2-propen-1-amine, N-2-propen-1-yl-, hydrochloride

Endpoint summary

Description of key information

The in vitro skin irritation of 2-propen-1-amine, N-2-propen-1-yl, hydrochloride was determined in accordance with the OECD Guideline 439 during a GLP compliant study.  This in vitro risk assessment assay predicits the skin irritation potential of a chemical by measurement of its cytotoxic effect on the EpiDerm tissue model.

Prior to testing, the test substance was checked for interference with water and/or MTT.  No interference was identified.

Skin irritation of the test substance and controls was evaluated in triplicate.  After 15 minutes exposure on the surface of the EpiDerm resconstructed human epidermis and a 42 hour post-exposure incubation time, viability of the tissues was assessed and compared to the negative control.

The percentage viability obtained with 2-propen-1-amine, N-2-propen-1-yl, hydrochloride was 108.6%, therefore it was considered as non-irritant to the skin according to criteria laid down in OECD guideline 439.

The test item did not meet the criteria for classification as irritant according to Regulation (EC) No. 1272/2008.

 

An in vitro Bovine Corneal Opacity and Permeability (BCOP) assay was performed in line with OECD testing guideline 437.  The study was GLP compliant.

The undiluted test material was applied neat to three cattle corneas obtained from a abattori at a volume of 0.75 mL, after which each cornea was incubated at 32 oC for 10 minutes.  Opacity and permeability of corneas were measuring following exposure.

Negative and positive controls were considered valid.

No prediction could be made from the BCOP eye irritation study for 2-propen-1-amine, N-2-propen-1-yl, hydrochloride. Based on the information available from the skin irritation studies, a worst-case scenario prediction of GHS category 2 for eye irritation has been implemented.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 2021 to April 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

66% purity
Aqueous solution

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIN (RHE) TISSUE
Supplier: Episkin Laboratories, Lyon, France
Tissue model: 0.38 cm2 reconstrcuted spidermin of normal keratinocytes
Batch number: 21-EKIN-003
Date received: 19th February 2021

TEMPERATURE USED DURING TREATMENT
Temperature: 37oC

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
Viability: Optical Density (O.D) Values: 0.843 +/- 0.008 (mean and SD of MTT value of 3 tissues exposed to negative control). Acceptance criteria (>0.6 and <1.5) - pass

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST ITEM
- used as supplied (neat)
- Volume applied, 10 uL

NEGATIVE CONTROL
- used as supplied (neat)
-Volume applied, 10 uL

POSITIVE CONTROL
- prepared as a 5% w/v aqueous solution.
-volume aplied, 10 uL

Duration of treatment / exposure:

EXPOSURE PERIOD - 15 minutes

Duration of post-treatment incubation (if applicable):

Incubated at 37 oC for 42 hours

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean value

Value:

108.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
-Visible damage on test system: none reported
- Direct-MTT reduction: No direct reduction
-Colour interference with MTT: No colour interference

Interpretation of results:

GHS criteria not met

Conclusions:

2-Propen-1-amine, N-2-propen-1-yl-, hydrochloride did not reduce the tissue viability. The test substance does not meet the criteria for classification as irritant to the skin according to Regulation (EC) No. 1272/2008.

Executive summary:

The in vitro skin irritation of 2-propen-1-amine, N-2-propen-1-yl, hydrochloride was determined in accordance with the OECD Guideline 439 during a GLP compliant study.  This in vitro risk assessment assay predicits the skin irritation potential of a chemical by measurement of its cytotoxic effect on the EpiDerm tissue model.

Prior to testing, the test substance was checked for interference with water and/or MTT.  No interference was identified.

Skin irritation of the test substance and controls was evaluated in triplicate.  After 15 minutes exposure on the surface of the EpiDerm resconstructed human epidermis and a 42 hour post-exposure incubation time, viability of the tissues was assessed and compared to the negative control.

The percentage viability obtained with 2-propen-1-amine, N-2-propen-1-yl, hydrochloride was 108.6%, therefore it was considered as non-irritant to the skin according to criteria laid down in OECD guideline 439.

The test item did not meet the criteria for classification as irritant according to Regulation (EC) No. 1272/2008.

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

Study period:

February 2021 to May 2021

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)

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Eyes from adult cattle (typically 12 to 60 months) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks' Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 ug/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST ITEM
-used neat
-volume applied, 0.75 mL

POSITIVE CONTROL
-used neat
-volume applied, 0.75 mL

NEGATIVE CONTROL
-used neat
-volume applied, 0.75 mL

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

120 minutes

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
The medium from both chambers of each holder was replaced with fresh complete EMEM. A pre-treatment opacity reading was taken for each cornea using a calibrated opacitometer. Three corneas were randomly allocated to the negative control. Three corneas were also allocated to the test item and three corneas to the positive control.

NUMBER OF REPLICATES
Triplcate

NEGATIVE CONTROL USED
Sodium chloride 0.9%w/v

POSITIVE CONTROL USED
Ethanol

APPLICATION DOSE AND EXPOSURE TIME
A volume of 0.75 mL of the test substance was applied to each of the three corneas followed by a 10 minute incubation at 32 oC.

POST-INCUBATION PERIOD: yes

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3
- POST-EXPOSURE INCUBATION: 120 minutes

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Labtech LT-4500 microplate reader
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean score

Value:

28.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: The corneas treated with the test item were cloudy post-treatment and post-incubation. The corneas treated with the negative control were clear post-treatment and post-incubation. The corneas treated with the positive control item were cloudy post-treatment and post-incubation.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The test was valid as the negative control produced an In Vitro Irritancy Score of 0.1 which is less than or equal to the upper limit for background opacity and permeability values calculated from the previous 12 months data for the testing facility.
- Acceptance criteria met for positive control: The test is normally acceptable if the positive control produced an In Vivo Irritancy Score which fell within two standard deviations of the historical mean collated during the previous 12 months for the testing facility. The positive control IVIS (30.3) was marginally below the normally accepted limit of 33.7-61.4. However, the data obtained was considered acceptable in demonstrating the sensitivity of the system. This IVIS score obtained does not qualify as an outlier (<26.8) and the data will be added to the historical control data, and is therefore accepted as a valid result.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Remarks:

According to the UN GHS Classification for the test item 2-propen-1-amine, N-2-propen-1-yl, hydrochloride, no prediction of eye irritation could be made under the conditions of the test. As the skin irritation study produced a non-irritant result, a worst-case scenario of GHS category 2 for eye irritation has been determined.

Conclusions:

No prediction could be made from the BCOP eye irritation study for 2-propen-1-amine, N-2-propen-1-yl, hydrochloride. Based on the information available from the skin irritation studies, a worst-case scenario prediction of GHS category 2 for eye irritation has been implemented.

Executive summary:

An in vitro Bovine Corneal Opacity and Permeability (BCOP) assay was performed in line with OECD testing guideline 437.  The study was GLP compliant.

The undiluted test material was applied neat to three cattle corneas obtained from a abattori at a volume of 0.75 mL, after which each cornea was incubated at 32 oC for 10 minutes.  Opacity and permeability of corneas were measuring following exposure.

Negative and positive controls were considered valid.

No prediction could be made from the BCOP eye irritation study for 2-propen-1-amine, N-2-propen-1-yl, hydrochloride. Based on the information available from the skin irritation studies, a worst-case scenario prediction of GHS category 2 for eye irritation has been implemented.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Justification for classification or non-classification

No adverse effects were observed during the skin irritation study and therefore the substance did not meet the requirements for classification.

 

Adverse effects were observed during the eye irritation study and as such was classified as a category 2 eye irritant.