Alcohols, C18-unsatd and ethoxylated C18-unsatd., phosphates (5 moles ethoxylation)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 06, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

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)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, C18-unsatd and ethoxylated C18-unsatd., phosphates (5 moles ethoxylation)

Molecular formula:

C18H37O4P (representative of phosphate monoesters; i.e., mono- C18-unsatd. PSE) C28H57O9P1 (representative of ethoxylated phosphate monoesters; i.e., mono- C18-unsatd. AE5 PSE) C36H71O4P1 (representative of phosphate diesters; i.e., di- C18-unsatd. PSE) C46H91O8P1 (representative ethoxylated phosphate diesters; di- C18-unsatd. PSE and C18-unsatd. AE5 PSE) C82H161O21P1 (representative ethoxylated phosphate diesters; di- C18-unsatd. AE5 PSE)

IUPAC Name:

Alcohols, C18-unsatd and ethoxylated C18-unsatd., phosphates (5 moles ethoxylation)

Test material form:

liquid

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Source of Bovine Eyes
Eyes from adult cattle (typically 12 to 60 months old) 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 µg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.75 mL

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

32 ± 1 ºC for 120 minutes

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

ca. 52.3

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: No prediction can be made

Other effects / acceptance of results:

Criteria for an Acceptable Test
The positive control In Vitro Irritancy Score was within the range of 31.6 to 58.7. The positive control acceptance criterion was therefore satisfied. The negative control gave opacity of ≤3.0 and permeability ≤0.077. The negative control acceptance criteria were therefore satisfied.

Any other information on results incl. tables

Results

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in below table:

Treatment	Cornea Number	Opacity					Permeability (Optical Density)		In Vitro Irritancy Score
		Pre-Treatment	Post-Treatment	Post Incubation	Post-Incubation - Pre‑Treatment	Corrected Value		Corrected Value
Negative Control #	1	3	2	2	0		0.002
	2	2	3	2	0		0.002
	3	3	2	2	0		0.003
	Mean				0.0*		0.002		0.0
Positive Control #	4	0	27	30	30	30.0	1.805	1.803
	5	3	35	32	29	29.0	1.133	1.131
	6	0	27	29	29	29.0	1.043	1.041
	Mean					29.3		1.325	49.2
Test Substance	10	0	23	45	45	45.0	0.306	0.304
	11	1	37	55	54	54.0	0.088	0.086
	12	0	27	51	51	51.0	0.063	0.061
	Mean					50.0		0.150	52.3

* = Mean of the post-incubation-pre‑treatment values                           

# = Control data shared with Envigo - Shardlow study number BS88JJ

Corneal Epithelium Condition

The condition of each cornea is given in below table:

 

Treatment	Cornea Number	Observation
		Post Treatment	Post Incubation
Negative Control #	1	Clear	Clear
	2	Clear	Clear
	3	Clear	Clear
Positive Control #	4	Cloudy	Cloudy
	5	Cloudy	Cloudy
	6	Cloudy	Cloudy
Test Substance	10	Cloudy – with some test substance remaining	Cloudy – with some test substance remaining
	11	Cloudy – with some test substance remaining	Cloudy – with some test substance remaining
	12	Cloudy – with some test substance remaining	Cloudy – with some test substance remaining

 

The corneas treated with the test substance were cloudy post treatment and post incubation with some test substance remaining adhered to the corneas. The corneas treated with the negative control substance were clear post treatment and post incubation. The corneas treated with the positive control substance were cloudy post treatment and post incubation.

 

In Vitro Irritancy Score

The In Vitro irritancy scores are summarized as follows:

Treatment	In Vitro Irritancy Score
Test Substance	52.3
Negative Control	0.0
Positive Control	49.2

 

Criteria for an Acceptable Test

The positive control In VitroIrritancy Score was within the range of 31.6 to 58.7. The positive control acceptance criterion was therefore satisfied. The negative control gave opacity of ≤3.0 and permeability ≤0.077. The negative control acceptance criteria were therefore satisfied.

 

Conclusion

Based on the study results, the study author concluded that no prediction of eye irritation could be made.

Applicant's summary and conclusion

Interpretation of results:

other: No prediction can be made based on EU CLP criteria

Conclusions:

Under the study conditions, no prediction could be made about eye irritation potential of the test substance.

Executive summary:

An in vitro study was conducted to determine the eye irritation potential of the the test substance, 'mono- and di- C18-unsatd. PSE and C18-unsatd. AE5 PSE', using Bovine Corneal Opacity and Permeability (BCOP) method, according to OECD Guideline 437 and EU Method B.47, in compliance with GLP. The undiluted test substance and reference substances were applied to the test system for 10 minutes followed by an incubation period of 120 minutes. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). The test substance IVIS was 52.3, which is well below the corrosive limit of 55. Therefore no prediction can be made about eye irrtation potential of the test substance, based on EU CLP criteria. The positive control IVIS was within the range of 31.6 to 58.7 and the negative control gave opacity of ≤3.0 and permeability ≤0.077, therefore, the test was considered to have met all validity criteria. Under the study conditions, no prediction could be made about eye irritation potential of the test substance (Envigo, 2017).