Registration Dossier

Administrative data

Description of key information

Skin irritation (OECD 439): not irritating
Eye irritation (OECD 405): not irritating

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:
weight of evidence
Study period:
2014-10-14 to 2014-10-20
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)
Version / remarks:
(2013)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayrisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: human-derived epidermal keratinocytes (EpiDerm)
Justification for test system used:
This test uses the EpiDerm(TM) reconstructed human epidermis model (MatTek) which consists of human keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological porperties of the upper part of the human, i.e. the epidermis.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm(TM) tissues were provided as kits (EPI-200-SIT, MatTek)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1°C
- Temperature of post-treatment incubation: 37 ± 1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: washed intensively with DPBS for 1 min
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: none

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/mL
- Incubation time: 3 h
- Spectrophotometer: plate spectrophometer
- Wavelength: 550 nm


NUMBER OF REPLICATE TISSUES: 3


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

DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 15 minutes exposure and 42 hours post-exposure is less than or equal to 50%.
- The test substance is considered to be not irritant to skin if the viability after 15 minutes exposure 42 hours post-exposure is greater than 50% and the viability.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 30 µL

NEGATIVE CONTROL
- Amount applied: 30 µL DBPS

POSITIVE CONTROL
- Amount applied: 30 µL
- Concentration: 5% (v/v)
Duration of treatment / exposure:
60 min
Duration of post-treatment incubation (if applicable):
42 h
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
test group
Value:
96.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

EXPERIMENTAL RESULTS:

 

Negative control

Positive control

Test item

Tissue sample

1

2

3

1

2

3

1

2

3

OD550

1.592

1.564

1.451

1.499

1.449

1.391

0.044

0.044

0.038

0.037

0.037

0.038

1.540

1.563

1.429

1.395

1.343

1.340

OD550(mean)

1.578

1.475

1.420

0.044

0.038

0.038

1.552

1.412

1.342

SD

0.080

0.003

0.107

OD550(mean values of replicates)

1.491

0.040

1.435

Viability (%)

100

2.7

96.2

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
In an in vitro skin irritation human skin model test (EpiDerm) according to OECD guideline 439 and in compliance with GLP, a cell viability of 96.2% was measured after 60 min exposure and 42 h post-exposure time when compared to the untreated control. Therefore, the test item should be classified as non-irritant to the skin.
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2014-09-23 to 2014-09-29
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)
Version / remarks:
(2013)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayrisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: human-derived epidermal keratinocytes (EpiSkin)
Justification for test system used:
This test uses the EPISKIN-SM(TM) reconstructed human epidermis model (SkinEthic) which consists of human keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological porperties of the upper part of the human, i.e. the epidermis.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN-SM(TM)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation: 37 ± 1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: washed with PBS
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: none

FUNCTIONAL MODEL CONDITIONS
- Barrier function: IC50 determination (SDS concentration, MTT test, n = 14): specification >= 1.5 mg/mL, result: 2.2 mg/mL
- Morphology: Histology scoring (HES stained vertical paraffin sections, n= 6): specification ≥ 19.5; result: 21.8 ± 0.3, CV= 1.3%; Well-differentiated epidermis consisting of a basal layer, several spinous and granular layers and a thick stratum corneum

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3mg/mL
- Incubation time: 3 h
- Spectrophotometer: plate spectrophometer
- Wavelength: 550 nm

NUMBER OF REPLICATE TISSUES: 3

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

DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 15 minutes exposure and 42 hours post-exposure is less than or equal to 50%.
- The test substance is considered to be not irritant to skin if the viability after 15 minutes exposure 42 hours post-exposure is greater than 50% and the viability.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 10 µL


NEGATIVE CONTROL
- Amount applied: 10 µL

POSITIVE CONTROL
- Amount applied: 10 µL
- Concentration: 5% (v/v)
Duration of treatment / exposure:
15 min
Duration of post-treatment incubation (if applicable):
42 h
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
test group
Value:
105.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

EXPERIMENTAL RESULTS:

 

Negative control

Positive control

Test item

Tissue sample

1

2

3

1

2

3

1

2

3

OD550

0.915

0.849

0.870

0.814

0.801

0.774

0.044

0.052

0.036

0.039

0.040

0.045

0.869

0.973

0.801

0.800

0.951

0.918

OD550(mean)

0.882

0.842

0.788

0.048

0.038

0.043

0.921

0.800

0.934

SD

0.047

0.005

0.074

OD550(mean values of replicates)

0.837

0.043

0.865

Viability (%)

100

5.1

105.7

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
In an in vitro skin irritation human skin model test (EpiSkin) according to OECD guideline 439 and in compliance with GLP, a cell viability of 105.7% was measured after 15 min exposure and 42 h post-exposure when compared to the untreated control. Therefore, the test item should be classified as non-irritant.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
20 May - 04 Jun 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
(2012)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
(1998)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River France (L’Arbresle Cedex, France)
- Age at study initiation: 12 – 24 weeks
- Weight at study initiation: at least 1500 g
- Housing: animals were individually housed in labeled cages with perforated floors and shelters (Ebeco, Germany)
- Diet: pelleted diet for rabbits (Global Diet 2030, Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day; hay (TecniLab-BMI BV, Someren, The Netherlands) and wooden sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study period.
- Water: (tap/filtered) water, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 – 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
single application without washing
Observation period (in vivo):
7 days
Reading time points: 1, 24, 48 and 72 h and day 7
Number of animals or in vitro replicates:
3 males
Details on study design:
SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritant / corrosive response data:
Instillation of 0.1 mL of the test substance into one eye of each of three rabbits resulted in effects on the cornea (grade 0.33) and iris (grade 0.33) in one animal and effects of the conjunctivae in all animals (grade 1.67, 1.67 and 0.67). The corneal injury consisted of opacity and epithelial damage. The corneal injury resolved within 7 days. Iridial irritation was observed at 24 hours and was resolved at 48 hours after instillation. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 72 hours in one animal and within 7 days in the other two animals. There was no evidence of ocular corrosion. No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1: Individual eye irritation scores

Animal

Time after dosing

Cornea

 (0 – 4)

Iris

 (0 – 2)

Conjunctivae

(0 – 3)

Chemosis

(0 – 4)

 

 

1

1 hour

24 hours

48 hours

72 hours

7 days

mean 24+48+72 hours

0

0

1

0

0

0.33

0

1

0

0

0

0.33

2

2

2

1

0

1.67

2

1

1

0

0

0.67

 

 

2

1 hour

24 hours

48 hours

72 hours

7 days

mean 24+48+72 hours

0

0

0

0

0

0

0

0

0

0

0

0

2

2

2

1

0

1.67

2

1

0

0

0

0.33

 

3

1 hour

24 hours

48 hours

72 hours

mean 24+48+72 hours

0

0

0

0

0

0

0

0

0

0

2

1

1

0

0.67

1

0

0

0

0

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
In an in vivo acute eye irritation/corrosion study in the rabbit according to OECD guideline 405 and in compliance with GLP no signs of systemic toxicity were observed in the animals during the test period and no mortality occurred. Therefore, the test material is considered to be classified as a non-irritant to eyes.
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2014-08-26
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)
Version / remarks:
(2013)
Deviations:
no
GLP compliance:
yes
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST METHOD
The bovine corneal opacity and permeability (BCOP) test is an in-vitro test method used for identifying i) chemicals inducing serious eye damage and ii) chemicals not requiring classification for eye irritation or serious eye damage. The potential of a test substance to cause ocular corrosivity or severe irritancy is measured by its ability to induce opacity and increased permeability in an isolated bovine cornea. The opacity and permeability assessments of the cornea are combined to derive an in-vitro irritancy score (IVIS), which is used to classify the irritancy level of the test substance.

IDENTIFICATION OF THE SOURCE OF THE EYES, STORAGE AND TRANSPORT CONDITIONS
- Source: Vitelco, 's Hertogenbosch, The Netherlands
- Transport medium and temperature conditions: physiological saline in a suitable container under cooled conditions

PREPARATION OF THE EYES (BEFORE EXPOSURE)
- Eyes free of defects (scratches, neovascularisation): yes
- Dissection of the eyes and treatment: Isolated corneas were mounted in cornea holders
- Type of cornea holder used: MC2, Clermont, France
- Description of the cornea holder: The cornea holders consist of an anterior and a posterior compartment, which interface with the epithelial and endothelial sides of the cornea
- Test medium and temperature conditions used in the cornea holder:
cMEM (Eagle’s Minimum Essential Medium) supplemented with 1% [v/v] fetal calf serum and 1% [v/v] L-glutamine; prior to use: pre-warmed to 32 ± 1 °C
- Equilibration time: 1 h at 32 ± 1 °C

DETERMINATION OF THE INITIAL OPACITY
- Method: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer
- Specification of the device: opacitometer (OP-KIT, MC2, Clermont, France)
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount applied in the test: 750 µL
- Concentration: liquids: 100%

VEHICLE
- Substance: physiol. saline
- Concentration: 0.9% NaCl solution in deionised water
- Amount applied in the test: 750 µL

POSITIVE SUBSTANCE
- Substance: Benzalkonium Chloride
- Concentration: 10% (w/v) Benzalkonium Chloride in physiol. saline
- Amount applied in the test: 750 µL
Duration of treatment / exposure:
10 min
Observation period (in vivo):
Not applicable
Duration of post- treatment incubation (in vitro):
120 min
Number of animals or in vitro replicates:
3
Details on study design:
SELECTION AND PREPARATION OF CORNEAS : The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light.
Those exhibiting defects were discarded. The isolated corneas were stored in a petri dish with cMEM (Eagle’s Minimum Essential Medium (Invitrogen Corporation, Breda, The Netherlands) containing 1% (v/v) L-glutamine (Invitrogen Corporation) and 1% (v/v) Foetal Bovine Serum (Invitrogen Corporation)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of MC2 (Clermont, France) with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was positioned on top of the cornea and tightened with screws. The compartments of the corneal holder were filled with cMEM of 32±1°C. The corneas were incubated for the minimum of 1 hour at 32±1°C. After the incubation period, the medium was removed from both compartments and replaced with
fresh cMEM.

QUALITY CHECK OF THE ISOLATED CORNEAS : Opacity determinations were performed on each of the corneas using an opacitometer (OP-KIT, MC2, Clermont, France). The opacity of each cornea was read against an air filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used. Three corneas were selected at random for each treatment group.

NUMBER OF REPLICATES : 3

NEGATIVE CONTROL USED : physiological saline

POSITIVE CONTROL USED : 10% (w/v) Benzalkonium Chloride in physiol. saline

APPLICATION DOSE AND EXPOSURE TIME : The medium from the anterior compartment was removed and 750 µL of either the negative control, positive control or test substance was introduced onto the epithelium of the cornea. Corneas were incubated in a horizontal position for 10±1 minutes at 32±1°C.

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: yes, 120 min

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation) until no colour change of the medium was observed and thereafter with cMEM. Possible pH effects of the test substance on the corneas were recorded.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The opacitometer determined the difference in the light transmission between each control or treated cornea and an air filled chamber. The numerical opacity value (arbitrary unit) was displayed and recorded. The change in opacity for each individual cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final post-treatment reading. The corrected opacity for each positive control or test substance treated cornea was calculated by subtracting the average change in opacity of the negative control corneas from the change in opacity of each positive control or test substance treated cornea. The mean opacity value of each treatment group was calculated by averaging the corrected opacity values of the treated corneas for each treatment group.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader at OD490

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: decision criteria as indicated in the TG was used
Irritation parameter:
in vitro irritation score
Run / experiment:
mean of 3 measures
Value:
9.6
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes


The corneas treated with the test substance showed opacity values ranging from 7 to 11 and permeability values ranging from 0.025 to 0.048. The corneas were translucent after the 10 minutes of treatment with the test substance. A pH effect of the test substance was observed on the rinsing medium, the corneas were rinsed until no colour change of the medium was observed. Hence, the in vitro irritancy scores ranged from 7.7 to 11.4 after 10 minutes of treatment with the test substance.

SUMMARY OF OPACITY, PERMEABILITY AND IN VITRO IRRITATION SCORE

Treatment

Mean

Opacity1

Mean

Permeability1

Mean In vitro Irritation Score1, 2

Negative control

-1

0.000

-1.0

Positive control

(Benzalkonium Chloride)

77

3.168

124.5

Test substance

9

0.040

9.6

1: Calculated using the negative control mean opacity and mean permeability values. 2: In vitroirritancy score (IVIS) = mean opacity value + (15 x mean OD490value).

OPACITY VALUES:

Parameter

Initial opacity

Final opacity

Opacity change

Mean opacity value

Negative control

1

2

1

-1

3

0

-3

1

1

0

Test substance

1

9

8

9

0

7

7

0

11

11

Positive control

1

80

79

77

0

78

78

-1

74

75

PERMEABILITY SCORE INDIVIDUAL VALUES (CORRECTED)

Eye

Dilution factor

Negative control corrected OD49011

Negative control corrected OD49021

Negative control corrected OD49031

Negative control corrected OD490

Average

Negative control corrected final

OD490

Average OD

Negative control

1

1

0.003

0.006

0.003

0.004

0.004

0.000

2

1

-0.010

-0.010

-0.005

-0.008

-0.008

3

1

0.004

0.002

0.006

0.004

0.004

Positive control(Benzalkonium Chloride)

4

6

0.607

0.571

0.613

0.597

3.582

3.168

5

6

0.469

0.461

0.469

0.466

2.796

6

6

0.534

0.517

0.513

0.521

3.126

Methyl [3-(trimethoxysilyl)propyl]carbamate

13

1

0.052

0.042

0.049

0.048

0.048

0.040

14

1

0.046

0.046

0.050

0.047

0.047

15

1

0.024

0.026

0.025

0.025

0.025

1     OD490values corrected for the mean final negative control permeability (0.004).

IN VITRO IRRITANCY SCORE:

 

IVIS

Mean IVIS

Test substance

8.7

9.27

7.7

11.4

Positive control

132.7

124.83

119.9

121.9

HISTORICAL CONTROL DATA FOR THE BCOP STUDIES

Negative control

Positive control

Opacity

Permeability

In vitro Irritancy Score

In vitro Irritancy Score

Range

-3 – 2

-0.020 – 0.021

-3.1 – 2.3

81 – 264

Mean

-0.14

0.00

-0.14

133

SD

0.85

0.01

0.91

23

n

103

92

97

98

SD = Standard deviation

n = Number of observations

Interpretation of results:
study cannot be used for classification
Conclusions:
In an in vitro bovine corneal opacity and permeability test (BCOP test) according to OECD guideline 437 and in compliance with GLP in vitro irritancy scores ranging from 7.7 to 11.4 after 10 minutes of treatment with the test material were observed. Since the test material induced an IVIS > 3 ≤ 55, no prediction on the classification can be made. Therefore, it is concluded that an in vivo study is required for classification.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion

Two in vitro skin irritation studies with methyl-N-[3-(trimethoxysilyl)propyl]carbamate (CAS 23432-62-4) are available.

In the in vitro skin irritation study performed according to OECD 439 and in compliance with GLP human-derived epidermal keratinocytes (EpiSkin) were exposed to 10 µl of the neat test material for 15 minutes followed by a post-treatment incubation period of 42 hours (BSL Bioservice, 2015a). The irritation potential of the test material was predicted from the relative mean cell viabilities compared to the mean viability of the negative control. Positive (5% SDS) and negative (PBS) controls were included in the study and gave the expected results. The relative mean cell viability for the test material was calculated to be 105.7% when compared to the negative control after an incubation period of 42 hours. Therefore, the test material is considered to be non-irritating under the conditions of the test.

In the in vitro key skin irritation study performed according to OECD 439 and in compliance with GLP human-derived epidermal keratinocytes (EpiDerm) were exposed to 30 µl of the neat test material for 60 minutes followed by a post-treatment incubation period of 42 hours (BSL Bioservice, 2015b). The irritation potential of the test material was predicted from the relative mean cell viabilities compared to the mean viability of the negative control. Positive (5% SDS) and negative (DPBS) controls were included in the study and gave the expected results.The relative mean cell viability for the test material was calculated to be 96.2% when compared to the negative control after an incubation period of 42 hours. Therefore, the test material is considered to be non-irritating under the conditions of the test.

Based on the above study results and according to EU classification criteria, the test substance is considered to be not-irritating to skin.

 

Eye irritation

Two eye irritation studies with methyl-N-[3-(trimethoxysilyl)propyl]carbamate (CAS 23432-62-4) are available.

The eye irritation properties of methyl-N-[3-(trimethoxysilyl)propyl]carbamate (CAS 23432-62-4) have been investigated in an in vitro study performed according to OECD 437 and in compliance with GLP using the bovine corneal opacity and permeability test (BCOP, WIL, 2014a). For the assessment of the eye irritation properties 750 µl of the test material was applied to the corneal surface of bovine eyes for 10 min. Positive (10% benzalkonium chloride) and negative (0.9% NaCl) controls were included in the study and gave the expected results. Ten minutes after application mean values of cornea opacity (9), permeability (0.04) and In Vitro Irritancy Score (9.6) were calculated from three independent experiments. Hence, methyl-N-[3-(trimethoxysilyl)propyl]carbamate induced an IVIS > 3 ≤ 55 no prediction on the classification can be made. Thus, further in vivo data have been obtained.

In the in vivo key eye irritation study performed according to OECD 405 and in compliance with GLP 0.1 ml of the test material (CAS 23432-62-4) was instilled in the eye of three male New Zealand White rabbits (WIL, 2015a). The eyes were examined and the changes were graded according to the Draize scoring system 1, 24, 48 and 72 hours and 7 days post-application. Treatment with the test material resulted in effects on the cornea (grade 0.33, mean of 24, 48 and 72 h) and iris (grade 0.33, mean of 24, 48 and 72 h

) in 1/3 animals and effects on the conjunctivae in 3/3 animals (grade 1.67, 1.67 and 0.67, mean of 24, 48 and 72 h). Chemosis was recorded in 2/3 animals (grade 0.67 and 0.33, mean of 24, 48 and 72 h). The corneal injury consisted of opacity and epithelial damage, but was fully reversible within 72 hours. Iridial irritation appeared 24 hours post-application but was fully reversible within 48 hours. Conjunctivae irritation resolved within 72 hours in 1/3 and 7 days in 2/3 animals. Chemosis was fully reversible 48 and 72 hours post-application, respectively. No evidence of ocular corrosion could be detected. Moreover, no signs of systemic toxicity were observed in the animals and no mortality occurred during the course of the study.

Based on the above in vivo study results and according to EU classification criteria, the test substance is considered to be not-irritating to eyes.

Justification for classification or non-classification

Reliable data on skin and eye irritation indicate that methyl-N-[3 (trimethoxysilyl)propyl]carbamate does not meet the criteria for classification according to Regulation (EC) 1272/2008, and the available data are therefore conclusive but not sufficient for classification.