Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study performed according to the Test Guidelines B.6 of the EEC Directive 84/449/EEC and OECD Guideline for Testing of Chemicals n°406.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report date:
1989

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
A c. 700 g sample of the test substance was received from the sponsor on
September 19, 1988. It consisted of a fine, white powder, designated:
“AF—366”.

In vivo test system

Test animals

Species:
guinea pig
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
Young, SPF bred, albino guinea pigs (Bor:DHPW) were obtained from
F. Winkelmann, Institute for the Breeding of Laboratory Animals GmbH and
Co. KG, Borchen, F.R. Gerniany. During an acclimatization period of 8 days,
and throughout the test period, the animals were kept individually under
conventional conditions in suspended, stainless steel cages, fitted with
wire mesh floor and front in an animal room conditioned with respect to
temperature (21 ± 2°C), relative humidity (at least 40%), ventilation (at
least 10 air changes/hour) and lighting (12 hours light/12 hours dark
cycle). The animals were fed a pelleted, natural ingredient diet (Hope
Fans, Woerden, the Netherlands) and received unfluoridated tap water ad
libitum. Preliminary tests (to establish the concentrations of the test
substance to be used in the main study) were conducted on animals which had
already served as controls in a previous sensitization test, or with
animals never used before. The main study was carried Out with 15 males
(body weight 323—410 g) and 15 females (body weight 283—332 g) without
visible skin abnormalities. The animals ware received on February 14, 1989.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
maize oil
Concentration / amount:
induction treatment by intradermal injections of
Freund’s Complete Adjuvant (FCA), a 10% dilution (w/w) of the test
substance in maize oil, and a 10% dilution (w/w) of the test
substance in a mixture (1:1) of FCA and maize oil
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
maize oil
Concentration / amount:
induction treatment by intradermal injections of
Freund’s Complete Adjuvant (FCA), a 10% dilution (w/w) of the test
substance in maize oil, and a 10% dilution (w/w) of the test
substance in a mixture (1:1) of FCA and maize oil
No. of animals per dose:
20 test animals
and 10 controls
Details on study design:
The experiment was conducted essentially according to the guinea pig
maximization test as described by B. Magnusson and A.M. Kligman (1969 and
1970), and according to:
— EEC—Directive 84/449, Annex V, Part B: Methods for the determination of
toxicity, B.6. Acute toxicity, skin sensitization, dated September 1984,
— the QECD Guidelines for Testing of Chemicals, nr. 406, adopted 12 May
1981.
The study consisted of an induction treatment, followed by a resting period
of 14 days, which preceded the challenge treatment.
Preliminary observations were made to establish the concentrations of the
test substance to be used for intradermal injection and for topical
application in the main study.
Challenge controls:
The topical challenge was carried out two weeks after the topical induction
as follows:
An area of 5 x 5 cm on the left and right flank of each test and control
animal was clipped free from hair. Subsequently, cups were loaded with a
25% or a 50% dilution (w/w) of the test substance in vaseline or with
vaseline alone. The cups with the 25% and 50% test dilution were placed
separately on the shaved area of the left flank of each test and control
animal. The cup with vaseline alone was placed on the shaved area of the
right flank of each test and control animal. The cups were covered with
Leukopor bandage, and held in place by Tensoplast for 24 hours. Skin
readings were made at 24 and 48 hours after removal of the cups.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50% dilution (w/w) of “AF—366” in vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactions in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50% dilution (w/w) of “AF—366” in vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactions in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50% dilution (w/w) of “AF—366” in vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% dilution (w/w) of “AF—366” in vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25% dilution (w/w) of “AF—366” in vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% dilution (w/w) of “AF—366” in vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25% dilution (w/w) of “AF—366” in vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% dilution (w/w) of “AF—366” in vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
only vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: only vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
only vaseline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: only vaseline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: The challenge treatment with vaseline alone, did not induce skin reactionS in any of the test animals or controls. The chalienge treatment with the 25% and 50% test dilution in vaseline, did not induce skin reactions in any of the test animals or controls.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
On the basis of the resuits obtained under the conditions of this study, it
is concLuded that, according to the EEC-standards (as published in the
Official Journal of the European Communities, L257, Volume 26, 16 September
1983), the substance “AF-366” is not a sensitizer.