Registration Dossier

Administrative data

Description of key information

Skin irritation/corrosion: not irritating (OECD 439, GLP, K, rel. 1).

Eye irritation: not irritating, WoE approach (OECD 437 + OECD 405 on a supporting substance, both rel.1).

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:
From 2012-10-02 to 2012-10-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Monitoring Programme (Inspected on 2012-07-10)
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
Following the REACH bottom-up strategy, the EPISKIN™ Reconstructed Human Epidermis Model method was used to assess skin irritation as recommended in the OECD test guideline No. 439.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis Model Kit
- Tissue batch number(s): 12-EKIN-036
- Production date: not reported
- Shipping date: 02 October 2012
- Delivery date: 02 October 2012
- Date of initiation of testing: 02 October 2012

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: not reported
At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test item.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL in DPBS
- Incubation time: 3 hours
- Spectrophotometer: Anthos 2001 microplate reader
- Wavelength: 540 nm (without a reference filter)
- Filter: not applicable
- Filter bandwidth: not applicable
- Linear OD range of spectrophotometer: no data reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: negative control OD values: Negative Control Item 0.630, 0.612, 0.691 (historical (124 studies) mean of the negative control was 0.797 ± 0.086; The lowest value was 0.603 and the highest was 1.315).
- Barrier function: IC50= 2.1 mg/mL ( ≥ 1.5 mg/mL)
- Morphology: well differenciated epidermis consisting of a basal layer, several spinous and granular layers and a thick stratum corneum
- Contamination: : absence of bacteria, fungus and mycoplasma
- Reproducibility: not reported

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE: the test item did not directly reduce MTT

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritant to skin if the viability after the 15-min exposure period followed by the 42h post-exposure period is less or equal to 50%
- The test substance is considered to be non-irritant to skin if the viability after the 15-min exposure period followed by the 42h post-exposure period is greater than 50%
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
- Amount applied: 10±2 mg of the solid test item.
Formulations will be used within two hours of preparation and will be assumed to be stable for this period. The concentration, stability and homogeneity of the formulations will not be determined by analysis.
Duration of treatment / exposure:
15 ± 0.5 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3 epidermis/product
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minute exposure period and 42 h post-exposure incubation period
Value:
64.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
6.8%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: not reported

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD540 for the negative control treated tissues was 0.644 and the standard deviation value of the percentage viability was 6.4%. The negative control acceptance criterion was therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 6.8% relative to the negative control treated tissues and the standard deviation value of the percentage viability was 1.2%. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: The standard deviation calculated from individual percentage tissue viabilities of the three identically treated tissues was 7.6%. The test item acceptance criterion was therefore satisfied.
- Historical control data:
Since October 2009 to the commencement of this study (124 studies) the historical mean OD540 of the positive control was 0.059 ± 0.022 and the mean viability was 7.4 ± 3.2%. In this same period the historical mean OD540 of the negative control was 0.797 ± 0.086.
Also in the same historical period the lowest OD540 of the positive control was 0.025 and the highest 0.175 and the lowest percentage viability was 1.3 and the highest was 15.6. The lowest OD540 of the negative control was 0.603 and the highest was 1.315.
In this study the mean OD540 for the positive control was 0.044 and the mean viability was 6.8%. The mean OD540 of the negative control was 0.644.
The positive and negative controls were therefore within the historical ranges for this testing facility.

Table 7.3.1/1: Mean OD540 Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item

Item

OD540 of tissues

Mean OD540 of triplicate tissues

±SD of OD540

Relative individual tissue viability (%)

Relative mean viability (%)

± SD of Relative mean viability (%)

Negative Control Item

0.630

0.644

0.041

97.8

100*

6.4

0.612

95.0

0.691

107.3

Positive Control Item

0.039

0.044

0.008

6.1

6.8

1.2

0.053

8.2

0.039

6.1

Test Item

0.372

0.416

0.049

57.8

64.7

7.6

0.408

63.4

0.469

72.8

SD=   Standard deviation

*=     The mean viability of the negative control tissues is set at 100%

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the test substance is not classified for skin irritation according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

The purpose of this test was to evaluate the skin irritation potential of ST 10 C 08 using the EPISKINTMreconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. The principle of the assay is based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT (3‑[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt to a blue formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls.

This test was designed to be compatible with the Method B.46 of Commission Regulation (EC) No. 440/2008/EC and was performed in compliance with GLP.

Triplicate tissues were treated with ST 10 C 08 for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 hours. At the end of the post‑exposure incubation period each tissue was taken for MTT-loading. After MTT loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT‑loaded tissues. 

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µl samples were transferred to the appropriate wells of a pre‑labelled 96‑well plate. The optical density was measured at 540 nm.

The relative mean viability of the test item treated tissues was 64.7 ± 7.6 %, after the 15‑minute exposure period.

The quality criteria required for acceptance of results in the test were satisfied.

Under the experimental conditions of this study, the test substance is not classified for skin irritation according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

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:
weight of evidence
Study period:
2012-11-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Monitoring Programme (inspected on 2012-07-10)
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF BOVINE EYES
- Source: from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter and placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 µg/mL).
- Age: adult cattle, typically between 12 and 60 months

STORAGE AND TRANSPORT CONDITIONS
The eyes were transported to the test facility over ice packs and used on the same day as slaughter. Transfer of the bovine eyes from the abattoir to the test facility is less than 45 minutes. Decomposition of the corneas is not expected to occur during this transfer period.
Vehicle:
physiological saline
Controls:
yes, concurrent vehicle
yes, concurrent positive control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): 20%

VEHICLE
- Concentration (if solution): 0.9% w/v
Duration of treatment / exposure:
240 minutes
Observation period (in vivo):
NA
Duration of post- treatment incubation (in vitro):
NA
Number of animals or in vitro replicates:
3 cornea per treatment groups
Details on study design:
SELECTION AND PREPARATION OF CORNEAS
- Examination: macroscopically, only corneas free of damage were used.
- Preparation of the eyes: as described in the guideline

QUALITY CHECK OF THE ISOLATED CORNEAS
- Quality (opacitometer): Corneas that displayed an initial opacity reading greater than 7 units were discarded.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: yes

POSITIVE CONTROL USED: yes

APPLICATION DOSE AND EXPOSURE TIME
For the purpose of this study the test item was prepared as a 20% dilution in 0.9% w/v sodium chloride solution. The MEM was removed from the anterior chamber of the BCOP holder and the test item preparation or control items were applied to the cornea. The holders were gently tilted back and forth to ensure a uniform application of the item over the entire cornea. Each holder was incubated, anterior chamber uppermost, at 32 ± 1ºC for 240 minutes.

TREATMENT METHOD: closed chamber method

POST-INCUBATION PERIOD: no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: At the end of the exposure period the test item preparation and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM.
- POST-EXPOSURE INCUBATION: NA

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The change in opacity for each cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final opacity reading. These values were then corrected by subtracting the average change in opacity observed for the negative control corneas. The mean opacity value of each treatment group was then calculated by averaging the corrected opacity values of each cornea for that treatment group.
- Corneal permeability: passage of sodium fluorescein dye
The corrected OD492 was calculated by subtracting the mean OD492 of the negative control corneas from the OD492 value of each treated cornea. The OD492 value of each treatment group was calculated by averaging the corrected OD492 values of the treated corneas for the treatment group.
- Others (e.g, pertinent visual observations, histopathology): Visual observation: The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
The following formula was used to determine the In Vitro Irritancy Score:
In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)
Additionally, the opacity and permeability values were evaluated independently to determine whether the test item induced a response through only one of the two endpoints.


DECISION CRITERIA:
A test item that induces an In Vitro Irritancy Score ≥ 55.1 is defined as an ocular corrosive or severe irritant.
For an acceptable test the following positive and negative control criterion must be achieved:
20% w/v Imidazole was used for positive control purposes. The test was acceptable if the positive control produced an In Vitro Irritancy Score which fell within two standard deviations of the historical mean of 103.4 for this testing facility. Therefore the In Vitro Irritancy Score should fall within the range of 66.1 to 140.8.
0.9% w/v sodium chloride solution was used for negative control purposes. The test was acceptable if the negative control produced an In Vitro Irritancy Score which is less than or equal to the highest In Vitro Irritancy Score for this testing facility during 2012 up to and including this study ≤6.1.
Irritation parameter:
in vitro irritation score
Run / experiment:
mean/3corneas
Value:
8.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
IVIS = 1.9
Positive controls validity:
valid
Remarks:
IVIS = 86.3
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: the test item did not cause any opacity or permeability of the corneas compared with the results of the negative control

DEMONSTRATION OF TECHNICAL PROFICIENCY: not included in the report

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values if different from the ones specified in the test guideline: not included in the report

Table 7.3.2/1: Individual and Mean Corneal Opacity and Permeability Measurements

Treatment

Cornea Number

Opacity

Permeability (OD)

In vitro Irritancy Score

Pre-Treatment

Post-Treatment

Post-Treatment-Pre‑Treatment

Corrected Value

 

Corrected Value

Negative Control

1

3

5

2

 

0.052

 

 

2

2

3

1

 

0.034

 

 

3

3

4

1

 

0.037

 

 

 

 

 

1.3*

 

0.041¨

 

1.9

Positive Control

4

1

68

67

65.7

0.950

0.909

 

5

1

69

68

66.7

1.750

1.709

 

6

1

64

63

61.7

1.755

1.714

 

 

 

 

 

64.7·

 

1.444·

86.3

Test Item

7

4

7

3

1.7

0.396

0.355

 

8

2

8

6

4.7

0.531

0.490

 

9

1

7

6

4.7

0.197

0.156

 

 

 

 

 

3.7·

 

0.334·

8.7


OD= Optical density

* = Mean of the post treatment - pre‑treatment values

° = Mean permeability

#= Mean corrected value

Interpretation of results:
GHS criteria not met
Conclusions:
ST 10 C 08 was considered not to be an ocular corrosive or severe irritant.
Executive summary:

A study was performed to assess the ocular irritancy potential of the test item to the isolated bovine cornea. The study was conducted according to the OECD guideline No. 437 and in compliance with GLP.

ST 10 C 08 was applied at ac concentration of 20% w/v for 240 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

The in vitro Irritancy scores are summarised as follows:

Treatment

In Vitro Irritancy Score

Test Item

8.7

Negative Control

1.9

Positive Control

86.3

 With an IVIS < 55.1, the test item was considered not to be an ocular corrosive or severe irritant.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 1989-02-10 to 1989-02-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study performed according to OECD test guideline No. 405 and in compliance with GLP.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, Fuellinsdorf, Switzerland
- Age at study initiation: M: 15 weeks; F: 16 weeks
- Weight at study initiation: M: 2.7 kg; F: 2.6-2.9 kg
- Housing: individually in stainless steel cages
- Diet (e.g. ad libitum): Pelleted standard Kliba 341, Batch 42/88 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): community tap water from Itingen ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 +/- 3 °C
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From:1989-02-10 To: 1989-02-21
Vehicle:
other: undiluted or in Neantine
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Left eye: 0.1 g (undiluted) / Right eye: 0.1 mL (diluted into 30 % in Neantine)
Duration of treatment / exposure:
Eyes were not rinsed
Observation period (in vivo):
Eyes were observed 1, 24, 48, 72 hours and 7 days after administration
Number of animals or in vitro replicates:
1 male and 2 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done
- Time after start of exposure:

SCORING SYSTEM: Draize scale according to the OECD test guideline No. 405

TOOL USED TO ASSESS SCORE: slit-lamp and diagnostic-lamp
Irritation parameter:
cornea opacity score
Basis:
animal: #1, #2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: #1, #2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: #1 and #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 h
Remarks on result:
probability of weak irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
chemosis score
Basis:
animal: #1 and #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
probability of weak irritation
Irritant / corrosive response data:
See Table 7.3.2/1
Other effects:
none

Table 7.3.2/1: Mean irritant/corrosive response data of each animal at each observation time up to removal from the test

 

Score at time point / Reversibility

Cornea opacity(/4)

Iris

(/2)

Conjunctivae

Redness

(/3)

Chemosis

(/4)

1 h

0/0/0

0/0/0

2/2/2

1/0/1

24 h

0/0/0

0/0/0

1/1/2

0/0/1

48 h

0/0/0

0/0/0

0/0/2

0/0/1

72 h

0/0/0

0/0/0

0/0/1

0/0/0

Mean 24-48-72 hrs

0.0/0.0/0.0

0.0/0.0/0.0

0.33/0.33/1.67

0.00/0.00/0.67

Reversibility*)

-

-

c.

c.

Average time (unit) for reversion

 

 

7 days

72 hours

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test material is not classified as irritating to eyes according to the criteria of the Regulation (EC) No 1272/2008 (CLP) and to the GHS.
Executive summary:

In an eye irritation study performed according to the OECD guideline No. 405, and in compliance with GLP, 0.1 g of undiluted Fixambrene was instilled into the left eye of 3 New Zealand White Rabbit. The eyes were not rinsed after the instillation of the test item.The left eye of each rabbit was instilled with 0.1 mL Fixambrene diluted at 30 % in Neantine. Animals were observed at 1, 24, 48 and 72 hours after dosing under a standard light source. The reactions in the conjunctiva (redness, chemosis and discharge), the iris and the cornea (opacity and area involved) were scored according to the Draize scale. 

 

The calculated mean score for each individual lesion for each animal treated with undiluted Fixambrene within 3 scoring times (24, 48 and 72 h) were as follows: 0.0/0.0/0.67 for chemosis; 0.33/0.33/1.67 for redness; 0.0/0.0/0.0 for iris and corneal lesions.

The effects observed were all reversible within 7 days.

 

Under the test conditions, Fixambrene is not classified as irritating to eyes according to the criteria of the Regulation (EC) No 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

Endpoint:
eye irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
[Further information is included as attachment to Iuclid section 13]

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across approach is based on the hypothesis that the source and target substances have similar physico-chemical, toxicological, ecotoxicological and environmental fate properties because of
their structural similarity.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target and the source substances (Table 1) are Cycloalkane ethers. The target substance is the racemic form, while the source substance is a reaction mass of the source together with the 9bS isomer.

3. ANALOGUE APPROACH JUSTIFICATION
The source substance is expected to have similar toxicological profile than the target substance considering the respective physico-chemical and toxicological properties. The study design (OECD 405, GLP) is adequate and reliable for the purpose of the prediction based on read-across. The test material used represents the source substance as described in the hypothesis in terms of purity and impurities. The results of the studies are adequate for the purpose of classification and labelling.
Therefore, based on the considerations above, it can be concluded that the results of the acute dermal toxicity study conducted in the rat with the source substance are likely to predict the properties of the target substance and are considered as adequate to fulfil the information requirement of Annex VIII, 8.2

4. DATA MATRIX
See Iuclid section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Remarks:
Read-across justification document
Irritation parameter:
cornea opacity score
Basis:
animal: #1, #2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: #1, #2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: #1 and #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 h
Remarks on result:
probability of weak irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
chemosis score
Basis:
animal: #1 and #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
probability of weak irritation
Irritant / corrosive response data:
See Table 7.3.2/1
Other effects:
none

Table 7.3.2/1: Mean irritant/corrosive response data of each animal at each observation time up to removal from the test

 

Score at time point / Reversibility

Cornea opacity(/4)

Iris

(/2)

Conjunctivae

Redness

(/3)

Chemosis

(/4)

1 h

0/0/0

0/0/0

2/2/2

1/0/1

24 h

0/0/0

0/0/0

1/1/2

0/0/1

48 h

0/0/0

0/0/0

0/0/2

0/0/1

72 h

0/0/0

0/0/0

0/0/1

0/0/0

Mean 24-48-72 hrs

0.0/0.0/0.0

0.0/0.0/0.0

0.33/0.33/1.67

0.00/0.00/0.67

Reversibility*)

-

-

c.

c.

Average time (unit) for reversion

 

 

7 days

72 hours

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the source substance is not classified as irritating to eyes according to the criteria of the Regulation (EC) No 1272/2008 (CLP) and to the GHS.
Executive summary:

In an eye irritation study performed according to the OECD guideline No. 405, and in compliance with GLP, 0.1 g of undiluted source substance was instilled into the left eye of 3 New Zealand White Rabbit. The eyes were not rinsed after the instillation of the test item.The left eye of each rabbit was instilled with 0.1 mL Fixambrene diluted at 30 % in Neantine. Animals were observed at 1, 24, 48 and 72 hours after dosing under a standard light source. The reactions in the conjunctiva (redness, chemosis and discharge), the iris and the cornea (opacity and area involved) were scored according to the Draize scale. 

 

The calculated mean score for each individual lesion for each animal treated with undiluted Fixambrene within 3 scoring times (24, 48 and 72 h) were as follows: 0.0/0.0/0.67 for chemosis; 0.33/0.33/1.67 for redness; 0.0/0.0/0.0 for iris and corneal lesions.

The effects observed were all reversible within 7 days.

 

Under the test conditions, the source subtance is not classified as irritating to eyes according to the criteria of the Regulation (EC) No 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

A key study was identified (Harlan, 2012). The purpose of this test was to evaluate the skin irritation potential of ST 10 C 08 using the EPISKIN reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours.

This test was designed to be compatible with the Method B.46 of Commission Regulation (EC) No. 440/2008/EC and was performed in compliance with GLP. The quality criteria required for acceptance of results in the test were satisfied. The relative mean viability of the test item treated tissues was 64.7 ± 7.6 %, after the 15‑minute exposure period. With a tissue viability > 50%, ST 10 C 08 was considered to be non-irritant to skin.

Eye irritation:

The Integrated Testing Strategy as described in ECHA guidance chapter R.7A was followed to assess the eye irritation potential of the substance. In the absence of any alert based on physico-chemical properties, existing human data, existing animal studies and structurally-related substances, a new in vitro test was performed (Harlan, 2013, Rel.1). This BCOP assay was conducted according to the OECD guideline No. 437 and in compliance with GLP. Under the conditions of the test, with an In Vitro Irritancy Score < 55.1, ST 10 C 08 was considered not to be an ocular corrosive or severe irritant. On the basis of this result, a read-across to the in vivo study conducted on a supporting substance (mixture of isomers, including approximately 50% of the registered substance) was used to avoid unnecessary animal testing (see §Toxicokinetics for read-across justification).

In this eye irritation study (RCC, 1989, Rel. 1) performed according to the OECD guideline No. 405, and in compliance with GLP, 0.1 g of undiluted Fixambrene was instilled into the eye of 3 rabbits. The eyes were not rinsed after the instillation of the test item. The calculated mean score for each individual lesion for each animal treated with undiluted Fixambrene within 3 scoring times (24, 48 and 72 h) were as follows: 0.0/0.0/0.67 for chemosis; 0.33/0.33/1.67 for redness; 0.0/0.0/0.0 for iris and corneal lesions. The effects observed were all reversible within 7 days. Under the test conditions, Fixambrene is not classified as irritating to eyes.

Using a weight of evidence approach, the substance is therefore not classified for eye irritation.

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 (CLP).

Self classification:

Based on the available data no additional self-classification is proposed regarding both skin and eye irritation according to the CLP and the GHS.

No data was available regarding respiratory irritation, however the substance not being classified for skin and eye irritation, no classification is expected for respiratory irritation.