PIGMENTADDITIV RL

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2 Mai to 5 Jun 1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD TG 406), according to GLP

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

(Council Directive 84/449/EEC, adopted25 April 1984)

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(adopted 12 Mai 1981)

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

OECD 429 was adopted as a new guideline for the testing of chemicals on April 24th, 2002

Species:

guinea pig

Strain:

other: Pirbright-White

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: HOECHST AG, Kastengrund, Germany
- Age at study initiation: not stated
- Weight at study initiation: mean 320 g (range 264-346 g)
- Housing: in groups of 5 per cage
- Diet (ad libitum): Altromin 3112 for guinea pigs (Altromin GmbH, Lage, Germany)
- Water (ad libitum): tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): not stated, fully air-conditioned
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

other: semi-liquid paraffin for intradermal injections, petrolatum for dermal treatment

Concentration / amount:

Determination of primary non-irritant concentration:0.25, 2.5, and 25 % in petrolatum
Determination of tolerance to intradermal injection: 0.2, 1 and 5 % in semi-liquid paraffin
Intradermal induction: 0.2 % in semi-liquid paraffin
Epicutaneous induction: 25 % in petrolatum
Epicutaneous challenge: 2.5 % in petrolatum

Route:

epicutaneous, occlusive

Vehicle:

other: semi-liquid paraffin for intradermal injections, petrolatum for dermal treatment

Concentration / amount:

Determination of primary non-irritant concentration:0.25, 2.5, and 25 % in petrolatum
Determination of tolerance to intradermal injection: 0.2, 1 and 5 % in semi-liquid paraffin
Intradermal induction: 0.2 % in semi-liquid paraffin
Epicutaneous induction: 25 % in petrolatum
Epicutaneous challenge: 2.5 % in petrolatum

No. of animals per dose:

10 in control group, 5 in exposure group, 5 in escort group

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (one intradermal on day 1, one epicutaneous on day 8 of study (48 hocclusive))
- Test groups:
intradermal:
2 x 0.1 mL per site; site 1: 50 % Freund's adjuvant; site 2: 0.2 % test item in semi-liquid paraffin; site 3: 0.2 % test item in 50% Freund's adjuvant
epicutaneous:
0.5 g of 25 % test item in petrolatum on a 2 x 4 cm cellulose patch under occlusive bandage

- Control and escort groups:
intradermal:
2 x 0.1 mL per site; site 1: 50 % Freund's adjuvant; site 2: semi-liquid paraffin; site 3: 50 % Freund's adjuvant
epicutaneous:
0.5 g petrolatum

- Site: dorsal area of 4 x 6 cm clipped, intradermal exposures within 2 x 4 cm; epicutaneous exposure to 2 x 4 cm


B. CHALLENGE EXPOSURE
- No. of exposures: 1 (epicutaneous)
- Day(s) of challenge: day 22
- Exposure period: 24 h occlusive
- Test groups: 0.5 g of 2.5 % test item in petrolatum
- Control group: 0.5 g of 2.5 % test item in petrolatum
- Site: 5 x 5 cm on left flank clipped, exposure to 2 x 2 cm under occlusice bandage
- Evaluation (hr after challenge): 24, 48, and 72 h

OTHER:
Escort group:
- induction identical to control group
- challenge identical to test and control groups on day 15-18 of study

Challenge controls:

Treatment of the animals with Freund's Adjuvant can lower the threshold value for primary irritation determined in preliminary tests. For this reason, the five animals in the escort group which had been treated with Freund's Adjuvant were treated with 25.0 % test item in petrolatum from day 14 to 18. One animal developed a slight erythema on the day after removal of the test substance. On the second day after removal of the test substance there were no signs of irritation in any of the animals. The skin of the animals was discoloured blue. On day 3 after removal 4/5 animals developed clearly bordered erythema and one animal a slight edema. Therefore a challenge concentration of 2.5% was selected.

Positive control substance(s):

no

Positive control results:

no positive control in study (not required by guideline version of 1981)

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

2.5%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

2.5%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

2.5%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Main test for sensitising properties 

The treated animals showed no clinical signs of intoxication throughout the study and body weights were within the normal range. The intradermal injections with Freund's adjuvant (with and without test substance) caused erythema and edema. About 3 days after application the injection sites partially showed eschar formation. Most of them developed necrosis about one week after application. Sites only treated with test item or vehicle showed slight erythema and edema which disappeared on the fifth day after application. Injection of the test item caused blue discoloration. After the removal of the induction patches at day 10 edema and necrosis were observed at the sites previously treated with Freund's adjuvant. Treatment of the injection sites with test item or vehicle alone did not cause irritation.

Challenge treatment No signs of irritation were observed in the control group and test group 24, 48, and 72 hours after removal of the occlusive bandage.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the test item was not sensitising

Executive summary:

A guinea pig maximisation test was performed to assess the cutaneous allergenic potential of the test item in accordance with OECD Guideline 406 and GLP. It was carried out on 15 female albino guinea pigs (5 test and 10 control animals). Intradermal induction was performed using 0.2 % test item in semi-liquid paraffin or Freund ’s adjuvant. The controls received semi-liquid paraffin or Freund’s adjuvant only. Dermal induction and challenge treatment were carried out with 25 % and 2.5 % test item in petrolatum, respectively. The treated animals showed no clinical signs of intoxication throughout the study. The intradermal injections with Freund's adjuvant (with and without test substance) caused erythema and edema and finally necrosis. The application sites treated with the test substance or the vehicle alone showed slight signs of irritation, which disappeared after 5 days. After the removal of the patch at day 10, necrosis and edema were observed at the sites previously treated with Freund's adjuvant. The body weight gains of the treated animals were not impaired. At the end of the challange treatment no signs of irritation were observed in the control group and test group 24, 48, and 72 hours after removal of the occlusive bandage.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

 

The test substance was not sensitising in a reliable guinea pig maximisation test, according to guideline and GLP (RL 1, Hoechst, 1989). For the intradermal induction a concentration of 0.2% test item in semi-liquid paraffin or Freund's Adjuvans was used. Epidermal induction was performed with 25% test item in petrolatum. Dermal challenge was performed with 2.5% test item in petrolatum. No skin sensitisation was observed after dermal challenge.

Migrated from Short description of key information:
The test substance was not sensitising in a reliable guinea pig maximisation test. There is no study on respiratory irritation available.

Justification for selection of skin sensitisation endpoint:
Guideline study (OECD TG 406), according to GLP

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

This information is not available.

Migrated from Short description of key information:
This information is not available.

Justification for classification or non-classification

Due to the negative finding in a reliable guinea pig maximisation test, no classification according to Regulation (EC) No 1272/2008 and Council Directive 67/548/EEC is required for skin sensitisation. There are no data on respiratory sensitisation.