N,N-diethyl-2-propynylamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 Dec 2016 to 29 Nov 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

adopted 29 July 2016

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.40 BIS: Methods for the determination of toxicity and other health effects: In Vitro Skin Corrosion: Human Skin Test Method; Official Journal of the European Union, No. L 142

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF SE Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany

Test material

Test material information

Constituent 1

Reference substance name:

N,N-diethyl-2-propynylamine

EC Number:

223-804-5

EC Name:

N,N-diethyl-2-propynylamine

Cas Number:

4079-68-9

Molecular formula:

C7H13N

IUPAC Name:

diethyl(prop-2-yn-1-yl)amine

Specific details on test material used for the study:

- Name of test substance: Golpanol DEP
- Test-substance No.: 16/0439-1
- Batch identification: 201512251
- Purity: 98.3 corrected area-%
- Physical state / color: Liquid / colorless to yellowish, clear
- Homogeneity: The test substance was homogeneous by visual inspection.
- Storage stability: The stability under storage conditions over the study period was guaranteed by the sponsor
- Storage conditions: Room temperature

In vitro test system

Test system:

human skin model

Remarks:

EpiDermTM Reconstructed Human Epidermis

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: 24 EPI-200 tissues (reconstructed epidermis): surface 0.6 cm2 cultured in Millicells® 1 cm

Source strain:

not specified

Vehicle:

unchanged (no vehicle)

Details on test system:

MATERIALS AND TECHNICAL EQUIPMENT
- Laminar flow bench: HERAsafe KS 18 (Thermo ELECTRON CORPORATION)
- CO2 incubator: Heraeus BBD 6220
- Incubation conditions: 37°C ± 1°C, 5% ± 1% CO2, 90% ± 5% humidity
- Spectrophotometer: SunriseTM Absorbance Reader. For the determination of the optical density of colored extracts. Measurement using a filter wavelength 570 nm without reference filter
- EpiDerm™ 200 kit: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia containing: 24 EPI-200 tissues (reconstructed epidermis): surface 0.6 cm² cultured in Millicells® of 1 cm diameter
- Tissue for MTTreduction control: EPI-200 tissue that is killed by freezing at –20°C
- Assay medium: EPI-100-ASY assay medium; MTT diluent: Dulbecco's modified eagle's medium (DMEM) based medium used for diluting MTT (MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia / Sigma, Germany)
- Wash buffer: Dulbecco's phosphate buffered saline (PBS), w/o Ca 2+, Mg 2+ (MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia and Biochrom, Germany)
- Detection agent: 3-[4.5-dimethylthiazol-2-yl]-2.5-diphenyltetrazolium bromide (MTT) (MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia / Sigma, Germany), 1.0 mg / mL MTT diluent
- Extracting agent: Isopropanol p.a.

THREE DIMENTIONAL HUMAN EPIDERMIS MODEL:
- The EpiDermTM model consists of normal, 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 arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm diameter) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.
- Tissue model: EPI-200
- Tissue Lot Number: 23385
- Origin: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

- Fifty microliter (50 μL) of the undiluted liquid test substance was applied using a pipette.
- Control tissues were concurrently treated with 50 μL of negative control or with 50 μL of positive control or test substance.

Duration of treatment / exposure:

3 minutes at room temperature or 1 hour in the incubator

Number of replicates:

Two tissues per exposure time

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

Exposure period 3 min

Run / experiment:

mean values

Value:

96.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Remarks:

Exposure period 1 h

Run / experiment:

mean

Value:

5.1

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

Due to the ability of the test substance to reduce MTT directly, KC tissues were applied in parallel. The results of the KC tissues indicate an increased MTT reduction (mean viability 0.35% of NC). Thus, for the test substance the final mean viability is given after KC correction.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (corrosive) based on GHS criteria