescein retention was determined at baseline (t=0) and 30 minutes after the post-treatment rinse.

REMOVAL OF TEST SUBSTANCE
The time of application was monitored, then after an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with 20 ml saline solution at ambient temperature, while taking care not to damage the cornea but attempting to remove all the residual test item if possible. The eye in the holder was then returned to its chamber. The time while the eye was out of the chamber was limited to the minimum.

The imidazole and test item were stuck on the corneas’ surface in all eyes at 30 minutes after the post-treatment rinse. The gentle rinsing with 20 ml saline was performed in all imidazole and test item treated eyes after the 30, 75, 120 and 180 minutes of observation. The imidazole and test item treated cornea surfaces were not totally cleared at 240 minutes after the post-treatment rinse in the first and additional experiment.


Evaluation
The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects. Results from corneal opacity, swelling, and fluorescein retention were evaluated separately to generate an ICE (Isolated Chicken Eye) class for each endpoint. The ICE classes for each endpoint were then combined to generate an Irritancy Classification for each test substance.

Cornea swelling was calculated according to the following formulae

CS at time t = 100 × (CT at time t – CT at t=0) CT at t=0
Mean CS at time t = (FECS(at time t)+ SECS (at time t) + TECS (at time t)) / 3

Remark:
CS = Cornea swelling
CT = Cornea thickness
FECS = First eye cornea swelling
SECS = Second eye cornea swelling
TECS = Third eye cornea swelling
Mean CS = The mean percentage of corneal swelling
at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse
at t=0 = Reference value

For the calculation of Maximum Swelling, small negative numbers for swelling (0 to -5%) following application are counted as zero. Large negative numbers (> -12 %) are probably due to erosion and indicate a severe effect (scored as class IV). Cases of values of -5 % to -12 % are evaluated on a case by case basis but in the absence of other findings do not indicate a severe effect.

Cornea opacity was calculated according to the following formulae
Scores were taken at any given timepoint according to:
Score Observation
0 No opacity
0.5 Very faint opacity
1 Scattered or diffuse areas; details of the iris are clearly visible
2 Easily discernible translucent area; details of the iris are slightly obscured
3 Severe corneal opacity; no specific details of the iris are visible; size of the pupil is barely discernible
4 Complete corneal opacity; iris invisible

ΔCO at time t = CO at time t - CO at t=0
Mean CO(at time t) = (FEΔCO (at time t)+ SEΔCO(at time t) + TEΔCO(at time t)) / 3

Remark:
CO = Cornea opacity
ΔCO = Difference between cornea opacity and cornea opacity reference value
FEΔCO = Difference between first eye cornea opacity and first eye cornea opacity reference value
SEΔCO = Difference between second eye cornea opacity and second eye cornea opacity reference value
TEΔCO = Difference between third eye cornea opacity and third eye cornea opacity reference value
Mean CO = The mean corneal opacity value
at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse
at t=0 = Reference value

Fluorescein retention was calculated according to the following formulae
Scores were taken at any given timepoint according to:
Score Observation
0 No fluorescein retention
0.5 Very minor single cell staining
1 Single cell staining scattered throughout the treated area of the cornea
2 Focal or confluent dense single cell staining
3 Confluent large areas of the cornea retaining fluorescein

ΔFR at time t = FR at time t – FR at t=0
Mean FR = (FEΔFR (at time t) + SEΔFR(at time t) + TEΔFR at time t)) / 3

Remark:
FR = Fluorescein retention
ΔFR = Difference between fluorescein retention and fluorescein retention reference value
FEΔFR = Difference between first eye fluorescein retention and first eye fluorescein retention reference value
SEΔFR = Difference between second eye fluorescein retention and second eye fluorescein retention reference value
TEΔFR = Difference between third eye fluorescein retention and third eye fluorescein retention reference value
Mean FR = The mean fluorescein retention value
at time t = Observation time at 30 minutes after the post-treatment rinse
at t=0 = Reference value
Irritation parameter:
cornea opacity score
Run / experiment:
I
Value:
0.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Remarks:
up to 240 min
Run / experiment:
I
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
I
Value:
0
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
II
Value:
1.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Remarks:
up to 240 min
Run / experiment:
II
Value:
3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
II
Value:
0.3
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:
The mean values of the treated eyes for maximum corneal thickness change, corneal opacity, fluorescein retention and other observation (morphological effect etc.) are given below.

First experiment:
Test item: Fluorescent Brightener 367
Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 3% I
Mean maximum corneal swelling at up to 240 min 4% I
Mean maximum corneal opacity 0.7 II
Mean fluorescein retention 0.0 I
Other observations none
Overall ICE Class 2×I, 1×II

Positive control: imidazole
Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 33% IV
Mean maximum corneal swelling at up to 240 min 40% IV
Mean maximum corneal opacity 4.0 IV
Mean fluorescein retention 3.0 IV
Other observations Corneal opacity score 4 was observed in three eyes eye at 30 minutes after the post-treatment rinse.
Overall ICE Class 3×IV

The positive control solution was classed as corrosive/severely irritating, UN GHS Classification: Category 1.

Negative control: NaCl (9 g/l saline)
Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 0% I
Mean maximum corneal swelling at up to 240 min 0% I
Mean maximum corneal opacity 0.0 I
Mean fluorescein retention 0.0 I
Other observations None
Overall ICE Class 3×I

The negative control NaCl (9 g/l saline) had no significant effects on the chicken eye in this study.

Additional experiment:
Test Item: Fluorescent Brightener 367

Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 2% I
Mean maximum corneal swelling at up to 240 min 3% I
Mean maximum corneal opacity 1.2 II
Mean fluorescein retention 0.3 I
Other observations None
Overall ICE Class 2×I, 1×II


Positive control: imidazole
Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 40% IV
Mean maximum corneal swelling at up to 240 min 44% IV
Mean maximum corneal opacity 4.0 IV
Mean fluorescein retention 3.0 IV
Other observations Corneal opacity score 4 was observed in three eyes at 30 minutes after the post-treatment rinse.
Overall ICE Class 3×IV

The positive control solution was classed as corrosive/severely irritating, UN GHS Classification: Category 1.

Negative control: NaCl (9 g/l saline)
Observation Value ICE Class
Mean maximum corneal swelling at up to 75 min 0% I
Mean maximum corneal swelling at up to 240 min 0% I
Mean maximum corneal opacity 0.5 I
Mean fluorescein retention 0.0 I
Other Observations None
Overall ICE Class 3×I

The negative control NaCl (9g/l saline) had no significant effects on the chicken eye in this study.

Positive and negative control values were within the corresponding historical control data ranges.

ICE classification criteria for corneal swelling

Mean Corneal Swelling (%) ICE Class
0 to 5 I
>5 to 12 II
>12 to 18 ( >75 min after treatment ) II
>12 to 18 (<75 min after treatment ) III
>18 to 26 III
>26 to 32 ( >75 min after treatment ) III
>26 to 32 (<75 min after treatment ) IV
>32 IV

ICE classification for opacity

Mean Maximum Opacity Score ICE Class
0.0 – 0.5 I
0.6 – 1.5 II
1.6 – 2.5 III
2.6 – 4.0 IV

ICE classification criteria for mean fluorescein retention

Mean Fluorescein Retention Score at 30 minutes post - treatment ICE Class
0.0 – 0.5 I
0.6 – 1.5 II
1.6 – 2.5 III
2.6 – 3.0 IV

Overall in vitro classifications

UN GHS Classification Combinations of the 3 endpoints
No Category 3×I,
2×I, 1×II
1×I, 2×II
No prediction can be made Other combinations
Category 1(Causes serious eye damage) 3×IV
2×IV, 1×III
2×IV, 1×II
2×IV, 1×I
Corneal opacity ≥ 3 at 30 min (in at least 2 eyes)
Corneal opacity = 4 at any time point (in at least 2 eyes)
Severe loosening of the epithelium (in at least 1 eye)
Interpretation of results:
other: not eye irritant according to the CLP Regulation (EC 1272/2008)
Conclusions:
In this in vitro eye irritation study, using the Isolated Chicken Eye model with test item, no ocular corrosion or severe irritation potential was observed. The overall ICE class was 2×I, 1×II in the first and the additional experiment.

According to the guideline OECD 438, test item is not eye irritant.


Executive summary:

The isolated chicken eye test (ICET) was run according to OECD guideline 438.

Test compound was applied in a single dose (30 mg/eye) onto the cornea of isolated chicken eyes.

Tested corneas were evaluated pre-treatment and at approximately 30, 75, 120, 180, and 240 minutes after the post-treatment rinse. The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects. All of the endpoints, with the exception of fluorescein retention (which was determined only at pre-treatment and 30 minutes after test substance exposure) were determined at each time points.

According to the first experiment results the test item showed negative outcome. Based on the OECD 438 additional experiment was necessary to confirm or discard the negative outcome.

The imidazole (positive control) was ground before use in the study. The test item and positive control applied in an amount of 0.03 g/eye by powdering the entire surface of the cornea attempting to cover the cornea surface uniformly with the test substance or positive control. Three test item treated eyes, three positive control treated eyes and one negative control treated eye which was treated with 30 µl saline solution were used in the first and additional experiment.

After an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with ~20 ml saline solution at ambient temperature and this procedure was repeated for each eye.

The test item and imidazole were stuck on the corneas’ surface in all eyes at 30 minutes after the post-treatment rinse in the first and additional experiment. The imidazole treated cornea surfaces were not totally cleared at 240 minutes after the post-treatment rinse in the first and additional experiment.

In this ICET, test itemdid not cause ocular corrosion or severe irritation in the enucleated chicken eyes in either cases. The overall ICE class was 2xI, 1xII in the first and the additional experiment.

Positive and negative controls showed the expected results. The experiment was considered to be valid.

No ocular corrosion or severe irritation potential was observed. The overall ICE class was 2×I, 1×II in the first and the additional experiment.

According to the guideline OECD 438, test item does not require classification within the CLP Regulation (EC 1272/2008).

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

SKIN IRRITATION

The primary skin irritation potential of the Fluorescent Brightener 367 was investigated according to OECD guideline 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact skin of three New Zealand White rabbits. The durition of treatment was four hours.

With the exception of the yellow staining the test item did not elicit any skin reactions at the application sitä of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0. The test item caused yellow staining of the treated skin in all animals. This effect was not reversible and was still evident in two animals 14 days after treatment, the end of the observation period for all animals. However, no corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no other clinical signs of test item related effects were observed.

EYE IRRITATION

The eye irritation potential of Fluorescent Brightener 367 was evaluated according to OECD guideline 438.

The test compound was applied in a single dose (30 mg/eye) onto the cornea of isolated chicken eyes.

Tested corneas were evaluated pre-treatment and at approximately 30, 75, 120, 180, and 240 minutes after the post-treatment rinse. The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects.  

Upon negative results in the first experiment, an additional experiment was necessary to confirm or discard the negative outcome, based on OECD 438.

Imidazole was used as positive control.

The test item and positive control applied in an amount of 0.03 g/eye by powdering the entire surface of the cornea attempting to cover the cornea surface uniformly with the test substance or positive control. Three test item treated eyes, three positive control treated eyes and one negative control treated eye which was treated with 30 µl saline solution were used in the first and additional experiment.

After an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with ~20 ml saline solution at ambient temperature and this procedure was repeated for each eye.

The test item and Imidazole were stuck on the corneas’ surface in all eyes at 30 minutes after the post-treatment rinse in the first and additional experiment. The imidazole treated cornea surfaces were not totally cleared at 240 minutes after the post-treatment rinse in the first and additional experiment. Positive and negative controls were valid.

In this ICET, Fluorescent Brightener 367 did not cause ocular corrosion or severe irritation in the enucleated chicken eyes in either cases.

The overall ICE class was 2xI, 1xII in the first and the additional experiment.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

In the available study, the mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals, for both erythema/eschar and oedema reactions.

According to the CLP Regulation (EC 1272/2008), serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application.

The eye irritation potential was tested and test results were interpreted as described in OECD guideline 438. In the available ICET, no ocular corrosion or severe irritation potential was observed and the overall ICE class was 2×I and 1×II in the first and additional experiment.

In conclusion, the substance does not meet the criteria to be classified for skin and eye irritation, according to the CLP Regulation (EC 1272/2008).

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