Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

April 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

This is an OECD 492 guideline study conducted in accordance with GLP using substance Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)

EC Number:

245-327-1

EC Name:

Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)

Cas Number:

22919-56-8

Molecular formula:

C8H16O2.C6H15NO3

IUPAC Name:

2-[bis(2-hydroxyethyl)amino]ethan-1-ol; octanoic acid

Test material form:

liquid: viscous

Test animals / tissue source

Species:

human

Test system

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

After pre-wetting tissues with 20µl PBS (sterile Dulbecco’s Phosphate Buffered Saline) for 30 minutes ± 2 minutes, a single, topical application of 50µl of neat test item or of reference items was applied to the surface of the EpiOcular(TM) models for 30 minutes ± 2 minutes.

Duration of treatment / exposure:

30 minutes ± 2 minutes

Duration of post- treatment incubation (in vitro):

12 minutes ± 2 minutes post-treatment immersion, and 2 hours ± 15 minutes post-treatment incubation, prior to the MTT endpoint

Number of animals or in vitro replicates:

Three tissues per condition (n=3).

Results and discussion

In vitro

Results

Irritation parameter:

other: % Tissue Viability

Run / experiment:

1

Value:

ca. 5.537

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: No predication can be made as the viability was below 60%.

Any other information on results incl. tables

Results Summary

Test Item	Percentage of viability	Classification (No category/No prediction)
Octanoic Acid Triethanolamine Salt	5.537%	No Prediction

Viability measurements after 30 minutes (± 2 minutes) of application and 2 hours (± 15 minutes) post-incubation of test and reference items.

Condition	Tissue #	Raw Data		Blank Corrected Data		Mean OD	% Viability
		Aliquot 1	Aliquot 2	Aliquot 1	Aliquot 2
NC	Tissue 1	1.933	1.937	1.778	1.782	1.780	98.234
	Tissue 2	1.843	1.864	1.688	1.709	1.699	93.736
	Tissue 3	2.106	2.119	1.951	1.964	1.958	108.030
PC	Tissue 1	0.973	0.94	0.818	0.785	0.802	44.233
	Tissue 2	0.848	0.852	0.693	0.697	0.695	38.355
	Tissue 3	0.806	0.798	0.651	0.643	0.647	35.706
TA1	Tissue 1	0.293	0.279	0.138	0.124	0.131	7.230
	Tissue 2	0.261	0.259	0.106	0.104	0.105	5.795
	Tissue 3	0.222	0.218	0.067	0.063	0.065	3.587

Mean and SD of viability measurements and of viability percentages after 30 minutes (± 2 minutes) of application and 2 hours (± 15 minutes) post-incubation.

Name	Code	Mean of OD	SD of OD	Mean of viability (%)	SD of viability (%)	CV %	Classification
Sterile water	NC	1.812	0.132	100.000	7.31	7.309	No-Category
Methyl Acetate	PC	0.715	0.079	39.432	4.364	11.067	No Prediction
Octanoic Acid Triethanolamine Salt	TA1	0.100	0.033	5.537	1.835	33.136	No Prediction

  Evaluation of the Results - Results were checked against the following acceptance criteria.

	Description	Actual values	PASS/FAIL
Acceptance criterion 1	The mean OD570of the negative control(treated with sterile water) tissues is > 0.8 and < 2.5.	1.812	PASS
Acceptance criterion 2	The mean of the positive control relative percentage viabilityis below 50% of negative control viability after 30 minutes± 2 minutes exposure.	39.432%	PASS
Acceptance criterion 3	The SD of % viability between three tissue replicates should not exceed 18%in the same run (for negative and positive control tissues and tissues of test items).	NC: 7.31% PC: 4.364% TA1: 1.835%	PASS

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The percentage of viability obtained with test item Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) was 5.537%, therefore no prediction can be made.

Executive summary:

The eye irritation potential of Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) was assessed in vitro according to OECD Test Guideline 492 (Reconstructed human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage).

 

Prior to this study the required compatibility checks confirmed that the test item, Octanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1) did not interfere with MTT, and following the study all results were checked and met the acceptance criteria.

 

·        A test item is considered as “No Category”, i.e. not requiring a warning label in the European chemical classification systems for eye irritation or serious eye damage (UN GHS No Category), if the eye model viability after exposure and post-treatment incubation is >60%.

 

·        If the mean percent tissue viability after exposure and post-exposure incubation is ≤60%, no prediction can be made.

 

After 30 ± 2 minutes exposure on the surface of EpiOcular^™reconstructed ocular epithelium and 2 hours ± 15mins post-incubation time, the viability of the tissues was assessed and compared to a negative control.The percentage of viability obtained with test itemOctanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)was 5.537%, therefore no prediction can be made.