Registration Dossier

Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
Buehler method
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000.09.04 - 2000.10.20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

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
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
Buehler test
Justification for non-LLNA method:
In vivo data for MI3 skin sensitization to the guinea-pig was allready avalible prior initiating this registration dossier.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Cream powder (84% w/w)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: MI3, CHD NO. 0179/00-0665 Lot 2, Batch number: C202493
- Expiration date of the lot/batch: December 2000
- Purity 98.67 % (295 mg active sub/g crude sub)
- Purity test date: 2000.07.13

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: - 20 degrees celsius, in the dark.
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: up to 90 % w/v in water.


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Fresh prepared smooth brown paste (90 %w/v) using water on the day of exposure
- Final preparation of a solid: Freshly prepared smooth brown paste (90 %w/v) using water on the day of exposure

FORM AS APPLIED IN THE TEST (if different from that of starting material) freshly prepared paste (*90 %w/v) using water as vehicle.

:

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dunkin/Hartley, albino
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Microbiological status of animals, when known: NA
- Age at study initiation: 5 - 8 weeks old.
- Weight at study initiation: 351 - 427g
- Housing: groups of five animals in suspended metal cages with wire mesh floors
- Diet (e.g. ad libitum): ad libitum, vitamin C enriched guinea pig diet.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: five days
- Indication of any skin lesions: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24 degree celsius
- Humidity (%): 30 - 70 %
- Air changes (per hr): NA
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12hrs light
- IN-LIFE DATES: From: To: 2000.09.04 - 2000.10.20

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
MI3, 90% w/v in sterile water
Day(s)/duration:
on Days 1, 8 and 15.
Adequacy of induction:
highest technically applicable concentration used
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
MI3, 90% w/v in sterile water
Day(s)/duration:
Challange two weeks after the final induction and examinations after 24 and 48 hours.
Adequacy of challenge:
other: the maximum practical concentration that could be prepared and dosed topically and not given rise to irritating effects.
No. of animals per dose:
Control animals (n=10).
Test animals (n=20).
Preliminary investigations (n=4).
Details on study design:
RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: Day 1, Day 8 and Day 15.
- Test groups: MI3 exposure, 20 animals
- Control group: Vehicle (sterile water) exposure, 10 animals
- Site: Left shoulder region
- Frequency of applications: 3 times
- Duration: 6 hours
- Concentrations: 90% w/v in sterile water

B. CHALLENGE EXPOSURE
- No. of exposures: One
- Day(s) of challenge: Two weeks after final induction
- Exposure period: 6 hours
- Test groups: MI3 exposure, 20 animals
- Control group: Exposured as test group, 10 animals
- Site: The right flank
- Concentrations: 90% w/v in sterile water
- Evaluation (hr after challenge): 6 hours

OTHER:
Challenge controls:
yes as test animals challanged with MI3 90% v/v
Positive control substance(s):
yes
Remarks:
Hexyl cinnamic aldehyde (HCA) as 50% v/v in Alembicol D

Results and discussion

Positive control results:
Induction:
Slight to well-defined dermal reactions were observed for all test animals during the induction period and no dermal reactions were observed for the control animals.
Challenge:
The dermal reaction seen in all the ten test animals were more marked than those for controls, therefore the reactions seen in all of the test animals were considered to represent evidence of skin sensitization.
Conclusion:
In this study HCA produced evidence of skin sensitization in all of the ten animals, thus confirming the sensitivity and reliability of experimental technique to detect skin sensitization potential.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
sterile water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were observed
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
sterile water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were observed
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
MI3, 90% w/v in sterile water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No signs of ill health or toxicity were observed
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
MI3, 90% w/v in sterile water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No signs of ill health or toxicity were observed
Remarks on result:
no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The Study was conducted to determine the skin sensitization potential of MI3 using the guinea-pig. The study was designed in accordance with OECD guideline method 406 “modified Buehlers test”. The MI3 dose level was chosen to be 90% v/v from a preliminary study, and hexyl cinnamic aldehyde (HCA) was used as positive control. Twenty test and ten control female guinea-pigs received 3 induction applications with either MI3 (90% v/v) or water, respectively, and a challenge test two weeks after the final induction. There was no indicative of systemic toxicity or ill health for any of the guinea-pigs. In this study MI3 did not produce evidence of skin sensitization in any of the twenty test animals.
Executive summary:

The Study was conducted to determine the skin sensitization potential of MI3 using the guinea-pig. The study was designed in accordance with OECD guideline method 406 “modified Buehlers test” the commission directive 96/54/EC, Method B6, and the US EPA Health effects test Guidelines OPPTS 870.2600. The MI3 dose level was chosen to be 90% v/v from a preliminary study and hexyl cinnamic aldehyde (HCA) was used as positive control. Twenty test and ten control female guinea-pigs received 3 induction applications on the shoulder region at Day 1, 8 or 15 with either MI3 (90% v/v) or water, respectively. All animals were challenged test two weeks after the final induction and examined after 24 or 48 hours. There was no indicative of bodyweight loss, systemic toxicity or ill health for any of the guinea-pigs. All animals exposed to HCA showed positive evidence of skin sensitization after 48 hours. In the present study, MI3 did not produce evidence of skin sensitization in any of the twenty test animals.