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Description of key information

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 1998-07-13 to 1998-10-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study performed under GLP without deviations
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:
Buehler test
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Gartenstrasse 27, Box 1161, D-33176 Borchen.
- Age at study initiation: 6 weeks
- Weight at study initiation: 329 g to 419 g
- Housing: Makrolon cages type III (23 cm x 39 cm x 18 cm) with wire mesh lids, single caging.
- Diet (e.g. ad libitum): Altromin Standard Diet No. 3022, ad libitum, offered in stainless steel containers.
- Water (e.g. ad libitum): Tap water, acidified with HCl to pH = 3, offered in Makrolon bottles with stainless steel canules, ad libitum.
- Acclimation period: Approximately 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 60
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
Deionised
Concentration / amount:
Preliminary study: 40 %, 20 %, 5 % and 1 % in deionised water
Main study: 40% in deionised water
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
Deionised
Concentration / amount:
Preliminary study: 40 %, 20 %, 5 % and 1 % in deionised water
Main study: 40% in deionised water
No. of animals per dose:
Preliminary study: 3 females
Main study: 20 females for test substance group, 10 females for control group
Details on study design:
RANGE FINDING TESTS:
All concentrations were derived from a preliminary study with 3 female guinea pigs (epicutaneous exposure, exposure time 6 hours, same mode of administration as in the definitive study). The test substance concentrations were: 40 %, 20 %, 5 % and 1 % in deionised water. 40 % of the test substance in deionised water was the highest technically feasible concentration. No adverse skin reactions were observed at any of these concentrations in any animal. Therefore it was decided to use 40 % for the epicutaneous induction and for the challenge exposure.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6 h
- Test groups: test substance in deionised water
- Control group: deionised water
- Site: left flanks
- Frequency of applications: day 0, 7 and 15
- Duration: 0-21 day
- Concentrations: same throughout (40% (w/v) in deionised water)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 28
- Exposure period: 6 hours
- Test groups: posterior right flank: test substance (40 % (w/v) in deionised water), anterior right flank: deionised water
- Control group: posterior right flank: test substance (40 % (w/v) in deionised water), anterior right flank: deionised water
- Site: right flank
- Concentrations: same throughout (40% (w/v) in deionised water)
- Evaluation (hr after challenge): 24, 48
Positive control substance(s):
yes
Remarks:
HEXYL CINNAMIC ALDEHYDE
Statistics:
After challenge exposure, the number of animals with skin reactions on the test substance treated areas was recorded. A skin reaction was regarded as "positive", if the test substance treated area gained a higher score than the control area of the same animal. The potential of the test substance to cause skin sensitisation was established by subtracting the percentage of animals exhibiting an inflammatory reaction in the control group from the percentage of positively reacting test substance group animals. Differences in the body weights between the test substance group and the control group on Days 0 and 30 were compared by the t-test (two sided), p = 0.05.
Positive control results:
17/20 animals, i.e. 85 % of the animals of the positive control group, were regarded as sensitised. The results prove the sensitivity of the strain of animals used and the reliability of the experimental technique.
Reading:
other: any reading time
Group:
negative control
Dose level:
0.5 mL of 40% (w/v) solution
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
None
Remarks on result:
other: Reading: other: any reading time. Group: negative control. Dose level: 0.5 mL of 40% (w/v) solution . No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: None.
Reading:
other: any reading time
Group:
test chemical
Dose level:
0.5 mL of 40% (w/v) solution
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
None
Remarks on result:
other: Reading: other: any reading time. Group: test group. Dose level: 0.5 mL of 40% (w/v) solution. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: None.

Skin reactions after induction exposures

Group K1 (negative control): No adverse skin reactions in any animal at any time.

Group B (test substance): Severe erythema and/or oedema after the third induction exposure in 1/20 animals.

Skin reactions after challenge exposure

These were the reactions of interest for the grading of the allergenic potency of the test substance.

Group Kl (negative control):

Vehicle area: no positive skin reaction in any animal at any reading time.

Test substance area: no positive skin reaction in any animal at any reading time.

No animal had a "positive skin reaction".

Group B (test substance):

Vehicle area: no positive skin reaction in any animal at any reading time.

Test substance area: no positive skin reaction in any animal at any reading time.

No animal had a "positive skin reaction".

Mortality

All animals survived till the end of the study.

Body weight

There were no significant differences in mean body weights between the test substance group and the control group on Days 0 and 30.

General observations

Immediately after the beginning of all exposures (inductions, challenges) the motor activities of all animals were increased. This is due to the dressings which restrict the freedom of movement. Soon afterwards the behaviour was normal again. Except for the observations described above no abnormal behaviour or clinical signs of the animals were detected.

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Classification according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and CLP (Regulation (EC) No 1272/2008) is not required.
Executive summary:

The "Buehler test" of E.V. Buehler was performed to reveal a possible sensitising potential of Guanidine carbonate.

Twenty female guinea pigs were used as a test substance group and another 10 females were used as a negative control group. There were three epicutaneous induction exposures and one epicutaneous challenge exposure. The concentration of the test substance was 40 % (w/v) in deionised water for all three induction exposures and for the challenge exposure. The areas of administration were covered occlusively for 6 hours. Investigations performed were in conformance with the OECD TG 406.

All animals survived till the end of the study. Possible skin reactions excluded, no other adverse effects were noted. The test substance did not stain the skin of the guinea pigs and scoring was not impeded by skin dyeing. The application sites of all control animals were normal at each time. In the test substance group, erythema and/or oedema were noted in 1/20 animals after the third induction exposure.

In the negative control group, the control areas of all animals were normal 24 and 48 hours after the end of the challenge exposure.

All test substance treated areas of all animals of the negative control group and of the test substance group were also unaffected. No animal was regarded as irritated or sensitised.

Classification according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and CLP (Regulation (EC) No 1272/2008) is not required.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Skin sensitisation

One fully reliable study is available (Kocsis 1998) conducted according to OECD TG 406 (Bühler Test) and GLP. Additionally, a second study (Kocsis 1998) conducted according to OECD TG 406 (GPMT) with minor limitations is available.

In the Bühler assay, 20 female guinea pigs were treated with 40 % of the test substance in deionised water which was the highest technically feasible concentration both for the induction exposures as well as for the challenge exposure (occlusively for 6 hours). All animals survived until the end of the study and did not show signs of systemic toxicity. All test substance treated areas of all animals of the negative control group and of the test substance group were unaffected 24 and 48 hours post challenge treatment and accordingly no animal was regarded as irritated or sensitised.

In the study conducted according to the guinea pig maximisation test protocol 0.5 % of the test substance in physiological saline (w/v) for the intradermal induction, 40 % in deionised water (w/v) were used for the epicutaneous induction. 20 % in deionised water (w/v) were used for the first challenge exposure and 2 % in deionised water (w/v) were used for the second challenge exposure.

All animals survived until the end of the study and did not show signs of systemic toxicity. After the first challenge exposure, 80% of the test substance group animals had positive skin reactions, but also 60% of the control group animals. The result indicated an irritative effect of the test substance to the skin of guinea pigs at the concentration of 20 %. Therefore it was decided to perform a second challenge exposure with a lower test substance concentration (2 %). After the second challenge exposure, all test substance treated areas of all animals of the negative control group and of the test substance group were unaffected. No animal was regarded as irritated or sensitised.

It can be summarized that both in vivo tests show that Guanidine carbonate is not sensitising to the skin.

Respiratory sensitisation

No data on respiratory sensitisation is available.


Migrated from Short description of key information:
- skin sensitisation: not sensitising, 2 studies, OECD TG 406, Bühler Test and M&K Test
- respiratory sensitisation: no data available

Justification for selection of skin sensitisation endpoint:
A Bühler test and a guinea pig maximisation test are available. Both are performed under GLP and in accordance with OECD TG 406. The Bühler test is chosen as the key study because in the guinea pig maximisation test, the dose chosen for the challenge exposure was too high and the challenge had to be repeated with a lower dose.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

- respiratory sensitisation: No data available


Migrated from Short description of key information:
- respiratory sensitisation: No studies available

Justification for classification or non-classification

- skin sensitisation:

Based on the above stated assessment of the skin sensitisation potential of Guanidine carbonate (not sensitising to the skin of guinea pigs) the substance does not need to be classified according to CLP (Regulation (EC) No 1272/2008).

- respiratory sensitisation:

As no data on respiratory sensitisation is available for Guanidine carbonate a classification is not required according to CLP (Regulation (EC) No 1272/2008).

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