BRUGGOLITE® FF6

Endpoint summary

Description of key information

no adverse effect observed (not sensitising)

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:

December 1997 - July 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Planning and conduct of the study were based on the guidance provide by the following guidelines and directives: EEC Directive 96/54, L248, Annex IV C, Test B.6 (dated September 30, 1996) EEC Directive 93/21, L 110 A, Annex IV (dated May 04, 1993) OECD guidelines for the testing of chemicals (guideline 406 adopted July 17, 1992)

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Study was already available.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Sulzfeld
- Age at study initiation: approx. 28 days
- Weight at study initiation: 287 +/-10 g (n=15)
- Housing: Altromin Type S8/15, granulated soft wood bedding (Altromin, 32791 Lage/Lippe, Germany) Batch No. 051297 until 17/12/97, thereafter 181297
- Diet (ad libitum): Altromin 1322, standard diet for guinea pigs, Batch No: 040398/0725
- Water (ad libitum): tap water (municipal supply Bitterfeld) in Makrolon bottles, daily change
- Acclimation period: 6 days before the start of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23°C
- Humidity (%):30-60%
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): artificial ligth is set to give a cycle of 12 hours light and 12 hours dark

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

INDUCTION EXPOSURE:
Day 0: Induction by intradermal injection: Two pairs of intradermal injections of 0.1 ml each were administered to each animal.
Control group:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: Aqua pro injectione
Injection 3: the third injection was not necessary because the chosen vehicle was Aqua pro injectione
Dose group:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: 10% solution (w/v) of the test article in Aqua pro injectione
Injection 3: 10% emulsion (w/v) of the test article in 50:50 mixture (v/v) Aqua pro injectione and FCA

Day 7: Induction by dermal application
Control group: Filter papers (2x4 cm) were soaked in 0.1 g of white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.
Dose group:
Filter papers (2x4 cm) were soaked in 0.1 g of the 75:25 mixture (w/w) of test article and white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.

CHALLENGE EXPOSURE:
Day 21: Challenge by dermal application: each animal: 1.0 g sample of either vehicle or the 75:25 mixture (w/w) of test article and white vaseline

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

INDUCTION EXPOSURE:
Day 0: Induction by intradermal injection: Two pairs of intradermal injections of 0.1 ml each were administered to each animal.
Control group:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: Aqua pro injectione
Injection 3: the third injection was not necessary because the chosen vehicle was Aqua pro injectione
Dose group:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: 10% solution (w/v) of the test article in Aqua pro injectione
Injection 3: 10% emulsion (w/v) of the test article in 50:50 mixture (v/v) Aqua pro injectione and FCA

Day 7: Induction by dermal application
Control group: Filter papers (2x4 cm) were soaked in 0.1 g of white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.
Dose group:
Filter papers (2x4 cm) were soaked in 0.1 g of the 75:25 mixture (w/w) of test article and white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.

CHALLENGE EXPOSURE:
Day 21: Challenge by dermal application: each animal: 1.0 g sample of either vehicle or the 75:25 mixture (w/w) of test article and white vaseline

No. of animals per dose:

10 males (test group)
5 males (control group)

Details on study design:

RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
Induction by intradermal injection (Day 0)
- No. of exposures: 3
- Exposure period: -
- Test groups:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: 10% solution (w/v) of the test article in Aqua pro injectione
Injection 3: 10% emulsion (w/v) of the test article in 50:50 mixture (v/v) Aqua pro injectione and FCA

- Control group:
Injection 1: 50:50 mixture (v/v) of FCA and Aqua pro injectione
Injection 2: Aqua pro injectione
Injection 3: the third injection was not necessary because the chosen vehicle was Aqua pro injectione
- Site: shoulder region, each side of the midline in cranial to caudal sequence
- Frequency of applications:
- Duration:
- Concentrations:

Induction by dermal application (Day 7)
Dose group:
Filter papers (2x4 cm) were soaked in 0.1 g of the 75:25 mixture (w/w) of test article and white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.
Control group: Filter papers (2x4 cm) were soaked in 0.1 g of white vaseline. One paper each was then placed on the skin on both sides of the body and covered with an occlusive dressing for 48 hours.

B. CHALLENGE EXPOSURE
Challenge by dermal application (Day 21)
each animal: 1.0 g sample of either vehicle or the 75:25 mixture (w/w) of test article and white vaseline
- No. of exposures: each animal
- Day(s) of challenge: 21
- Exposure period: 24 hours
- Test groups: 1.0 g sample of either vehicle or the 75:25 mixture (w/w) of test article and white vaseline
- Control group: 1.0 g sample of either vehicle or the 75:25 mixture (w/w) of test article and white vaseline
- Site:dose group: right flank, controll group: left flank
- Concentrations: either vehicle or the 75:25 mixture (w/w) of test article and white vaseline
- Evaluation: 24 and 48 hours after patch removal (Days 23 and 24)

Challenge controls:

Guidelines recommend the use of guinea-pigs as the experimental rodent species for this type of study. The guinea-pig has been the mammal of choice for predictive sensitisation tests for several decades and a considerable data base is availabel from the literature.

Positive control substance(s):

yes

Remarks:

Freund's Complete Adjuvant (FCA)

Positive control results:

The positive control results demonstrate that the laboratory has the capability to identify positive dermal sensitizers.

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

vehicle (vaseline)

No. with + reactions:

0

Total no. in group:

15

Reading:

2nd reading

Hours after challenge:

72

Group:

negative control

Dose level:

vehicle (vaseline)

No. with + reactions:

0

Total no. in group:

15

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

test item in vaseline (75:25)

No. with + reactions:

0

Total no. in group:

10

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

test item in vaseline (75:25)

No. with + reactions:

0

Total no. in group:

10

Group:

positive control

Remarks on result:

other: no positive control was conducted

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

According to the classification system for skin reactions of the EEC Directive 93/21 Brüggolit FF6 has no skin sensitisation effect.

Executive summary:

The skin sensitisation potential of the test article was investigated in the Guinea-pig Maximisation Test with the Dunkin Hartley albino strain, whose sensitivity to benzocaine has been demonstrated.

In the course of testing, no clinical signs were observed and the body weight gain of animals was not significantly influenced. The choice of doses was based on the results ofthe pilot study.

The animals showed a very homogeneous reaction to the application of the test substance.

The intradermal injection of the 10 % solution of the test article alone and in combination with sensitisation potentiating FCA lead to slight erythema and oedema.

The same pattern was seen at injection site 1 (FCA/water mixture ) in the control animals. Topical induction was attempted with the 75:25 mixture of test article with white vaseline on day 7.

After removal of the occlusive dressing on day 9 not any skin irritation was recorded in the animals.

After challenge with the 75:25 preparation in white vaseline neither the control nor the dose group animals showed any skin effects. Oedema formation was excluded by measurement of skin-fold thickness.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

The skin sensitisation of the test item was tested according to OECD 406 with a 10% solution respectively a 75:25 mixture of the test substance solution with vaseline.

No skin irritation was observed, not any sensitisation reaction was registered and the skin fold thickness did not increase.

Migrated from Short description of key information:
skin sensitisation: No skin irritation (dermal induction); no sensitisation reaction; no increase in skin fold thickness

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

In the occupational medical assessment it was stated that a long-term exposure of the substance does not induce any health effects on affected employees. Neither reductions of the pulmonary function appeared nor any indication for an inhalative sensitization due to the inhalation of dust was given. This also applies for effects on the respiratory tract and the mucous membranes.

Migrated from Short description of key information:
No signs for reductions of the pulmonary function, no indication for an inhalative sensitization due to the inhalation of product dust. No indication of a development of malignant neoplasia. No indication of specific target-organic toxicity . No indication of effects on the respiratory tract and the
mucous membranes.

Justification for classification or non-classification

The test item needs not to be classified according to directive EC No. 1272/2008 due to the test results.