Paraffin waxes and Hydrocarbon waxes, oxidized, lithium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3rd December 2001 to 6th December 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hydrocarbon waxes (petroleum), oxidized

EC Number:

265-205-1

EC Name:

Hydrocarbon waxes (petroleum), oxidized

Cas Number:

64743-00-6

Molecular formula:

too complex

IUPAC Name:

Hydrocarbon waxes (petroleum), oxidized

Test material form:

other: greasy solid

Details on test material:

- Physical appearance: Light brown semi-solid.
- Storage condition of test material: Room temperature and humidity.
- TAN: 24 - 40

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Millbrook Breeding Lads, Amherst, MA on 31/10/01.
- Only healthy animals free from ocular damage were selected for this study.
- Age at study initiation: Born on 02/09/01.
- Weight at study initiation: 3.2 - 3.4 kg.
- Housing: Individually in suspended cages.
- Diet (e.g. ad libitum): Purina Rabbit Chow, diet # 5321, supplied by Purina Mills, Inc. Feed daily.
- Water (e.g. ad libitum): as libitum.
- Quarantine period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Controlled.
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The contralateral eye served as a control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml/rabbit.

Observation period (in vivo):

- Total of 72 hours.

Number of animals or in vitro replicates:

- Three female animals.

Details on study design:

METHOD
- The test material was placed into the conjunctival sac, formed by gently pulling the lower eyelid away from the eye. The lid was held closed for a second, before the animal was released.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No.

SCORING SYSTEM: Draize scale (1977), which can be seen in table 1.

OBSERVATIONS
-Time points: 1, 24, 48 and 72 hours.
- General health was recorded at each time point.
- Body weights were recorded before exposure.

TOOL USED TO ASSESS SCORE:
- A Mini-Maglite® was during ocular examinations.
- Sodium fluorescein dye was used at 24 hours.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: No reaction observed

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Max. score:

2

Remarks on result:

other: No reaction observed

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: mean of 24, 48 and 72 hours

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: mean of 24, 48 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: No reaction observed

Irritant / corrosive response data:

There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation, noted in 3/3 eyes, cleared within 72 hours.

Other effects:

- No abnormal physical signs were noted during the observation period.

Any other information on results incl. tables

Table 2. Results for Ocular Damage.

Animal No.	Tissue	Reading	1 hr	24 hrs	48 hrs	72 hrs	Mean Score from 24, 48 and 72 hours
1	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0
	SubTotal		0	0	0	0
	Iris		0	0	0	0	0
	SubTotal		0	0	0	0
	Conjunctiva	Redness	2	0	0	0	0
		Chemosis	2	0	0	0	0
		Discharge	2	0	0	0
	SubTotal		12	0	0	0
	Total		12	0	0	0
2	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0
	SubTotal		0	0	0	0
	Iris		0	0	0	0	0
	SubTotal		0	0	0	0
	Conjunctiva	Redness	2	1	0	0	0.33
		Chemosis	2	0	0	0	0
		Discharge	2	0	0	0
	SubTotal		12	2	0	0
	Total		12	2	0	0
3	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0
	SubTotal		0	0	0	0
	Iris		0	0	0	0	0
	SubTotal		0	0	0	0
	Conjunctiva	Redness	2	1	1	0	0.66
		Chemosis	2	0	0	0	0
		Discharge	2	0	0	0
	SubTotal		12	2	2	0
	Total		12	2	2	0

Applicant's summary and conclusion

Interpretation of results:

not irritating

Conclusions:

Under the conditions of the test, the test material was determined to be non-irritating according to Regulation (EC) 1272/2008. The material tested in this study has a TAN range of 24-40. The observed ocular effects in the three animals were not severe enough to warrant classification. Initially animals displayed mild chemosis and discharge which completely cleared by 24 hours. Two of the three animals showed mild conjunctiva redness which was completely reversible within 72 hours.

Executive summary:

In a GLP compliant study performed according to the standardised guideline EPA OPPTS 870.2440, the ocular irritation caused by exposure to the test material was assessed in three rabbits. They were exposed to 0.1 ml of the test material using their contralateral eye as a control. Initially some mild irritation (chemosis and discharge) was observed, however this did not persist past 1 hour. Conjunctiva redness was the parameter which persisted past 1 hour and was assigned an average score of 0.33 according to the Draize scale (1977), this was completely reversible within 72 hours. Under the conditions of this study the test material (which has a TAN range of 24 - 40) does not require classification as an eye irritant in line with Regulation (EC) 1272/2008.