Hydrocarbon waxes (petroleum), oxidized, Me esters, calcium salts

Endpoint summary

Description of key information

Skin irritation = Not irritating, male/female rabbit, EPA OPPTS 870.2500, Hoff 2001
Eye irritation = Not irritating, male rabbit, EPA OPPTS 870.2400, Cerven 2001

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 December 2000 to 15 December 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted 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. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sgarlat's Rabbitry, Harvey's Lake, PA
- Weight at study initiation: 2.2 - 2.6 kg
- Housing: individually in suspended wire cages. Bedding was placed underneath the cages and changed at least three times per week.
- Diet: Fresh Purina Rabbit Chow (Diet #5321) was provided daily
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Animals were kept in a temperature-controlled room with a 12 hour light/dark cycle.

IN-LIFE DATES: From 29 November 2000 to 15 December 2000

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

72 hours following patch removal

Number of animals:

3 (2 males, 1 female)

Details on study design:

TEST SITE
- Area of exposure: on the day prior to exposure, an area of approximately 10 x 15 cm on the dorsal trunk of each animals was clipped free of hair. 0.5 mL test material was applied to the test site.
- Type of wrap if used: the test material was covered with a 2.5 x 2.5 cm, 4 ply, gauze patch which was secured with non-irritating tape. The torso was wrapped with plastic in a semi-occlusive manner and secured with non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): at the end of the exposure period the wrappings were removed. Residual test material was removed from the test sites by gentle washing with distilled water prior to scoring for dermal reaction.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
The Draize scale, presented as Table 1 in the field "Any other information on material and methods incl. tables" was used to assess the degree of erythema and oedema at the application sites approximately 1 hour and 1, 2 and 3 days after the removal of the dressings. Any other clinical signs were also noted. Bodyweights were measured and recorded immediately before dose administration and at the 72 hour examination.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

0.56

Max. score:

2

Reversibility:

fully reversible within: 3 days

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

0.22

Max. score:

4

Reversibility:

fully reversible within: 2 days

Irritant / corrosive response data:

Erythema was barely perceptible at 60 minutes following patch removal, barely perceptible to well defined at 24 hours, absent to barely perceptible at 48 hours, and absent at 72 hours. Oedema was absent to barely perceptible at 60 minutes and 24 hours following patch removal, and absent at 48 and 72 hours.

Other effects:

There were no abnormal physical signs noted during the observation period. The dose site on one animal was stained yellow throughout the 72 hour observation period.
All bodyweight changes were normal.

Table 2: Results

Sex	M	M	F
Pre-test bodyweight (kg)	2.5	2.6	2.2
Terminal bodyweight (kg)	2.8	2.8	2.2
Time after patch removal (h)	Erythema
1	1a	1a	1a
24	1	1	2a
48	0	0	1a
72	0	0	0a
	Edema
1	1	1	0
24	1	0	1
48	0	0	0
72	0	0	0
	Systemic observations
1	A	A	A
24	A	A	A
48	A	A	A
72	A	A	A

A = Normal

a = dose site stained yellow

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant in accordance with EU criteria. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The skin irritation potential of the test material was determined in accordance with the standardised guideline EPA OPPTS 870.2500.

Three New Zealand White rabbits received a single four hour semi-occluded application of 0.5 mL of the test material and were assessed for the following 3 days for any signs of skin irritation. Erythema was barely perceptible at 60 minutes following patch removal, barely perceptible to well defined at 24 hours, absent to barely perceptible at 48 hours and absent at 72 hours. Oedema was absent to barely perceptible at 60 minutes and 24 hours following patch removal, and absent at 48 and 72 hours. There were no abnormal physical signs noted during the observation period and all bodyweight changes were normal.

Under the conditions of the test, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant in accordance with EU criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 December 2000 to 22 December 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted 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. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Millbrook Breeding Labs, Amherst, MA, USA
- Weight at study initiation: 2.4 - 2.7 kg
- Housing: individually in suspended cages. Bedding was placed underneath cages and changed three times per week.
- Diet: Fresh Purina Rabbit Chow (Diet #5321) was provided daily
- Water: ad libitum
- Acclimation period: at least 5 days
- Health: all animals were found to be free from evidence of ocular irritation or abnormalities before being assigned to the study

ENVIRONMENTAL CONDITIONS
- Animals were placed in a temperature controlled room which had a 12 hour light/dark cycle

IN-LIFE DATES: From: 29 November 2000 To: 22 December 2000

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

- Dose administration: Approximately 0.1 mL of the test material was placed by syringe into the conjunctival sac which was formed by gently pulling the lower eyelid away from the eye. After instillation, the lids were held together for approximately one second to insure adequate distribution of the test material. One eye of each rabbit was dosed. The contralateral eye served as a control.

Observation period (in vivo):

Animals were observed for 72 hours post administration.

Number of animals or in vitro replicates:

One animal initially, followed by a further two animals once the irritation potential was fully assessed in the first animal.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Not reported

SCORING SYSTEM
- The reactions observed were scored 1, 24, 48 and 72 hours post administration, in accordance with the criteria of Draize. The scale for scoring ocular reactions is presented as Table 1 in the field "Any other information on materials and methods incl. tables".
- Bodyweights were recorded pre-test
- The general health of the animals was monitored at each observation time

TOOL USED TO ASSESS SCORE
- Sodium fluorescein dye was used at the 24 hour observation interval. These examinations were performed using a Mini-Maglite flashlight equipped with a high intensity bulb.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: not applicable - no reactions seen

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: not applicable - no reactions seen

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

other: discharge

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.22

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

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

Other effects:

There were no abnormal physical signs noted during the observation period.

Table 2: Results

Animal	Tissue	Reading	1 hour	24 hours	48 hours	72 hours
1 M	Cornea	Opacity	0	0	0	0
		Area	0	0	0	0
		1. Total	0	0	0	0
	Iris		0	0	0	0
		2. Total	0	0	0	0
	Conjunctiva	Redness	2	2	2	0
		Chemosis	1	1	0	0
		Discharge	2	1	0	0
		3. Total	10	8	4	0
	Total = 1 + 2 + 3		10	8	4	0
	Systemic observations		A	A	A	A
	Sodium fluorescein			0
	Pre-test bodyweight		2.4 kg
2 M	Cornea	Opacity	0	0	0	0
		Area	0	0	0	0
		1. Total	0	0	0	0
	Iris		0	0	0	0
		2. Total	0	0	0	0
	Conjunctiva	Redness	1	1	1	0
		Chemosis	1	0	0	0
		Discharge	1	0	0	0
		3. Total	6	2	2	0
	Total = 1 + 2 + 3		6	2	2	0
	Systemic observations		A	A	A	A
	Sodium fluorescein			0
	Pre-test bodyweight		2.7 kg
3 M	Cornea	Opacity	0	0	0	0
		Area	0	0	0	0
		1. Total	0	0	0	0
	Iris		0	0	0	0
		2. Total	0	0	0	0
	Conjunctiva	Redness	2	2	1	0
		Chemosis	1	1	1	0
		Discharge	2	1	0	0
		3. Total	10	8	4	0
	Total = 1 + 2 + 3		10	8	4	0
	Systemic observations		A	A	A	A
	Sodium fluorescein			0
	Pre-test bodyweight		2.5 kg

 

A = Normal

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the test material only elicited slight reactions in the animals during the course of the study that meant the test material does not require classification as an eye irritant in accordance with EU criteria. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The eye irritation potential of the test material was determined in accordance with the standardised guideline EPA OPPTS 870.2400. During the study, 0.1 mL of test material was applied into one eye of three New Zealand White rabbits; the animals were assessed for 72 hours to determine the grade of ocular reaction.

There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation, which was noted in all three treated eyes, had fully cleared by 72 hours.

Under the conditions of the study, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant in accordance with EU criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation

Key Study
In the key study, the skin irritation potential of the test material was determined in accordance with the standardised guideline EPA OPPTS 870.2500.

Three New Zealand White rabbits received a single four hour semi-occluded application of 0.5 mL of the test material and were assessed for the following 3 days for any signs of skin irritation. Erythema was barely perceptible at 60 minutes following patch removal, barely perceptible to well defined at 24 hours, absent to barely perceptible at 48 hours and absent at 72 hours. Oedema was absent to barely perceptible at 60 minutes and 24 hours following patch removal, and absent at 48 and 72 hours. There were no abnormal physical signs noted during the observation period and all bodyweight changes were normal.

Under the conditions of the test, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant in accordance with EU criteria.

 

Supporting Information

In the supporting study (Anonymous, 1980b), the skin irritation potential of the test material was determined in accordance with the Code of Federal Regulations, Title 16, Chapter IIc, paragraph 1500.41.

During the study six New Zealand Albino rabbits received a single 24 hour occluded application of 0.5 mL of test material to abraded and unabraded skin; following application, they were assessed for any signs of skin irritation over a 72 hour period. All of the animals tested had slight erythema on the abraded sites and two out of the six animals had slight erythema on the unabraded sites after 24 hours. No erythema was present in any animal after 72 hours. No oedema was noted in any animal on either the abraded or unabraded sites at any time.

Under the conditions of the study, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant in accordance with EU criteria.

 

Eye Irritation

The eye irritation potential of the test material was determined in accordance with the standardised guideline EPA OPPTS 870.2400.

During the study, 0.1 mL of test material was applied into one eye of three New Zealand White rabbits; the animals were assessed for 72 hours to determine the grade of ocular reaction.

There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation, which was noted in all three treated eyes, had fully cleared by 72 hours.

Under the conditions of the study, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant in accordance with EU criteria.

 

The available data is considered to be complete and the conclusion for both skin irritation and corrosion and eye irritation, not irritating, was taken forward for risk assessment.

Justification for selection of skin irritation / corrosion endpoint:
The study reported by Hoff was selected as the key study since it was conducted to GLP and in line with the standardised guideline EPA OPPTS 870.2500. As such, the study was assigned a reliability score of 1 according to the criteria of Klimisch (1997) and considered suitable as an accurate reflection of the test material.

Justification for selection of eye irritation endpoint:
Only one study is available. The study was conducted to GLP and in line with the standardised guideline EPA OPPTS 870.2400. As such, the study was assigned a reliability score of 1 according to the criteria of Klimisch (1997) and considered suitable as an accurate reflection of the test material.

Justification for classification or non-classification

Skin

In accordance with the criteria for classification as defined in Annex I, Regulation 1272/2008, the test material does not require classification for skin irritation.

Eye

In accordance with the criteria for classification as defined in Annex I, Regulation 1272/2008, the test material does not require classification for eye irritation.