Hydrocarbon waxes (petroleum), oxidized

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2nd February 2001 to 18th July 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

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:

- Storage: at room temperature and humidity.
- Physical appearance: Light brown semi-solid.
- TAN: 112-128

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Sgarlat's Rabbitry , Harvey's Lake, PA on 20/12/00 and 17/01/00.
- Eyes were examined before selection and assignment to the study.
- Age at study initiation: Animals were born on 08/10/00 and 02/11/00.
- Weight at study initiation: 2.1 - 2.4 kg.
- Housing: Individually in suspended cages.
- Diet (e.g. ad libitum): Purina Rabbit Chow Diet #5321, provided daily.
- Water (e.g. ad libitum): ad 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.
- Concentration (if solution): Used as supplied.

Observation period (in vivo):

Up to 21 days post exposure at the following time points; 1, 24, 48 and 72 hours, including days 7, 14 and 21.

Number of animals or in vitro replicates:

Three in total, 1 male and 2 females.

Details on study design:

TEST PROCEDURE:
- One female animal was tested initially, then based on this reaction a further two were tested.
- The test material was placed into the conjunctival sac of one eye, formed by gently pulling the lower eyelid from the eye. The lids were held together for one second before the animal was released.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not performed.

OBSERVATIONS:
-Bodyweight, was recorded before the exposure.
- Any additional signs and the general health of the animals was recorded throughout the study.

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

TOOL USED TO ASSESS SCORE:
- Sodium fluorescein dye was used at the following time points; 24, 48 and 72 hours and days 7, 14 and 21.
- Observations were made using a Mini-Maglit®.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 21 days in the female and 14 days in the male

Remarks on result:

other: Score of opacity

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 21 days in the female and 14 days in the male

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 21 days in the female and 14 days in the male

Remarks on result:

other: Score for redness

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

2.6

Max. score:

4

Reversibility:

fully reversible within: 21 days in the female and 14 days in the male

Irritant / corrosive response data:

Corneal opacity, noted in 3/3 eyes, cleared by day 21. Iritis, noted in 2/3 eyes, cleared by day 7.
Conjunctival irritation, noted in 3/3 days, cleared by day 21.

Other effects:

An instance of diarrhoea was the only abnormal physical sign noted during the study.

One animal was sacrificed on day 11 due to an injury not concurrent with treatment, the animal was replaced. None of the results from the sacrificed animal have been used in the study. This replacement is not thought to have affected the results of the study.

Any other information on results incl. tables

Table 2. Ocular Response

Animal No. and Sex	Tissue	Reading	1 hr	24 hrs	48 hrs	72 hrs	day 7	day 14	day 21
Female 1	Cornea	Opacity	0	0	2	0	0	0	0
		Area	0	0	1	0	0	0	0
	SubTotal		0	0	10	0	0	0	0
	Iris		0	0	0	0	0	0	0
	SubTotal		0	0	0	0	0	0	0
	Conjunctiva	Redness	2	2	2	2	1	1	0
		Chemosis	3	3	3	2	2	0	0
		Discharge	2	2	2	2	1	0	0
	SubTotal		14	14	14	12	8	2	0
	Total		14	14	24	12	8	2	0
Male 1	Cornea	Opacity	0	2	2	2	0	0
		Area	0	1	1	1	0	0
	SubTotal		0	10	10	10	0	0
	Iris		1	0	0	0	0	0
	SubTotal		5	0	0	0	0	0
	Conjunctiva	Redness	2	2	2	2	1	0
		Chemosis	3	3	2	2	0	0
		Discharge	2	2	2	1	0	0
	SubTotal		14	14	12	10	2	0
	Total		19	24	22	20	2	0
Female 2	Cornea	Opacity	0	0	2	2	2	2	0
		Area	0	0	2	2	2	1	0
	SubTotal		0	0	20	20	20	10	0
	Iris		0	1	1	1	0	0	0
	SubTotal		0	5	5	5	0	0	0
	Conjunctiva	Redness	2	2	2	2	2	0	0
		Chemosis	3	3	3	3	2	0	0
		Discharge	2	2	2	1	0	0	0
	SubTotal		14	12	14	12	8	0	0
	Total		14	19	39	37	28	10	0

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the test material was determined to be an eye irritant. Three out of three animals scored ≥ 2 for conjunctival redness and chemosis, which were observed to be fully reversible within 21 days. The material tested in this study has a TAN range of 112-128 and is classified as Eye Irritation Category 2 according to EU CLP Regulation No. 1272/2008.

Executive summary:

In a GLP compliant eye irritation study, the ocular reaction caused by exposure to the test material was determined according to EPA OPPTS 870.2400. Three rabbits were exposed to the unchanged test material and observed over a 21 day period. Three out of three animals scored ≥ 2 for conjunctival redness and chemosis, which were observed to be fully reversible within 21 days. The material tested in this study has a TAN range of 112-128 and based upon these results is classified as Eye Irritation Category 2 according to EU CLP Regulation No. 1272/2008. Corneal opacity and iritis were also observed in the test animals; however these symptoms were not severe enough to contribute to the classification.