1-hydroxyoctan-2-one

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16-25 Jan 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

This is a Klimisch 1 OECD 492 guideline study conducted on the registered substance 1-hydroxyoctan-2-one in accordance GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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:

1-hydroxyoctan-2-one

Cas Number:

7019-19-4

Molecular formula:

C8H16O2

IUPAC Name:

1-hydroxyoctan-2-one

Test material form:

liquid

Details on test material:

Batch 1203/16/100

Specific details on test material used for the study:

Batch 1081-67E

Test animals / tissue source

Species:

human

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

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

Duration of treatment / exposure:

30 minutes ± 2 minutes

Duration of post- treatment incubation (in vitro):

12 ± 2 minute 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)

Details on study design:

Prior to the study, the required compatibility checks confirmed that the test item interfered with MTT and therefore freeze killed controls (to assess direct MTT reduction) were included in the main test. There was no interference with the assay solvent.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

other: %viability

Remarks:

Corrected for freeze killed tissue control

Run / experiment:

1

Value:

12.571

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Results 2

Irritation parameter:

other: % tissue viability

Remarks:

Uncorrected for freeze killed tissue control

Run / experiment:

1

Value:

17.498

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Other effects / acceptance of results:

The viability of the MTT Interference Control (Freeze Killed Tissues) was 4.927%. Therefore, this value was subtracted from the uncorrected result of 17.498% tissue viability to yield the corrected result of 12.571% tissue viability

Any other information on results incl. tables

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

Condition	Tissue #	Raw data		Blank corrected data		Mean OD	% of viability
		aliquot 1	aliquot 2	aliquot 1	aliquot 2
NC	Tissue 1	2.145	2.159	1.977	1.991	1.984	102.347
	Tissue 2	2.085	2.092	1.917	1.924	1.920	99.071
	Tissue 3	2.093	2.065	1.925	1.897	1.911	98.581
PC	Tissue 1	1.25	1.226	1.082	1.058	1.070	55.193
	Tissue 2	0.889	0.905	0.721	0.737	0.729	37.601
	Tissue 3	0.784	0.779	0.616	0.611	0.613	31.642
TA1	Tissue 1	0.538	0.527	0.370	0.359	0.364	18.796
	Tissue 2	0.521	0.52	0.353	0.352	0.352	18.177
	Tissue 3	0.47	0.468	0.302	0.300	0.301	15.520

NC: negative control (sterile H₂O), PC: Positive control (neat Methyl Acetate), TA1: test item

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

Name	Code	mean of OD	SD of OD	Mean of viability (%)	SD of viability (%)	CV %	Classification
Sterile water	NC	1.938	0.040	100.000	2.05	2.048	No Category
Methyl Acetate	PC	0.804	0.237	41.479	12.245	29.521	No prediction
EXPINN PC17032	TA1	0.339	0.034	17.498	1.740	9.947	No prediction

Mean % viabilities obtained from additional tests to correct for MTT interference

Test Item	MTT Interference Control (FK): % viability	Initial % Viability	Corrected % Viability
TA1	4.927	17.498	12.571

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The percentage of viability obtained with test item 1 -hydroxyoctan-2 -one was 12.571%, therefore no prediction can be made

Executive summary:

In vitro Assessment of the eye irritation potential of 1 -hydroxyoctan-2 -one according to OECD guideline TG492 (Reconstructed human Cornea-like Epithelium [RhCE] test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage). The test item is applied directly to the cornea surface, providing a good model of “real life” exposure

 

This in vitro assessment assay predicts the acute eye irritation potential of a chemical by measurement of its cytotoxic effect on the EpiOcular™ corneal epithelial model. Cell viability is measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that is quantitatively measured after extraction from the skin tissues. Irritant test items are identified by their ability to decrease cell viability below defined threshold levels (below or equal to 60% for eye irritants ). If the viability is greater than 60%, the test item is classified as Non-Irritant (“no-label”, or UN GHS No Category).

 

The percentage of viability obtained with test item 1 -hydroxyoctan-2 -one was 12.571%, therefore no prediction can be made