Hexamethylene diisocyanate, oligomers, reaction products with 3-methoxypropylamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From January 16, 2017 to January 20, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EU) No. 1152/2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hexamethylene diisocyanate, oligomers, reaction products with 3-methoxypropylamine

EC Number:

500-740-9

EC Name:

Hexamethylene diisocyanate, oligomers, reaction products with 3-methoxypropylamine

Cas Number:

162492-07-1

Molecular formula:

C38H69N9O9

IUPAC Name:

Hexamethylene diisocyanate, oligomers, reaction products with 3-methoxypropylamine

Specific details on test material used for the study:

CAS No. 162492-07-1; EINECS No:. 500-740-9; Batch No.: 1591ZG-101; Purity >98 %; Appearance: homogeneous white powder

In vitro test system

Test system:

human skin model

Remarks:

EpiDermTM tissues

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

The test system is a commercially available EpiDermTM-Kit, procured by MatTek. The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, 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 representing main lipid classes analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell culture inserts.

Origin:
EpiDermTM tissues were procured from MatTek In Vitro Life Science Laboratories, Bratislava.
Designation of the kit: EPI-212-SIT

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

One plate was used as negative control; each tissue was treated with 30 μL DPBS buffer, a nylon mesh was added in order to ensure sufficient contact with the tissue surface. One plate was used as positive control; each tissue was treated with 30 μL 5% SDS solution, a nylon mesh was added in order to ensure sufficient contact with the tissue surface.

One plate was used for treatment with the test substance:
For each plate, 3 tissues were used. The tissues were wetted with 25 μL DPBS buffer before applying the test substance and spreading it to match the tissue size.
The following amounts were applied to the tissues:
Replicate 1: 25.3 mg
Replicate 2: 25.1 mg
Replicate 3: 25.6 mg

Duration of treatment / exposure:

1 h

Duration of post-treatment incubation (if applicable):

23 h and 30 min

Number of replicates:

3 replicates

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

99.4

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Results:
% Tissue viability (tissue 1): 99.9%
% Tissue viability (tissue 2): 99.9%
% Tissue viability (tissue 3): 98.4%
% Tissue viability (mean) 99.4%

Tissue Viability

The photometric absorbance of the negative controls is considered as 100%. For each replicate of test substance and positive control, tissue viability is calculated as % photometric absorbance compared with the mean of the negative controls:
% Tissue Viability = (OD replicate test substance resp. positive control x 100%) / OD negative controls

Acceptance of the results:

- The test substance is considered as non-irritant to skin.
- After the treatment, the relative absorbance values were reduced to 99.4%. This value is above the threshold for skin irritation (50%).
- The optical density of the negative control was well within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8.
- The positive control has met the acceptance criterion too, demonstrating the validity of the test system.
- Variation within replicates was within the accepted range for negative control, positive control and test substance (required: ≤ 18%).

Validity:
OD of negative control: 2.0 (Required: ≥ 0.8 and ≤ 2.8)
% Tissue viability of positive control SDS: 1.8% (Required: ≤ 20% of negative control)

SD of mean viability of the tissue replicates (%):
0.6% (negative control)
0.2% (positive control)
0.8% (test substance)
(Required: ≤ 18%)

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, after the treatment with the test substance, the relative absorbance values were reduced to 99.4 %. This value is above the threshold for skin irritation potential (50%). Therefore, the test substance is considered as non-irritant to skin in the Reconstructed Human Epidermis (RhE) test method.

Executive summary:

A study was conducted to determine the skin corrosion potential of the test substance using the Reconstructed Human Epidermis (RHE) test method according to OECD Guideline 439, in compliance with GLP. Three tissues of the human skin model EpiDermTM were treated with the test substance for 60 min. The substance was applied directly to each tissue and spread to match the tissue size (0.63 cm2). DPBS-buffer was used as negative control, 5% SDS solution as positive control. After treatment with the test substance or the positive / negative control, cell viability of the tissues was evaluated by addition of MTT. The absorbance values for the negative control were within the required acceptability range (OD = 2.0). The positive control showed clear irritating effects (elative absorbance was reduced to 1.8%). Variation within the tissue replicates was acceptable. Under the study conditions, after the treatment with the test substance, the relative absorbance values were reduced to 99.4 %. This value is above the threshold for skin irritation potential (50%). Therefore, the test substance is considered as non-irritant to skin in the Reconstructed Human Epidermis (RhE) test method (Andres, 2017).