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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006/06/22-2006/11/20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
NA
GLP compliance:
yes (incl. certificate)
Type of study:
Buehler test
Justification for non-LLNA method:
This test was performed for a registration of the test material according to TSCA.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
Description, Identification and Housing
Young adult, Hartley-derived albino guinea pigs were received from Hilltop Lab Animals,
Inc., Scottdale, Pennsylvania. Upon receipt, plastic ear tags displaying unique
identification numbers were used to individually identify the animals. Cage cards displaying
at least the study number, animal number and sex were affixed to each cage. The animals were
housed individually in suspended stainless steel cages. All housing and care were based on the
standards recommended by the Guide for the Care and Use of Laboratory Animals

Environment
The animal room temperature and relative humidity ranges were 60-76°F (16-24°C) and 43-100%,
respectively. Environmental control equipment was monitored and adjusted as necessary to minimize
fluctuations in the animal room environment. Light timers were set to maintain a 12-hour light/12-hour
dark cycle and room ventilation was set to produce 10-15 air changes/hour. The room temperature and
relative humidity were recorded a minimum of once daily.

Food
PMI Certified Guinea Pig Chow #5026 (PMI Nutrition International) was provided ad libitum to the animals
throughout the study. The lot number and expiration date of each batch of diet used during the study
were recorded. The feed was analyzed and certified by the supplier for nutritional components and environmental
contaminants. Dietary limitations for various environmental contaminants, including heavy metals, pesticides,
polychlorinated biphenyls and total aflatoxin are set by the manufacturer.
Within these limits, contaminants which may have been present were not expected to compromise the purpose
of this study.

Water
Municipal tap water treated by reverse osmosis was available ad libitum throughout the study. The purified
water was supplied by an automatic watering system. Monitoring of the drinking water for contaminants is
conducted by the testing laboratory and the records are available for inspection. Within generally accepted
limits, contaminants which may have been present were not expected to compromise the purpose of this study.
The water meets the standards specified under the EPA National Drinking Water Regulations (40 CFR Part 141).

Acclimation
Upon receipt, the animals were removed randomly from the shipping cartons, examined by qualified personnel,
identified with plastic ear tags and then acclimated to the laboratory conditions for a minimum of five days.
The animals were observed daily for overt physical or behavioral abnormalities, general health/moribundity
and mortality.

Animal Selection
The animals chosen for study use were arbitrarily selected from healthy stock animals to avoid potential bias.
All animals received a detailed pretest observation prior to dosing. Only healthy animals were chosen for study
use. Females were nulliparous and nonpregnant. The male animals were approximately 7 weeks of age and weighed
393-468 grams prior to Induction 1 dosing. The female animals were approximately 9 weeks of age and weighed
413-484 grams prior to Induction 1 dosing.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: deinonized water, ethanol
Concentration / amount:
Induction: 25%
Challenge: 5%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: deinonized water, ethanol
Concentration / amount:
Induction: 25%
Challenge: 5%
No. of animals per dose:
10 male/10 female
Details on study design:
This study consisted of pilot groups, two topical range-finding groups, a test group, and a challenge control group.
A rechallenge control group was maintained on this study; however, the rechallenge procedure was not required since
the challenge results were definitive.

Preliminary Procedures
On the day prior to each dose administration, the guinea pigs had the hair removed with a small animal clipper with a No. 40 blade followed
by further close clipping with a Braur. Syncro Electric Razor (Model 7505). Care was taken to avoid abrading the skin.

Dosing
A 0.3 mL dose of the test article was placed on a 25 mm Hilltop chamber backed by adhesive tape (occlusive patch).
The chambers were then applied to the clipped surface as quickly as possible.
Following chamber application, the trunk of the animal was wrapped with elastic wrap which was secured with
adhesive tape to prevent removal of the chamber and the animal was returned to its cage.



Challenge controls:
yes
Positive control substance(s):
yes
Remarks:
Hexylcinnamaldehyde

Study design: in vivo (LLNA)

Statistics:
NA

Results and discussion

Positive control results:
Using 2-Hexylcinnamaldehyde (HCA) as a positive control, Charles River Laboratories, Spencerville, Ohio, has completed
a study during the past six months which provided historical control data for contact sensitization to this agent
utilizing the test system described herein (Modified Buehler Design). Following induction at 5% w/v HCA in ethanol
and challenge at levels of 1% and 2.5% w/v HCA in acetone, a contact sensitization response was observed, thereby
demonstrating the susceptibility of the test system to this sensitizing agent.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: historical data (pos. control)
Dose level:
2.5%
No. with + reactions:
12
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: historical data (pos. control). Dose level: 2.5%. No with. + reactions: 12.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: historical data (pos. control)
Dose level:
2.5%
No. with + reactions:
6
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: historical data (pos. control). Dose level: 2.5%. No with. + reactions: 6.0. Total no. in groups: 20.0.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: NA
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: NA

Any other information on results incl. tables

NA

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information
Conclusions:
N-butylpyridinium heptachlorodialuminate does not meet the criteria to be considered a contact sensitizer under the conditions tested. The results
of the 2-Hexylcinnamaldehyde historical control study demonstrated that a valid test was performed and indicated that the test design would detect potential contact sensitizers.
Executive summary:

Introduction

The dermal sensitization potential of N-butylpyridinium heptachlorodialuminate was evaluated in Hartley-derived albino guinea pigs.

Ten male and ten female guinea pigs were topically treated with 25% N-butylpyridinium heptachlorodialuminate w/w in deionized water once per week, for three consecutive weeks. Following a two-week rest period, a challenge was performed whereby the 20 test and 10 previously untreated (naive) challenge control guinea pigs were topically treated with 5% N-butylpyridinium heptachlorodialuminate w/w in deionized water. Challenge responses in the test animals were compared with those of the challenge control animals.

Results

N-butylpyridinium heptachlorodialuminate

Following challenge with 5% N-butylpyridinium heptachlorodialuminate w/w in deionized water, dermal reactions in the test and challenge control animals were limited to scores of 0 to +/- at the 24- and 48-hour scoring intervals. Group mean dermal scores were similar in the test and challenge control animals.

2-Hexylcinnamaldehyde

Using 2-Hexylcinnamaldehyde (HCA) as a positive control, Charles River Laboratories, Spencerville, Ohio, has completed a study during the past six months which provided historical control data for contact sensitization to this agent utilizing the test system described herein (Modified Buehler Design). Following induction at 5% w/v HCA in ethanol and challenge at levels of 1% and 2.5% w/v HCA in acetone, a contact sensitization response was observed, thereby demonstrating the susceptibility of the test system to this sensitizing agent.

Conclusion

N-butylpyridinium heptachlorodialuminate does not meet the criteria to be considered a contact sensitizer under the conditions tested. The results of the HCA historical. control study demonstrated that a valid test was performed and indicated that the test design would detect potential contact sensitizers.