3,5-bis(2,4-dimethylcyclohex-3-en-1-yl)polyheterocycle

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to: INVITTOX Protocol No. 47: HET-CAM Test.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: INVITTOX Protocol No. 47: HET-CAM Test. http://ecvam-sis.jrc.it

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test animals / tissue source

Species:

other: HET-CAM

Strain:

not specified

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Approx. 100 mg of the neat test item were applied onto the CAM

Duration of treatment / exposure:

5 min

Details on study design:

Approx. 100 mg of the neat test item were applied onto the CAM in order to cover at least 50% of the CAM. The CAM was exposed to the test item for 5 minutes. During this 5 minutes reactions on the blood vessels in and under the CAM in close proximity to the test item were monitored. The CAMs of three eggs were treated with the test item. In addition to the test item a negative control (0.9% NaCl w/v)), two positive controls (1% SDS solution in deionised water and 0.1 N NaOH), and a reference item (5% (w/v) Texapon) were tested.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean (5 min. exposure)

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Results with Sa 57, as well as negative and positive controls, and the reference item

Test Group	Time until Haemorrhage [s]	Time until Lysis [s]	Time until Coagulation [s]	Irritancy Index	Mean Irritancy Index
Negative Control	301	301	301	0.00	0.00
	301	301	301	0.00
	301	301	301	0.00
Positive Control 0.1 N NaOH	10	45	17	19.34	19.26
	9	41	16	19.48
	11	57	20	18.96
Positive Control 1% SDS	13	30	301	11.12	11.11
	14	28	301	11.15
	14	32	301	11.06
Reference Item 5% AS Texpon	17	35	301	10.94	10.78
	22	47	301	10.58
	20	38	301	10.82
Test Item	301	301	301	0.00	0.00
	301	301	301	0.00
	301	301	301	0.00

Mean irritancy index of 3 eggs: Test Item = 0.00

Mean irritancy index of 3 eggs: Positive Control (Sodium dodecyl sulphate) = 11.11

                                                        Positive Control (Sodium hydroxide solution) = 19.26

                                                        Reference Item (Texapon) = 10.78

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

other: INVITTOX Protocol No. 47: HET-CAM Test. http://ecvam-sis.jrc.it

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item Sa 57 does not possess any irritating potential.

Executive summary:

This in vitro study was performed to assess the irritating potential of Sa 57 by detection of damages in blood vessels under the chorioallantoic membrane of 9 day incubated chicken eggs.

Each 300 µL of the test item, the positive and the negative controls, and the reference item were applied to three eggs each such that at least 50 % of the membrane was covered. During the observation period for 300 seconds any lesions in close proximity to the covered membrane were monitored and recorded. The observation time was 5 minutes at room temperature.

Physiological sodium chloride solution (0.9 % (w/v) in deionised water) was used as negative control.

The negative control showed no irritating effect on the blood vessels under the membrane. The irritancy mean index is 0.00.

A 1 % solution of SDS and 0.1 NaOH were used as positive controls.

The positive controls induced a severe irritation on the blood vessels. The calculated mean irritancy indices are 11.11 for 1 % SDS and 19.26 for 0.1 N NaOH. The reference itemTexapon ASV Spezial (5 % AS) was chosen because of its known eye irritating properties and induced a severe irritation on the blood vessels. The calculated mean irritancy index is 10.78.

The mean irritancy indices of the controls are well comparable with the historical control values and are within the acceptance criteria. The result of the reference item demonstrated a correct interpretation. Therefore, it was concluded that the test was valid.

No irritating effects were observed during 5 min incubation with Sa 57. The calculated irritancy mean index is 0.00.