5,12-dihydro-2,9-dimethylquino[2,3-b]acridine-7,14-dione

Endpoint summary

Description of key information

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.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

In vivo information already in existence and available to use.

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

Route:

intradermal and epicutaneous

Vehicle:

other: Freund's Adjuvant, semi-liquid paraffin and petrolatum

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)

Route:

epicutaneous, occlusive

Vehicle:

petrolatum

Concentration / amount:

Dermal challenge on day 22:
5% in petrolatum (0.5 g applied)

Adequacy of challenge:

not specified

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

Key result

Group:

positive control

Remarks on result:

other: Positive ontrols were not included in the study but regularly tested in separate studies at the time of performance of this study

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

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.

Key result

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

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.

Key result

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.

Key result

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.

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.

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.

Reference 2

Endpoint:

skin sensitisation: in chemico

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available

Reference 3

Endpoint:

skin sensitisation: in vitro

Data waiving:

study technically not feasible

Justification for data waiving:

an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

No classification

Testing for sensitizing properties of the test item was performed according to the method of MAGNUSSON & KLIGMAN. 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.