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Description of key information

Skin irritation:

In Vitro Skin Irritation Test With SILVIAL using A Human Skin Model dated on 2016 and performed accoridng to the OECD guideline No. 439 shows that the SILVIAL is irritant to skin in the test conditions. In vitro Skin Corrosion Test with SILVIAL using a Human Skin Modeldated on 2017 and performed accoridng to the OECD guideline No. 431 shows that the SILVIAL is not corrosive to skin in the test conditions.

Eye Irritation:

Evaluation of the eye Hazard potential of SILVIAL using the bovine Corneal Opacity and Permeability test (BCOP) dated on 2016 and perfomed according to the OECD Guideline No. 437, has determined that the SILVIAL IVIS is < 3, no classification is required for eye irritation or serious eye damage

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 May 2016 to 06 June 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Principles of method if other than guideline:
Any deviations from standard operating procedures were evaluated and filed in the study file. There were no deviations from standard operating procedures that affected the integrity of the study.
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Name (as stated in the report): Silvial
Batch No.: SC00017272
Expiration date: 23 October 2016
Test system:
human skin model
Remarks:
EPISKIN Small Model TM
Source species:
human
Cell type:
other: adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen.
Cell source:
other: EPISKIN Small ModelTM (EPISKIN-SMTM, 0.38 cm2, Batch no.: 16-EKIN-022)
Source strain:
other: SkinEthic Laboratories, Lyon, France
Justification for test system used:
In the interest of sound science and animal welfare, a sequential testing strategy is recommended to
minimise the need of in vivo testing. One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD and EC).
Vehicle:
unchanged (no vehicle)
Details on test system:
EPISKIN Small ModelTM (EPISKIN-SMTM, 0.38 cm2, Batch no.: 16-EKIN-022)
This model is a three-dimensional human epidermis model, which consists of adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type
I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
The liquid test item was applied undiluted (25 μl) directly on top of the tissue.
Duration of treatment / exposure:
15 ± 0.5 minutes
Duration of post-treatment incubation (if applicable):
42 hours at 37°C.
Number of replicates:
The test was performed on a total of 3 tissues per test item together with negative and positive controls.
Irritation / corrosion parameter:
% tissue viability
Remarks:
Mean tissue viability (percentage of control)
Value:
26
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
Since the mean relative tissue viability for SILVIAL was below 50% it is considered to be irritant.
Other effects / acceptance of results:
The positive control had a mean cell viability after 15 ± 0.5 minutes exposure of 4.5%. The absolute mean OD570 of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Finally, it is concluded that this test is valid and that SILVIAL is irritant in the in vitro skin irritation test under the experimental conditions described in this report.
Executive summary:

In vitro skin irritation test with SILVIAL using a human skin model.

This report describes the ability of SILVIAL to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small model (EPISKIN-SMTM)). The possible skin irritation potential of SILVIAL was tested through topical application for 15 minutes.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch SC00017272 of SILVIAL was a colourless liquid. SILVIAL was applied undiluted (25 μl) directly on top of the skin tissue for 15 ± 0.5 minutes. After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

SILVIAL did interact with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT).

In addition to the normal procedure, three killed tissues treated with test item and three killed non-treated tissues were used for the cytotoxicity evaluation with MTT. The non-specific reduction of MTT by SILVIAL was -2.6% of the negative control tissues, therefore no correction was needed.

Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with SILVIAL compared to the negative control tissues was 26%. Since the mean relative tissue viability for SILVIAL was below 50% after 15 ± 0.5 minutes treatment it is considered to be irritant.

The positive control had a mean cell viability of 4.5% after 15 ± 0.5 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.

Finally, it is concluded that this test is valid and that SILVIAL is irritant in the in vitro skin irritation test under the experimental conditions described in this report.

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 Apr 2017 to 14 Apr 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Qualifier:
according to guideline
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Name (as stated in the report): Silvial
Batch No.: SC00017272
Expiration date: 13 October 2017
Test system:
human skin model
Remarks:
EpiDerm Skin Model
Source species:
human
Cell type:
other: human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.
Cell source:
other: EpiDerm Skin Model (EPI-200, Lot no.: 25761 kit U and V)
Source strain:
other: MatTek Corporation, Ashland MA, U.S.A.
Justification for test system used:
Recommended test system in international guidelines (OECD and EC).
Vehicle:
unchanged (no vehicle)
Details on test system:
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
The liquid test item was applied undiluted (50 μl) directly on top of the tissue
Duration of treatment / exposure:
Two tissues were used for a 3-minute exposure to SILVIAL and two for a 1-hour exposure.
Number of replicates:
The test was performed on a total of 4 tissues per test item together with a negative control and positive control
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
after 1-minute and 3-minute treatment
Value:
103
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: No indication of corrision
Other effects / acceptance of results:
The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance limit <=2.8) and the laboratory historical control data range . The mean relative tissue viability following the 1-hour exposure to the positive control was 7%.
In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was < 7%, indicating that the test system functioned properly

SILVIAL compared to the negative control tissues. Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after both the 3-minute and 1-hour treatments with SILVIAL compared to the negative control tissues was 103%. Because the mean relative tissue viability for SILVIAL was not below 50% after 3 minutes treatment and not below 15% after 1 hour treatment SILVIAL is considered to be not corrosive.

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, SILVIAL is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.
Executive summary:

The objective of this study was to evaluate SILVIAL for its ability to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of SILVIAL was tested through topical application for 3 minutes and 1 hour.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch SC00017272 of the test item was a colourless liquid. The test item was applied undiluted (50 μl) directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 7% after the 1-hour exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance limit <=2.8) and the laboratory historical control data range. In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was < 7%, indicating that the test system functioned properly.

Because in the skin irritation test a color change was observed by adding MTT-medium it was concluded that SILVIAL did interact with the MTT endpoint.

In addition to the normal 3-minute and 1-hour procedure, two freeze-killed tissues treated with test item and two freeze-killed negative control treated tissues were used for the cytotoxicity evaluation with MTT at each time point. The non-specific reduction of MTT by SILVIAL was 2.6% and 3.4% of the negative control tissues after 3 minutes and 1 hour respectively.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained both after 3-minute and 1-hour treatments with SILVIAL compared to the negative control tissues was 103%. Because the mean relative tissue viability for SILVIAL was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment SILVIAL is considered to be not corrosive.

In conclusion, SILVIAL is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
28 April 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Principles of method if other than guideline:
There were no deviations from standard operating procedures that affected the integrity of the study.
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Name (as stated in the report): Silvial
Batch No.: SC00017272
Expiration date: 23 October 2016
Species:
cattle
Details on test animals or tissues and environmental conditions:
Bovine eyes were used as soon as possible after slaughter. They come from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter. Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.
Vehicle:
unchanged (no vehicle)
Remarks:
The test item was tested neat.
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
Undiluted Silvial was introduced onto the epithelium of the cornea.
Duration of treatment / exposure:
The cornea were incubated in a horizontal position for 10 ± 1 minutes at 32 ± 1°C after the introduction of the test item or control.
Irritation parameter:
in vitro irritation score
Remarks:
mean in vitro irritancy score
Run / experiment:
after 10 minutes of treatement
Value:
0.3
Negative controls validity:
valid
Remarks:
ranged from 0.7 to 1.1
Positive controls validity:
valid
Remarks:
ranged from 41 to 55
Remarks on result:
no indication of irritation
Remarks:
Since SILVIAL induced an IVIS ≤ 3,
Other effects / acceptance of results:
The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 46 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.
Interpretation of results:
GHS criteria not met
Conclusions:
Since SILVIAL induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.
Executive summary:

Evaluation of the eye hazard potential of SILVIAL using the Bovine Corneal Opacity and Permeability test (BCOP test).

This report describes the potency of chemicals to induce serious eye damage using isolated bovine corneas. The eye damage of SILVIAL was tested through topical application for 10 minutes.

The study procedures described in this report were based on the most recent OECD guideline.

Batch SC00017272 of SILVIAL was a colourless liquid with a purity of 99.4%. The test item was applied as it is (750 μl) directly on top of the corneas.

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 46 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

SILVIAL did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 0.3 after 10 minutes of treatment.

Since SILVIAL induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Silvial has shown a skin irritant potential, it needs to be classified Category 2 (irritant) based on CLP classification criteria. In parallel, the Eye irritant potiential was evaluated and demontrates

SILVIAL IVIS is < 3, no classification is required for eye irritation or serious eye damage.