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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13. September - 19. October 1988
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report date:
1988

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
The modified Bühler test using 9 topical induction treatments was performed.
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
at that time, 1988, the LLNA according to OECD guideline 429 was not yet developed and the guinea pig the preferred test animal

Test material

Constituent 1
Chemical structure
Reference substance name:
reaction mass of: tert-alkyl(C12-C14)ammonium bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium bis[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium bis[1-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium [[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphthalenolato(2-)]-[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]]-chromate(1-) tert-alkyl(C12-C14)ammonium [[1-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-2-naphthalenolato(2-)]-[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]]-chromate(1-) tert-alkyl(C12-C14)ammonium ((1-(4-nitro-2-oxidophenylazo)-2-naphtholato)(1-(3-nitro-2-oxido-5-(1,1-dimethylpropyl)phenylazo)-2-naphtholato))chromate(1-)
EC Number:
938-781-3
Cas Number:
117527-94-3
Molecular formula:
C21H21N304.C16H11N304.Cr unspecified.C12-C14 chain unspec.
IUPAC Name:
reaction mass of: tert-alkyl(C12-C14)ammonium bis[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium bis[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium bis[1-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-2-naphthalenolato(2-)]-chromate(1-) tert-alkyl(C12-C14)ammonium [[1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphthalenolato(2-)]-[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]]-chromate(1-) tert-alkyl(C12-C14)ammonium [[1-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-2-naphthalenolato(2-)]-[1-[(2-hydroxy-5-nitrophenyl)azo]-2-naphthalenolato(2-)]]-chromate(1-) tert-alkyl(C12-C14)ammonium ((1-(4-nitro-2-oxidophenylazo)-2-naphtholato)(1-(3-nitro-2-oxido-5-(1,1-dimethylpropyl)phenylazo)-2-naphtholato))chromate(1-)
Details on test material:
- Lot/batch No.: 217342.89
- Appearance: Black powder
-Purity: not applicable (100% UVCB substance)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, FRG
- Age at study initiation: 2 months
- Weight at study initiation: 314 - 460 g
- Housing: metal cages wit wire-mesh floors/ 2 animals per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: a combined acclimation period of 13 days was observed (7 days for the primary irritation animals)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21 °C
- Humidity (%): 50-75

- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: To: 13 September - 19 October 1988

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: 1% methyl cellulose
Concentration / amount:
50% (w/w) in 1% methyl cellulose in milli-RO water
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: 1% methyl cellulose
Concentration / amount:
50% (w/w) in 1% methyl cellulose in milli-RO water
No. of animals per dose:
A total of 35 female animals were allocated into different groups: one experimental group of 20 animals and one group of 10 animals representing the control. The remaining five animals were used for primary irritation experiments.
Details on study design:
RANGE FINDING TESTS:
The primary irritation experiments included an epicutaneous treatment of one animal with a single concentration (50 % w/w) of the test substance in aqueous methyl cellulose 1% (mainly to assess any toxic effects) and an application with four guinea pigs of several concentrations of the test substance: 50%, 25%, 10% and 5% (w/w) in aqueous methyl cellulose 1% (to assess primary irritancy). The first animal was exposed to 0.05 ml of each concentration using Square chambers, both applications for 24 hours. The treated skin of the first animal showed no irritation. No signs of systemic toxicity were observed in any of the five treated animals.


MAIN STUDY
A. INDUCTION EXPOSURE
Experimental animals were exposed to the test substance, suspended to a concentration of 50% (w/w) in 1% methyl cellulose in milli-RO water and control animals to the vehicle. The patch consisted of a piece of Metalline (approx. 2x3 cm), attached to aluminium foil by means of a bit of petrolatum, that was mounted on Sleek tape. On day 0, the experimental animals were clipped in an area of the skin on the back just behind the right shoulder girdle. Thereafter the animals were wrapped in a "window dressing" which consisted of Coban elastic bandage with an opening of approximately 2x3 cm above the clipped area. Immediately thereafter, the area was exposed to dry ice for 5 seconds and a patch with 0.2 ml test substance was applied for 48 hours. On day 2 the same application was repeated. On day 4, the dressing was removed and the exposed area was clipped again. A new patch with 0.2 ml test substance was applied for 6 hours. Further induction exposures were carried out on the same site with 0.5 ml of the test substance, with an exposure period of 6 hours on days 7, 9, 11, 14, 16 and 18. The patch was covered with Coban elastic bandage. After each application the remaining test substance was removed from the skin with a tissue moistened with tap-water and if necessary, the exposed skin was clipped.
The control animals were treated in a similar manner to the experimental animals, on the left flank from day 4 onwards, with the exception that only the vehicle was applied. The skin reactions were read at the end of the induction phase.


B. CHALLENGE EXPOSURE
Ten days after the last induction exposure (day 28), the experimental and control animals were challenged to 50% (w/w) of the test substance in 1% methyl cellulose in milli-RO water and to the vehicle alone. The application was made to the clipped and shaved left flank of the experimental animals and on the right flank of the control animals using the same volume and the same patch as in the induction phase, and held in place with an elastic
Positive control substance(s):
yes
Remarks:
37% formaldehyde solution

Results and discussion

Positive control results:
A positive control experiment was carried out in May 1987. A sensitization rate of 50 per cent was obtained to the 1% formaldehyde
concentration

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50 % test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 % test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50% test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50% test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information