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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:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Identification: ALkYL (CI3-C15)·GLYCIDYL ETHER
Description: liquid
Batch Number: DC 1295.3

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Himalayan spotted
Sex:
male/female
Details on test animals and environmental conditions:
Test system
Rationale
Source
Ibm: GOHI: SPF-quality guinea pigs
(synonym: Himalayan spotted)
Recognized by the international guidelines
as the recommended test system, (e.g. DECO, EEC).
8Rl. Biological Research Laboratories· ltd.
W51ferstrasse 4
CH·4414 Fullinsdorf
18 mal as (3 malas used for pretest)
18 females (3 females used for pretest)
males : 7 weeks
females: B weeks
Randomization
Acclimatization
males: 332· 358 9
females: 323· 342 g
By unique cage number and corresponding ear tags.
Randomly selected at time of delivery.
One week under test conditions after
veterinary e~amination.
Test system: Ibm: GOHI: SPF-quality guinea pigs (synonym: Himalayan spotted)
Rationale: Recognized by the international guidelines as the recommended test system, (e.g. DECO, EEC).
Source: BRl. Biological Research Laboratories· ltd., Woelferstrasse 4 CH·4414 Fullinsdorf
Number of animals: 18 males (3 males used for pretest); 18 females (3 females used for pretest)
Age at Acclimatization start: males: 7 weeks; females: 8 weeks
Body Weight at Acclimatization Start: males: 332- 358g.; females: 323-342 g
Identification: By unique cage number and corresponding ear tags.
Randomization: Randomly selected at time of delivery.
Acclimatization: One week under test conditions after veterinary examination.
Standsrd Laboratory Conditions
Air-conditioned with 10·15 air changes per hour and hourly monitored environment
With a temperature of 22 + 3 degrees centigrade, a re.lative humidity between
40-70 %, 12 hours artificlal fluorescent lfght/12 hours dark, musiC during the
light period.
Accommodation
Individually in Makrolon type-3 cages with standard softwood bedding ("Ligno·
cel", Schi 11 AG, CH-4132 Muttenz).
Diet
Pelleted standard Kliba 342, Batches 59/90 and 60/90 guinea pig breedingl
maintenance diet ("Kliba", KlingentalmOhle AG, CH·4303 Kaiseraugst), ad libitum.
Results of analyses for contaminants are 1ncluded 1n th1s report.
Water
Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water.
Test system: Ibm: GOHI: SPF-quality guinea pigs (synonym: Himalayan spotted)

Rationale: Recognized by the international guidelines as the recommended test system, (e.g. DECO, EEC).

Source: BRl. Biological Research Laboratories• ltd., Woelferstrasse 4 CH•4414 Fullinsdorf

Number of animals: 18 males (3 males used for pretest); 18 females (3 females used for pretest)

Age at Acclimatization start: males: 7 weeks; females: 8 weeks

Body Weight at Acclimatization Start: males: 332- 358g.; females: 323-342 g

Identification: By unique cage number and corresponding ear tags.

Randomization: Randomly selected at time of delivery.

Acclimatization: One week under test conditions after veterinary examination.

Standard Laboratory Conditions
Air-conditioned with 10-15 air changes per hour and hourly monitored environment with a temperature of 22 + 3 degrees centigrade, a relative humidity between 40-70 %, 12 hours artificial fluorescent light/12 hours dark, music during the light period.

Accommodation
Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schili AG, CH-4132 Muttenz).

Diet
Pelleted standard Kliba 342, Batches 59/90 and 60/90 guinea pig breeding/
maintenance diet ("Kliba", Klingentalmuhle AG, CH•4303 Kaiseraugst), ad libitum.
Results of analyses for contaminants are included in the final report.

Water
Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water.

Test system: Ibm: GOHI: SPF-quality guinea pigs (synonym: Himalayan spotted)

Rationale: Recognized by the international guidelines as the recommended test system, (e.g. DECO, EEC).

Source: BRL. Biological Research Laboratories• ltd., Woelferstrasse 4 CH•4414 Fullinsdorf

Number of animals: 18 males (3 males used for pretest); 18 females (3 females used for pretest)

Age at Acclimatization start: males: 7 weeks; females: 8 weeks

Body Weight at Acclimatization Start: males: 332- 358g.; females: 323-342 g

Identification: By unique cage number and corresponding ear tags.

Randomization: Randomly selected at time of delivery.

Acclimatization: One week under test conditions after veterinary examination.

Standard Laboratory Conditions
Air-conditioned with 10-15 air changes per hour and hourly monitored environment with a temperature of 22 +3 degrees centigrade, a relative humidity between 40-70 %, 12 hours artificial fluorescent light/12 hours dark, music during the light period.

Accommodation
Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schili AG, CH-4132 Muttenz).

Diet
Pelleted standard Kliba 342, Batches 59/90 and 60/90 guinea pig breeding/
maintenance diet ("Kliba", Klingentalmuhle AG, CH-4303 Kaiseraugst), ad libitum.
Results of analyses for contaminants are included in the final report.

Water
Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water.

TEST ANIMALS : Ibm: GOHI: SPF-quality guinea pigs (synonym: Himalayan spotted)
- Source: 8RL Biological Research Laboratories· ltd.; W51ferstrasse 4 CH·4414 Fullinsdorf
- Age at study initiation: males : 7 weeks; females: 8 weeks
- Weight at study initiation: males: 332· 358 9; females: 323· 342 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Ligno·cel", Schi 11 AG, CH-4132 Muttenz).
- Diet: Pelleted standard Kliba 342, Batches 59/90 and 60/90 guinea pig breeding/maintenance diet ("Kliba", KlingentalmOhle AG, CH·4303 Kaiseraugst), ad libitum. Results of analyses for contaminants are included in final report.
- Water: Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water. Results of analyses for contaminants are included in final report.
- Acclimation period: One week under test conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
Air-conditioned with 10-15 air changes per hour and hourly monitored environment with a temperature of 22 + 3 degrees centigrade, a relative humidity between 40-70 %, 12 hours; artificlal fluorescent light/12 hours dark, music during the light period.

IN-LIFE DATES: August 14th through November 9th, 1990

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: vaseline
Concentration / amount:
Induction:
Test group:
Intradermal
1) Freund's complete adjuvant 50:50 with physiological saline.
2) The test article, d1luted to 5 %with oleum arach1dis.
3) The test article at the concentration used in (2), emulsified in a 50:50
mixture of Freund's complete adjuvant and the vehicle used in (2).
Epicutaneous: 75% in vaseline

Control Group:
1) Freund's complete adjuvant 50:50 with physiolog1cal saline.
2) Vehicle used in (2) for test group_
3) Freund's complete adjuvant 50:50 with physiological sal1ne.

Challenge:
non-irritant concentration (25 % in vaseline)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline
Concentration / amount:
Induction:
Test group:
Intradermal
1) Freund's complete adjuvant 50:50 with physiological saline.
2) The test article, d1luted to 5 %with oleum arach1dis.
3) The test article at the concentration used in (2), emulsified in a 50:50
mixture of Freund's complete adjuvant and the vehicle used in (2).
Epicutaneous: 75% in vaseline

Control Group:
1) Freund's complete adjuvant 50:50 with physiolog1cal saline.
2) Vehicle used in (2) for test group_
3) Freund's complete adjuvant 50:50 with physiological sal1ne.

Challenge:
non-irritant concentration (25 % in vaseline)
No. of animals per dose:
5 males, 5 females for the control group and 10 males, 10 females for the test group.
One male, one female for the intracutaneous (I.C.) pretest
2 females for the epicutaneous (E.C.) application.
Details on study design:
RANGE FINDING TESTS:
Intradermal injections:
Intradermal injections (0.1 ml/site) were made into the clipped flank of two guinea-pigs at concentrations of 5, 3 and 1% of the test article in oleum
arachidis . The resulting dermal reactions were assessed 24 hours later.
Epidermlal applications:
Patches of filter paper (2 x 2 cm) were saturated with the undiluted test article and with concentrations of 75%, 50% and 25 % of the test article in
vaseline and applied to the clipped and shaved flanks of each of four guineapigs. The patches were covered by a strip of aluminum foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. This procedure ensured the intensive contact of the test article. The dressings were removed after an exposure period of 24 hours and the reaction sites were assessed for erythema and edema on a numerical basis of erythema and edema at 24 and 48 hours after removal of the dressings.

MAIN STUDY
Induction
Intradermal injections:
An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 ml/site) were
made at the border of a 4 x 6 cm area in the clipped reg10n as follows:
Test group:
1) Freund's complete adjuvant 50:50 with physiolog1cal saline.
2) The test article, d1luted to 5 %with oleum arach1dis.
3) The test article at the concentration used in (2), emulsified in a 50:50 mixture of Freund's complete adjuvant and the vehicle used in (2).
Control Group:
1) Freund's complete adjuvant 50:50 with physiolog1cal saline.
2) Vehicle used in (2) for test group_
3) Freund's complete adjuvant 50:50 with physiological sal1ne.

Epidermal applications:
One week after the injections, the scapular area (approximately 6 x 6 cm) was again clipped and shaved free of hair. A 2 x 4 cm patch of filter paper
was saturated with the test article (75% in vaseline) and placed over the injection sites of the test animals. The patch was coverad by aluminum foil
and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The dressings were left
in place for approximately 48 hours. The epidermal application procedure described ensured intensive contact of the test article.
The guinea-pigs of the control group were treated as described above with the omission of test article.
Reaction sites were assessed for erythema and edema 24 and 48 hours after removel of the dressing, using the numerical grading system described previously.

Challenge
The test and control guinea-pigs were challenged two weeks after the epidermal induction application.
Hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea-pig. Two patches j2 x 2cm) of filter paper ·were saturated with a) non-irritant concentration (25 % in vaseline) of the test article and b) With the vehicle only and applied to the (a) left flank and (b) right flank using the same method as for the epidermal application.
The dressings were removed approximately 24 hours later. The sites were assessed for erythema and edema 24 and 48 hours after removal of the dressing, using the numerical scoring system as described by Draize.
The control animals were treated in the same way as described above.

Re-challenge
A second challenge was performed two weeks after the first challenge.
The treatment procedure for the animals of the test group was similar as described for the first challenge with the exceptlon that the flanks of the
guinea-pigs and the vehicle used for the test article dilution were changed (a - vehicle; b - 25% test article in oleum arachides).
The control animals were treated With the vehicle alone on the left flank.
Reading of challenge reactions
The challenge site was evaluated 24 and 48 hours after removal of the patch. The readings were made under artificial fluorescent light (daylight spectrum). Redness constitutes the minimum criterion of an allergic reaction. Strongly sensitized antmals display a vivid redness, associated With indurated swelling. The reactions were scored on the basis of the Draize score described under 'Readings and Scoring".
Rating of allergenicity
Based upon the percentage of animals sensitized (24-hour reading). the test article was assigned to one of the following ftve grades of allergenic
potency, ranging from weak to extreme.

Sensitization Grade Classi flcation
Rate [%]
0 - 8 1 weak
9 - 28 2 mi ld
29 - 64 3 moderate
65 - 80 4 strong
8l - 100 5 extreme

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No toxic symptoms; no deaths
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No toxic symptoms; no deaths.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No toxic symptoms; no deaths
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No toxic symptoms; no deaths.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
No. with + reactions:
17
Total no. in group:
20
Clinical observations:
No toxic symptoms; no deaths
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. No with. + reactions: 17.0. Total no. in groups: 20.0. Clinical observations: No toxic symptoms; no deaths.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
11
Total no. in group:
20
Clinical observations:
No toxic symptoms; no deaths
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 11.0. Total no. in groups: 20.0. Clinical observations: No toxic symptoms; no deaths.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
The results of this study conclude extreme allergenic potency of the test article, ALKYL (C13-C15)-GLYCIDYLETHER. The results were interpreted
according to the rating of Magnusson and Kligman (1969).
According to EEC (European Economic Community) classificatiion criteria described in guidelines 63/461, September 16, 1983 and 67/548, May 1987, this article is considered to be a sensitizer.
Executive summary:

The purpose of this skin sensitization study was to assess the allergenic potential of ALKYL (C13-C15)-GLYCIDYLETHER when administered to the skin of albino guinea pigs. ALKYL (CI3-CI5)-GLYCIOYLETHER was administered according to the Maximization-Test of B. Magnusson and A.M. Kligman (1969). Ten animals (5 males, 5 females) were used as control group and 20 animals (10 males, 10 females) were used as test group.

The study was conducted between August 14th and November and November 9th, 1990.