N,N-diethyl-2-propynylamine

Endpoint summary

Description of key information

The test substance is corrosive to the skin (according to OECD 431).

The test substance is irritating to the eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 29 July 2016

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

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

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

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

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.

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

According to internal standard; White Vienna rabbits were used. Usually 50µl or mg of the test substance were applied to the conjunctival sac of one eye of 2 animals. The non- or saline-treated adjacent eye served as a control.

GLP compliance:

no

Specific details on test material used for the study:

Name of the test substance used in the study report: Golpanol DEP = 1-Diethylaminopropin

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

- body weight at start of treatment: 2.99-3.47 kg

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

ca. 0.05 mL

Duration of treatment / exposure:

single treatment

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

The test substance was not washed out.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24 h

Score:

1

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion

In a key study (BASF, 2018), skin corrosion potential of the test item was tested according to OECD 431. The potential of the test substance to cause dermal corrosion was assessed by a single topical application of 50 μL of the undiluted test substance to a reconstructed three-dimensional human epidermis model (EpiDerm™). Two EpiDerm™ tissues were incubated with the test substance for 3 minutes and 1 hour, each. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test-substance treated epidermal tissues is compared to that of negative control tissues. The quotient of both values indicates the relative tissue viability. The following results were obtained in the EpiDermTM skin corrosion test: The test substance is able to reduce MTT directly. Therefore, an additional MTT reduction control KC (freeze-killed control tissues) was introduced. The final mean viability of the tissues treated with the test substance determined after an exposure period of 3 minutes was 96.3%, and it was 5.1% after an exposure period of 1 hour. Based on the observed results and by applying the evaluation criteria ,it was concluded, that the test item shows a corrosion potential in the EpiDerm™ in vitro skin corrosion test under the test conditions chosen. Additionally, in a supporting study (BASF, 1976) where the test item was tested on Vienna White rabbits under occlusive conditions on a shaved area, slight redness with spots; slight necrosis (anemic) were observed after 20-hour application. After 8 days: slight desquamation and severe necrosis (leather-like) was observed. Further, in a supporting study (Carpenter, et al., 1974) where the test item was tested on rabbits, the following results were observed: Irritation on uncovered rabbit belly: 1 / 10. Supporting studies cannot be used for classification.

 

Eye irritation

In a key study (BASF, 1976) the test item was tested on Vienna White rabbits for 24 hours and observed for 8 days. The test substance was not washed out after the treatment and the test was performed in duplicates. The mean cornea opacity score was 3 (max score = 4), the mean conjuctive score was 1 (max score = 3) and the mean chemosis score was 1 (max score = 4). None of the irritation parameters were reversible. Further in (Carpenter et al., 1974), the test item was tested on rabbits and corneal injury resulted in 8 / 10 in rabbits.

 

Justification for classification or non-classification

Based on the available information the test substance is classified as Skin corr. 1 H314: Causes severe skin burns and eye damage and Eye Damage 1 H318: Causes serious eye damage, in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.