Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 July 1995 to 4 September 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with OECD Guideline 406 and EU Method B.6

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright-White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
-Strain specifics: Hoe: DHPK (SPFLac)
- Source: Hoechst AG, Kastengrund, SPF breeding colony
- Weight at study initiation: mean 385 g (range: 352-431 g)
- Housing: in groups of five animals in Makrolon type 4 cages in fully air-conditioned rooms
- Diet (e.g. ad libitum): standard diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: not necessary (breeding at identical conditions)
- Randomisation: randomisation scheme 95.0421

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): fully air-conditioned
- Photoperiod (hrs dark / hrs light):12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Freund's Adjuvant, semi-liquid paraffin and petrolatum (see below)
Concentration / amount:
Intradermal induction on day 1:
1% in semi-liquid paraffin, injection site 2
1% in 50% Freund's Adjuvant, injection site 3
(50% Freund's Adjuvant injected without test item at injection site 1)
Dermal induction on day 8:
25% in petrolatum (0.5 g applied)
Dermal challenge on day 22:
5% in petrolatum (0.5 g applied)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Freund's Adjuvant, semi-liquid paraffin and petrolatum (see below)
Concentration / amount:
Intradermal induction on day 1:
1% in semi-liquid paraffin, injection site 2
1% in 50% Freund's Adjuvant, injection site 3
(50% Freund's Adjuvant injected without test item at injection site 1)
Dermal induction on day 8:
25% in petrolatum (0.5 g applied)
Dermal challenge on day 22:
5% in petrolatum (0.5 g applied)
No. of animals per dose:
10 animals in the treatment group
5 animals in the control group
(in case of a questionable result additional animals will be tested to give a total of 20 test and 10 control animals)

6 animals in the group for determination of a primary non-irritant concentration
3 animals in the group for determination of tolerance of intradermal injections
5 animals in the escort group
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 intradermal injections and 1 dermal application
- Exposure period: intradermal injections on day 1 and dermal application on day 8 (occlusive, bandage removed after 48 h)
- Test groups: 3 sites (50% Freund's Adjuvant only, test substance in semi-liquid paraffin and test substance in 50% Freund's Adjuvant) per injection,test substance in petrolatum for dermal application
- Control and escort group: 3 sites (50% Freund's Adjuvant, semi-liquid paraffin and 50% Freund's Adjuvant) per injection, petrolatum for dermal application
- Site: dorsal area measuring 2 x 4 cm in the vicinity of the shoulders for intradermal injections, dermal application covered the same area where the intradermal injections had been placed
- Frequency of applications and duration: see above
- Concentrations: see above

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: occlusive for 24 h (removal of bandage on day 23)
- Test groups: test substance
- Control group: test substance
- Site: L flank
- Concentrations: see above
- Evaluation (hr after challenge): 48 h and 72 h
Challenge controls:
5 animals treated with the test substance (5% in petrolatum, 0.5 g applied)
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs of intoxication
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no clinical signs of intoxication.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs of intoxication
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no clinical signs of intoxication.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 5.0.

Any other information on results incl. tables

No effects on body weight gains or clinical signs were observed. The intradermal injection with Freund's adjuvant (with and without test substance) caused severe edema as well as indurations and encrustations. The application sites treated with Freund's adjuvant without the test substance showed severe erythema; the sites treated with the test substance showed slight edema. The injection sites treated with the test substance in Freund's adjuvant and in the vehicle were discolored pink, therefore, erythema formation was not assessable. Intradermal injections of the vehicle alone exhibited very slight erythema and edema. Due to these strong irritation reactions of the skin, treatment with 10% sodium dodecylsulfate was not done on day 7. Irritation readings during induction phase

Dermal Induction treatment (day 15)

Escort group animals 16 17 18 19 20

1. reading (48 hours)

Erythema 0 0 0 0 0  

Edema 0 0 0 0 0

Pink discoloration x x x x x  

2. reading (72 hours)

Erythema 0 0 0 0 0  

Edema 0 0 0 0 0

Pink discoloration x x x x x  

After the removal of the patches at day 10, severe erythema (only in the control group) and oedema, indurated and encrusted skin as well as necrosis were observed at the sites previously treated with Freund's Adjuvant. Additionally, the treated skin was disoloured pink in the animals of the treatment group, therefore erythema formation was not assessable. The application sites treated with the test substance in the vehicle showed slight oedema and indurations. No signs of irritation were observed at the application sites treated with the vehicle alone.

Skin readings after challenge treatment (day 29)

Control animals No. 1 2 3 4 5

Treatment animals No. 6 7 8 9 10 11 12 13 14 15

1. reading (48 hours)

Control animals

Erythema 0 0 0 0 0  

Edema 0 0 0 0 0

Pink discoloration x x x x x  

Treatment animals

Erythema 0 0 0 0 0 0 0 0 0 0

Edema 0 0 0 0 0 0 0 0 0 0

Pink discoloration x x x x x x x x x x

2. reading (72 hours)

Control animals

Erythema 0 0 0 0 0  

Edema 0 0 0 0 0

Pink discoloration x x x x x  

Treatment animals

Erythema 0 0 0 0 0 0 0 0 0 0

Edema 0 0 0 0 0 0 0 0 0 0

Pink discoloration x x x x x x x x x x

Although the skin surface was discolored light pink, possible erythema formation was assessable. No signs of irritation were observed in the control and the treatment group 24 and 48 hours after removal of the occlusive bandage.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
In a guinea pig maximisation test, the test item caused no skin reaction in the ten animals of the treatment group or in the five control animals. Based on the results of this study the test item showed no evidence for sensitising properties. The test substance is considered relevant and adequate to assess skin sensitisation properties of Pigment Red 122.
Executive summary:

Testing for sensitizing properties of the test item was performed in female Guinea pigs according to the method of MAGNUSSON & KLIGMAN. Intradermal induction was performed using 1 % test item in semi-liquid paraffin. Dermal induction was carried out with 25 % test item in petrolatum. Challenge treatment were carried out with 5 % test item in petrolatum. The challenge treatment caused no skin reaction in the ten animals of the treatment group or in the five control animals. Based on the results of this study the test item showed no evidence for sensitizing properties.