Reaction mass of 2,4-bis(xylylazo)resorcinol and 2,4-bis[(4-dodecylphenyl)azo]resorcinol and 2-[(4-dodecylphenyl)azo]-4-(2,4-xylylazo)resorcinol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5-28-1993 to 5-31-1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: EPA/OECD guideline study performed in accordance with GLP; exact details of test material (certificate of analysis, Characterisation) are not included in the report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not specified

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2,4-bis(xylylazo)resorcinol and 2,4-bis[(4-dodecylphenyl)azo]resorcinol and 2-[(4-dodecylphenyl)azo]-4-(2,4-xylylazo)resorcinol

EC Number:

915-586-1

Molecular formula:

Variable; substance is a UVCB.

IUPAC Name:

Reaction mass of 2,4-bis(xylylazo)resorcinol and 2,4-bis[(4-dodecylphenyl)azo]resorcinol and 2-[(4-dodecylphenyl)azo]-4-(2,4-xylylazo)resorcinol

Test material form:

other: Semi-solid

Details on test material:

The test material was a red colored semi-solid, identified as Automate Yellow 8 Petroleum. The test material was administered undiluted. Records of the test article stability, purity, source and other data required by Federal Regulations will be maintained by the Sponsor.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

One group of six male New Zealand White rabbits were used for this study. The rabbits were obtained from Oak Hill, Holland, Michigan, and weighed between 2.24 and 2.85 kg. at study initiation.
The rabbits were identified by ear tags and were individually housed in stainless steel cages in a temperature, humidity, and light controlled room. The animals were maintained according to the recommendations contained in DHHS Publication No. 86-23 (NIH): Revised 1985, "Guide for the Care and Use of Laboratory Animals." They were conditioned for 5 days prior to study initiation. Purina Rabbit Chow (ca. 350 gm) was placed in the feeders each morning during the study. Tap water from an automatic watering system was provided ad libitum from arrival until termination. Six animals were selected randomly from the acclimated colony and assigned to the test group.

Justification for Selection of Test System: The New Zealand Albino rabbit has been used extensively for eye irritation evaluations.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The contralateral eye served as the untreated control for each rabbit.

Amount / concentration applied:

0.1 g of undiluted test material.

Duration of treatment / exposure:

Single exposure.

Observation period (in vivo):

The treated eyes were examined and scored at 1, 24, 48 and 72 hours after treatment. A solution of 2% sodium fluorescein and ultraviolet light were employed to reveal possible corneal injury. Scoring was performed in accordance with the Draize Scale For Scoring Ocular Lesions (Table 1).

Number of animals or in vitro replicates:

6 animals.

Details on study design:

The test material (0.1 g) was flattened into a small wafer and was instilled into the conjunctival sac of the right eye of each rabbit, and the eyelids were held together for one second.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: Mean score for all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Results 2

Irritation parameter:

iris score

Basis:

animal: Mean score for all animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

not specified

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: Mean score for all animals

Time point:

24/48/72 h

Score:

0.28

Max. score:

3

Reversibility:

not specified

Results 4

Irritation parameter:

chemosis score

Basis:

animal: Mean score for all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Any other information on results incl. tables

Table 2: INDIVIDUAL AND GROUP MEAN BODY WEIGHTS

Animal #	Initial Weight (kg)	Final Weight (kg)
432	2.62	2.58
438	2.68	2.73
442	2.24	2.32
446	2.53	2.59
447	2.47	2.47
451	2.85	2.79
Mean	2.57	2.58
+SD	0.21	0.17

Table 3: INDIVIDUAL EYE IRRITATION SCORES

Animal # 432		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	1	1	0	0
	B. Chemosis	0	0	0	0
	C. Discharge	0	0	0	0
	Subtotal (A+B+C)x2	2	2	0	0

 

Animal # 438		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	1	1	0	0
	B. Chemosis	0	0	0	0
	C. Discharge	1	1	0	0
	Subtotal (A+B+C)x2	4	4	0	0

 

Animal # 442		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	1	1	0	0
	B. Chemosis	0	0	0	0
	C. Discharge	1	0	0	0
	Subtotal (A+B+C)x2	4	2	0	0

 

Animal # 446		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	0	0	0	0
	B. Chemosis	0	0	0	0
	C. Discharge	0	0	0	0
	Subtotal (A+B+C)x2	0	0	0	0

 

Animal # 447		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	1	1	1	0
	B. Chemosis	0	0	0	0
	C. Discharge	0	0	0	0
	Subtotal (A+B+C)x2	2	2	2	0

 

Animal # 451		Observation (Hours)
		1	24	48	72
Cornea	A. Opacity	0	0	0	0
	B. Area	0	0	0	0
	Subtotal (AxBx5)	0	0	0	0
Iris	A. Iris	0	0	0	0
	Subtotal (Ax5)	0	0	0	0
Conjunctiva	A. Erythema	0	0	0	0
	B. Chemosis	0	0	0	0
	C. Discharge	0	0	0	0
	Subtotal (A+B+C)x2	0	0	0	0

 

Table 4: GROUP MEAN EYE IRRITATION SCORES

Animal #	Tissue	Observation (Hours)
		1	24	48	72
432	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	2	2	0	0
	Total	2	2	0	0
438	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	4	4	0	0
	Total	4	4	0	0
442	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	4	2	0	0
	Total	4	2	0	0
446	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	0	0	0	0
	Total	0	0	0	0
447	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	2	2	2	0
	Total	2	2	2	0
451	Cornea (AxBx5)	0	0	0	0
	Iris (Ax5)	0	0	0	0
	Conjunctiva (A + B + C)x2	0	0	0	0
	Total	0	0	0	0
Group Mean	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctiva	2.0	1.7	0.3	0
	Total	2.0	1.7	0.3	0

 

Cornea                A =density

                              B =area

Corneal score = (AxBx5)

Maximum score = 80

 

Iris                        Iris score = (Ax5)             

Maximum score = 10

 

Conjunctiva       A = erythema   

                               B =chemosis

                               C = discharge

                               Conjunctival score = (A+B+C) X 2

                               Maximum score = 20

 

Total Maximum Score = 110.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Conjunctival irritation was observed in 4/6 of the treated eyes at the 1 hour and 24 hour observation. The maximum group mean irritation score was 2.0/110.0 and occurred at the 1 hour observation. This score decreased to 0 by 72 hours post-instillation. Automate Yellow 8 Petroleum would be classified as Toxicity Category III for eye effects in accordance with EPA regulations. Based on the mean scores for the 24, 48 and 72 hr time-points for each animal, this material would not be classified under the GHS system of classification.

Executive summary:

Automate Yellow 8 Petroleum was instilled into the conjunctival sac of the right eye of six New Zealand Albino rabbits at a dose of 0.1 g. The eyes were observed and scored at 1, 24, 48 and 72 hours after treatment.

Conjunctival irritation was observed in 4/6 of the treated eyes at the 1 hour and 24 hour observations. Conjunctival irritation was observed in 1/6 animals at the 48 hour observation. The maximum group mean score was 2.0/110.0 and occurred at the 1 hour observation. This score decreased to 0 by 72 hours post-instillation.

Therefore, Automate Yellow 8 Petroleum would be classified as Toxicity Category III for eye effects in accordance with EPA regulations.

Based on the mean scores for the 24, 48 and 72 hr time-points for each animal, this material would not be classified under the GHS system of classification.