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Description of key information

Skin irritation: Clevios K Primer W8 dry is irritant to skin according to UN GHS and EU CLP regulation.

Skin corrosion: Clevios K Primer W8 dry is non corrosive to skin according to EU CLP and UN GHS.

Eye irriatation: Clevios K Primer W8 dry is classified as serious eye damaging (EU CLP/UN GHS Category 1).

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
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
Updated Guideline adopted 29 July 2016
Qualifier:
according to guideline
Guideline:
other: MatTek test protocol "In vitro EpiDerm™ Skin Corrosion Test (EPI-200-SCT)", 07 November 2014
Deviations:
yes
Remarks:
According to the quality certificate of the supplier of the test kit demonstrating its robustness, during treatment with 1% Triton X-100 the ET50 was determined as 4.77 hours ≤ ET50 ≤ 8.72 hours instead of 4.0 hours ≤ ET50 ≤ 8.7 hours.
GLP compliance:
yes (incl. QA statement)
Test system:
human skin model
Source species:
human
Details on animal used as source of test system:
The epidermis model (e.g. EpiDermTM) is derived from human keratinocytes and consists of normal, human-derived epidermal keratinocytes (NHEK) which have been cultured to form a multilayered, highly differentiated model of the human epidermis. The NHEK, which are cultured on specially prepared cell culture inserts using serum free medium, attain levels of differentiation at the cutting edge of in vitro skin technology. Ultrastructurally, the skin models closely parallel human skin.
Justification for test system used:
Validation studies have shown that tests employing human skin models are able to discriminate between known skin corrosives (Optional sub-category 1A, optional sub-category 1B and 1C) and non-corrosives. The test protocol may also enable the distinction between severe and less severe skin corrosives.
Species:
other: not applicable to the Human Epidermis Test Method
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
after 3 minutes exposure
Value:
> 50
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
after 1 hour exposure
Value:
> 15
Other effects / acceptance of results:
The optical pre-experiment (colour interference pre-experiment) to investigate the test item’s colour change potential in water did not led to a change in colour.

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour.

The test item is considered to be non-corrosive to skin:
• since the viability after 3 minutes exposure is greater than 50% and
• the viability after 1 hour exposure is greater than 15%.

The acceptance criteria are met:
• the mean OD of the tissue replicates treated with the negative control is ≥ 0.8 and ≤ 2.8 for every exposure time (values between 1.409 and 1.649)
• the mean viability of the tissue replicates treated with the positive control for 1 hour, is <15% compared to the negative control (2.3%)
• the Coefficient of Variation (CV) in the range 20 – 100% viability between tissue replicates is ≤ 30% (range: 0.1% to 11.4%)
Interpretation of results:
GHS criteria not met
Conclusions:
According to the results of this study and under the reported experimental conditions, Clevios K Primer W8 dry is non corrosive to skin according to EU CLP and UN GHS.
Executive summary:

The in vitro study was performed to assess the corrosive potential of Clevios K Primer W8 dry by means of the Human Skin Model Test with EpiDerm™ tissues models.

The test item passed the MTT- and the colour interference pre-tests.

Independent duplicate tissues of EpiDermTM were exposed to the test item, the negative control (deionised water) or the positive control (8.0 N KOH) for 3 minutes and 1 hour, respectively.

Afterwards, the test and the control items were rinsed off the tissues, and a 3 hour incubation period (37 ± 1 °C, 5 ± 0.5 % CO2) with MTT solution followed. MTT solution was then aspirated from the wells and the wells were rinsed with DPBS. Inserts were transferred into new 24 well plates. The formazan salt was extracted 2 hours at room temperature.

The required acceptability criteria were met.

Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period (25.8%) and for the 1 hour exposure period (2.3%) thus confirming the validity of the test system and the specific batch of tissue models.

After exposure to the test item Clevios K Primer W8 dry the relative absorbance value decreased to 94.4% after 3 minutes exposure. After 1 hour exposure the relative absorbance value was reduced to 61.4%. Both values did not fall below the threshold for corrosivity which is defined to be 50% after the 3 minutes exposure and 15% after the 1 hour exposure. Therefore, the test item is not considered to be corrosive.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Identification: Clevios K Primer W8 dry
Batch: 9002784195
CAS Name: 1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)
CAS No.: 1326716-46-4
Purity: 99%
Physical State / Appearance: White, solid
Expiry Date: 01 March 2020
Storage Conditions: At room temperature
Stability in solvent Stable in water and ethanol for approx. 24 hours at room temperature
Purpose of Use: Industrial chemical
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Treatment of Epi-200-SIT Component with Clevios K Primer W8 dry
Value:
27.24
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Treatment of Epi-200-SIT Components with DPBS (negative control)
Value:
100
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Treatment of Epi-200-SIT Components with 5% SDS solution in deionized water (positive control)
Value:
4.17
Other effects / acceptance of results:
This in vitro study was performed to assess the irritation potential of Clevios K Primer W8 dry by means of the Human Skin Model Test.

The test item passed the MTT- and the Colour Interference pre-tests.

The test item, the negative control (DPBS), and the positive control (5% SDS) were applied to triplicate tissue, respectively.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 42 hours, the tissues were treated with the MTT solution for 3 hours followed by 2 hours of extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control, the absorbance values were well within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance compared with the negative control to 4.17% thus ensuring the validity of the test system.

The standard deviations between the three tissue percentage viability values of each group (test item, positive and negative controls) in the main test were below 18 (threshold according OECD 439: ≤ 18), thus ensuring the validity of the study.

Compared to the negative control the mean relative viability was reduced to 27.24% after exposure of the skin tissues to the test item. This value is below the threshold for irritancy of ≤ 50%. Therefore, the test item is considered to possess an irritant potential.
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
It can be stated that based on this OECD 439 study and under the experimental conditions reported, Clevios K Primer W8 dry is irritant to skin.
Executive summary:

This in vitro study was performed to assess the irritation potential of Clevios K Primer W8 dry by means of the Human Skin Model Test.

The test item did not reduce MTT (pre-test for direct MTT reduction), and it did not change colour when mixed with deionised water (pre-test for colour interference). Also, its intrinsic colour was not intensive. Consequently, additional tests with freeze-killed or viable tissues were not necessary.

Each three tissues of the human skin model EpiDerm™ were treated with the test item, the negative control (Dulbecco's Phosphate Buffered Systeme (DPBS)) or the positive control (5% Sodium Dodecyl Sulphate (SDS)) for 60 minutes.

After treatment with the negative control the absorbance values were well within the required range of the acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval, thus assuring the quality of the tissues.

Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the 60 minutes treatment interval, thus assuring the validity of the test system.

After treatment with the test item Clevios K Primer W8 dry the mean relative viability value decreased to 27.24% compared to the relative absorbance value of the negative control. This value is below the threshold for irritancy of ≤ 50%. Therefore, the test item is considered to possess an irritant potential.

In conclusion, it can be stated that in this study and under the experimental conditions reported, Clevios K Primer W8 is irritant to 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
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)
Deviations:
yes
Remarks:
The positive control is 10% (w/v) Benzalkonium chloride (purity not indicated by the producer) in saline since the laboratory historical control data is established with this chemical.
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
GLP compliance:
yes (incl. QA statement)
Irritation parameter:
in vitro irritation score
Value:
284.56
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
The calculated mean in vitro irritancy score was 284.56. According to OECD 437 (see table in chapter 3.8.3) the test item is classified as serious eye damaging (EU CLP/UN GHS Category 1).
Executive summary:

This in vitro study was performed to assess the corneal irritation and damage potential of Clevios K Primer W8 dry by means of the BCOP assay using fresh bovine corneae.

After a first opacity measurement of the fresh bovine corneae (t0), the 20% (w/v) suspension in saline (0.9% (w/v) NaCl in deionised water) of the test item Clevios K Primer W8 dry as well as the positive and the negative controls were each applied to different corneae fixed in an incubation chamber in horizontal position and incubated for 240 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t240).

After the opacity measurements, permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.

With the negative control (physiological saline), neither an increase of opacity nor permeability of the corneae was observed.

The positive control (10% (w/v) benzalkonium chloride in saline) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damage (EU CLP/UN GHS Category 1).

Relative to the negative control, the test item Clevios K Primer W8 dry caused an increase of the corneal opacity and permeability. The calculated mean in vitro irritancy score was 284.56. According to OECD 437 (see table in chapter 3.8.3) the test item is classified as serious eye damaging (EU CLP/UN GHS Category 1).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Justification for classification or non-classification

Skin irritation:

According to the results of the In vitro Skin Irritation Test according to OECD guideline 439 and under the reported experimental conditions, Clevios K Primer W8 dry is irritant to skin according to UN GHS and EU CLP regulation.

Skin corrosion:

According to the results of the In vitro Skin Corrosion Test according to guideline OECD 431 and under the reported experimental conditions, Clevios K Primer W8 dry is non corrosive to skin according to EU CLP and UN GHS.

Eye irriatation:

Relative to the negative control, the test item Clevios K Primer W8 dry caused an increase of the corneal opacity and permeability. The calculated mean in vitro irritancy score was 284.56. According to OECD 437 (see table in chapter 3.8.3) the test item is classified as serious eye damaging (EU CLP/UN GHS Category 1).