Di-tert-butyl 1,1,4,4-tetramethyltetramethylene diperoxide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 September 1994 - 27 October 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well-conducted and documented study performed under GLP; however, no CoA for the test material is included in the report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4500 (Acute Eye Irritation)

Deviations:

yes

Remarks:

One mortality check was not conducted and one was conducted but not documented. However, no mortality occurred during the study and the occurences are considered to have had no apparent impact on the outcome of the study.

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Di-tert-butyl 1,1,4,4-tetramethyltetramethylene diperoxide

EC Number:

201-128-1

EC Name:

Di-tert-butyl 1,1,4,4-tetramethyltetramethylene diperoxide

Cas Number:

78-63-7

Molecular formula:

C16H34O4

IUPAC Name:

2,5-bis(tert-butylperoxy)-2,5-dimethylhexane

Details on test material:

Chemical name: no data
Purity: no data
Lot No.: 0568422701
Receipt Date: September 9, 1994
Physical Description: Light yellow liquid
Storage Conditions: Room temperature
Expiration Date: February 15, 1995

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Adult, New Zealand White rabbits were received at SLS from Myrtle's Rabbitry, Thompson Station, TN. Upon receipt, plastic ear tags displaing unique identification numbers were used to individually identify the animals. Cage cards displaying at least the study number, animal number and sex were affixed to each cage. The animals were housed individually in suspsended stainles steel cages. All housing and care were based on the standards recommended by the Guide for the Care and Use of Laboratory Animals.

The animal room temperature and relative humidity ranges were 68 - 70 °F and 46 - 59%, respectively. Environmental control equipment was monitored and adjusted as necessary to minimize fluctuations in the animal room environment. Light timers were set to maintain a 12-hour light/12-hour dark cycle. There were ten to twelve air changes in the animal room per hour. the animal room temperature and relative humidity were recorded a minimum of once daily.

Purina Certified Rabbit chow #5322 was provided ad libitum to the animals throughout the study. the lot number and expiration date of each batch of diet used during the study were recorded. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Dietary limitation for various environmental contaminants, including heavy metals. pesticides, polychlorinated biphenyls and total aflatoxin are set by the manufacturer. Within these limits, contaminants which may have been present were not expected to compromise the purpose of this study. Results of the dietary analyses (Certificates of Analysis) are provided by the manufacturer for each lot of diet. These are maintained by SLS.

Municipal tap water treated by reverse osmosis or deionization (back-up system) was available to the animals ad libitum throughout the study. The purified water was supplied by an automatic watering system. Monitoring of the drinking water for contaminants was conducted by SLS and the records are available for inspection. Within generally accepted limits, contaminants which may have been present were not expected to compromise the purpose of this study.

Upon receipt, animals were examined, identified with plastic ear tags and then quarantined for a minimum of five days. The animals chosen for study use were arbitrarily selected from healthy stock animals to avoid potential bias. All animals received a detailed pretest observation prior to dosing. Only healthy animals were chosen for study use. Females were nulliparous and nonpregnant.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other:

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

Once

Observation period (in vivo):

The eyes were macroscopically examined with the aid of an auxiliary light source for signs of irritation at 1, 24, 48 and 72 hours after dosing according to the Ocular Grading System presented in Appendix B which is based on Draize. Following macroscopic observations at the 24 hour scoring interval, the fluorescein examination procedure was repeated on all test and control eyes and any residual test article was gently rinsed from the eye at this time (if possible) using 0.9% physiological saline. If positive fluorescein dye retention was noted at 24 hours, a fluorescein exam was conducted on the affected eyes at each subsequent interval until a negative response was obtained.

Number of animals or in vitro replicates:

six

Details on study design:

The test article was instilled into the conjunctival sac of the right eye of each animals after gently pulling the lower lid away from the eye. Following instillation, the eyelids were gently heald together for approximately one second in order to limit test article loss and the animal was returned to its cage. The contralateral eye remained untreated to serve as a control.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Remarks:

(redness + swelling + discharge) x 2

Basis:

mean

Remarks:

mean of all six animals

Time point:

other: 1 hour

Score:

5

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 2

Irritation parameter:

conjunctivae score

Remarks:

(redness + swelling + discharge) x 2

Basis:

mean

Remarks:

mean of all six animals

Time point:

other: 24 hours

Score:

2.33

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness + swelling + discharge) x 2

Basis:

mean

Remarks:

mean of all six animals

Time point:

other: 48 hours

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 4

Irritation parameter:

conjunctivae score

Remarks:

(redness + swelling + discharge) x 2

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 5

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: all

Score:

0

Max. score:

0

Results 6

Irritation parameter:

iris score

Basis:

mean

Time point:

other: all

Score:

0

Max. score:

0

Irritant / corrosive response data:

Exposure to the test article produced conjunctivitis (redness, swelling and/or discharge) in 6/6 test eyes at the 1 hour scoring interval. The conjunctival irritation resolved completely in all animals by the 72 hour scoring interval.
Normal background positive fluorescein dye retention was observed in 1/6 control eyes (stippling) at the 24 hour scoring interval. This finding was resolved by 48 hours and was not considered significant. No corneal opacityI iritis or conjunctivitis was observed in the control eyes.

Any other information on results incl. tables

Summary of ocular lesions

Anim. No.	Effect	Hours	Days after application					Mean score cornea Days 1/2/3	Mean score iritis Days 1/2/3	Mean score redness Days 1/2/3	Mean score chemosis Days 1/2/3
		1	1	2	3	7	14
51369/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 1	0 0 1 1 0	0 0 0 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0.33	0
51370/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 1	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0	0
51364/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 1	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0.33	0
51337/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 0	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0.33	0
51362/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 0	0 0 1 0 0	0 0 1 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0.66	0
51275/F	Cornea Iris Redness Chemosis Discharge	0 0 1 1 0	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	- - - - -	- - - - -	0	0	0.33	0
Mean all anim.								0	0	0.33	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Slight conjunctival effects were noted in this study; however, they were not severe enough to warrant classification.
Based on these results, the test substance does not have to be classified and has no obligatory labelling requirement for eye irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2017) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Executive summary:

The potential eye irritant and/or corrosive effects of the substance were evaluated on New Zealand White rabbits. Each of six rabbits received a 0.1 ml dose of the test article in the conjunctival sac of the right eye. The conralateral eye of each animal remained untreated and served as a control. Test and control eyes were examined for signs of irritation for up to 72 hours following dosing.

Exposure to the test article produced conjunctivitis (redness, swelling and/or discharge) in 6/6 test eyes at the 1 hour scoring interval. The conjunctival irritation resolved completely in all animals by the 72 hour scoring interval.