1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

Updated Guideline adopted 29 July 2016

Test guideline 2

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 material

Test material information

Constituent 1

Reference substance name:

1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

EC Number:

855-905-0

Cas Number:

1326716-46-4

Molecular formula:

C24H40N2O6S2 (C10H24N2 • 2 C7H8O3S)

IUPAC Name:

1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

Test material form:

solid

In vitro test system

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.

Test animals

Species:

other: not applicable to the Human Epidermis Test Method

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

after 3 minutes exposure

Value:

> 50

Results 2

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%)

Applicant's summary and conclusion

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.