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

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

6-4-1993 to 7-12-1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: EPA 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:

according to guideline

Guideline:

EPA OPP 81-6 (Skin Sensitisation)

Deviations:

not specified

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

study performed prrior to the requirement to perform the LLNA as the first choice method for skin sensitising potential.

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. Records of the test article stability, purity, source, and other data required by Federal Regulations will be maintained by the Sponsor.

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male

Details on test animals and environmental conditions:

Young adult male Hartley albino guinea pigs were used for this study and weighed between 230 and 298 grams at study initiation. The guinea pigs were obtained from Oak Hill, Holland, Michigan.
The guinea pigs used for this study were identified by ear tags and were individually housed in stainless steel cages in a temperature, humidity, and light-controlled room (room 15A). The animals were maintained according to the recommendations contained in DHHS Publications No. 86-23 (NIH): Revised 1985, "Guide for the Care and Use of Laboratory Animals." They were conditioned for 5 days prior to initiation of the study. Purina Guinea Pig chow and water were available ad libitum. The animals were selected randomly from the acclimated colony and assigned to the test groups. All animals used for this study appeared to be in good health at study initiation.

Justification for Selection of Test System: The albino guinea pig has been used extensively as an animal model for dermal sensitization studies.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Induction 2

Concentration / amount:

0.5 g of undiluted test material.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Challenge 2

Concentration / amount:

0.5 g of undiluted test material.

No. of animals per dose:

The test material was administered topically to ten guinea pigs.

Details on study design:

The test material was administered undiluted for all induction and challenge doses. Data from a primary skin irritation study indicated that the test article administered undiluted caused no skin irritation. The test material was administered topically to ten guinea pigs according to a modification of the method of Buehler (Buehler, E.V., Delayed contact hypersensitivity in the guinea pig. Arch. Dermatol. 91 :171-175, 1965).
An area of approximately 2 x 2 cm was shaved (electric clippers) on the left trunk of each animal on the first day of dosing. 0.5 grams of the test material was flattened into a wafer, and placed into a Hilltop Chamber with the pad removed. The test site was lubricated with 0.2 ml of mineral oil since the test material acted as a depilatory upon removal. The chamber was then applied to the test site, overwrapped (around the entire trunk) with Micropore, and the animal returned to its cage. The animal was unwrapped and the chamber removed after a 24 hour (initial dose) or six hour (nine succeeding induction doses) contact period. Animals were treated at least three times weekly (with at least one day intervening between treatments) for a total of ten treatments. Animals were reshaven when necessary prior to the application of a chamber. Two weeks after the final induction dose, each animal received a topical challenge dose (24 hr. contact) on a 2 x 2 cm shaved area located on the right trunk. Animals were scored for irritation 24 and 48 hours after each application.

Challenge controls:

Using the same regimen, six guinea pigs received 0.5 ml of a 0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol, to serve as a positive control. Two negative control animals were also challenged using the test material (24 hour contact period). The remaining two negative controls remained untreated.

Positive control substance(s):

yes

Remarks:

0.5 ml of a 0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol

Results and discussion

Positive control results:

The initial dose of 1-chloro 2,4 dinitrobenzene produced no positive irritation scores in any of the guinea pigs. The ten dose average irritation score was 0.39 for 1-chloro 2,4 dinitrobenzene. The average score for the challenge dose of 1- chloro 2,4 dinitrobenzene was 1.08. The challenge dose of 1-chloro 2,4 dinitrobenzene resulted in a positive reaction in 6/6 guinea pigs characterized by a grade 1 for erythema and/or edema.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No data..

Results 2

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No data..

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

2

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 2.0. Clinical observations: No data..

Results 4

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

2

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 2.0. Clinical observations: No data..

Results 5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol

No. with + reactions:

6

Total no. in group:

6

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: No data..

Results 6

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

positive control

Dose level:

0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol

No. with + reactions:

6

Total no. in group:

6

Clinical observations:

No data.

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.1 % solution of 1-chloro 2,4 dinitrobenzene in 80% aqueous ethanol. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: No data..

Any other information on results incl. tables

Appendix 1: INDIVIDUAL AND GROUP MEAN BODY WEIGHTS

 

Test Group (Automate Yellow 8 Petroleum)

Animal #	Initial Weight (g)	7-Day Weight (g)	14-Day Weight (g)	21-Day Weight (g)	Final Weight (g)
342	268	317	397	472	635
343	298	387	484	568	742
345	291	364	428	531	679
357	230	286	336	394	540
358	250	312	376	450	626
365	286	315	383	450	598
370	283	330	405	468	605
375	272	310	388	481	624
383	257	305	370	439	573
386	277	320	392	458	595
Mean	271	325	396	471	622
+/- S.D.	21	30	39	48	56

 

Positive Control Group (1-Chloro-2,4-Dinitrobenzene)

Animal #	Initial Weight (g)	7-Day Weight (g)	14-Day Weight (g)	21-Day Weight (g)	Final Weight (g)
337	290	363	456	541	709
340	281	323	392	460	638
349	305	390	465	556	713
352	241	327	394	478	630
381	274	316	374	423	506
389	265	344	411	481	552
Mean	276	344	415	490	625
+/- S.D.	22	28	37	50	83

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on comparisons of the initial test dose response to the challenge test dose and to the reactions elicited by the positive and negative controls, Automate Yellow 8 Petroleum would not be considered a dermal sensitizing agent.

Executive summary:

Automate Yellow 8 Petroleum (0.5 g) was placed directly into a Hilltop Chamber and applied to the shaved left trunk of ten albino guinea pigs. The chamber was then overwrapped (around the entire trunk) with Micropore tape and further secured with a wrap of surgical tape The animals were unwrapped after a 24 hour exposure period (initial dose). Nine succeeding induction doses were unwrapped after a 6 hour exposure period. This procedure was repeated three times weekly (with at least one day intervening between treatments) for a total of ten applications. Two weeks after the final application the animals received a topical challenge dose (24 hour contact) at a naive site located on the right trunk. Animals were scored for irritation 24 and 48 hours after each application.

Using the same regimen, six guinea pigs received 0.5 ml of a 0.1 % solution of 1-chloro 2,4 dinitrobenzene in (80% aqueous ethanol) to serve as a positive control. Four guinea pigs were observed as a negative control. Two negative control animals were also challenged using the test material.

The initial dose of Automate Yellow 8 Petroleum or 1-chloro 2,4 dinitrobenzene produced no positive irritation scores in any of the guinea pigs. The ten dose average irritation score was 0 for Automate Yellow 8 Petroleum. The ten dose average irritation score for 1-chloro 2,4 dinitrobenzene was 0.39.

Average scores for the challenge dose were 0 for Automate Yellow 8 Petroleum and 0 for the challenged negative controls. The average score for the challenge dose of 1-chloro 2,4 dinitrobenzene was 1.08.

The challenge dose of Automate Yellow 8 Petroleum, and two negative control animals challenged with Automate Yellow 8 Petroleum, did not elicit a positive response. The challenge dose of 1-chloro 2,4 dinitrobenzene resulted in a positive response in 6/6 guinea pigs,

characterized by a grade 1 for erythema and/or edema.

Based on comparisons of the initial test dose response to the challenge test dose and to the reactions elicited by the positive and negative controls, Automate Yellow 8 Petroleum would not be considered a dermal sensitizing agent.