Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-02-12 to 2013-03-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted 1992-07-17
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Potassium hexafluorotitanate (K2TiF6)
- Physical state: white powder
- Storage condition of test material: the test article was stored at room temperature and humidity.

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS - Hartley Albino guinea pigs
- Age at study initiation: 2 - 3.5 weeks
- Weight at study initiation: males: 277 - 345 g; females: 285 - 332 g
- Housing: individually housed in suspended wire cages
- Diet (ad libitum): Fresh PMI Guinea Pig Chow (Diet #5025)
- Water (ad libitum): water
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS - temperature controlled rooms
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Intradermal induction: 1% concentration
Topical induction: 75% concentration
Challenge: 10% concentration
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Intradermal induction: 1% concentration
Topical induction: 75% concentration
Challenge: 10% concentration
No. of animals per dose:
Test article group: 10 males / 10 females
Details on study design:
SITE PREPARATION
The day prior to the screen, intradermal induction, topical induction, or challenge, the sites (back or sides) were clipped free of hair with an electric clipper. Upon examination prior to the screen or intradermal induction, animals with skin irregularities or irritation were eliminated from the study.

RANGE FINDING TESTS:
The preliminary screen was conducted to determine the concentration for intradermal injection, which was not necrotic, ulcerogenic or toxic. For the topical application, the concentration of the test article must be well tolerated systemically and produce no more than mild to moderate skin irritation.
The test article was prepared in distilled water, to the following five concentrations: 1, 10, 25, 50 and 75%. Three guinea pigs were dosed intradermally with 0.1 ml of each of the following test article concentrations: 1, 10, 25, and 50%. Three other guinea pigs were dosed topically with 0.1 ml of each of the following test article concentrations: 10, 25, 50 and 75%. For the topical applications, 0.1 ml of the test article mixture was applied to 2 x 2 cm squares of Whatman’s #1 filter paper, which were applied to the skin. The patches, occluded with plastic and fastened with nonirritating tape, remained in place for 24 hours.
The intradermal sites were scored 24 hours and 3 days after dosing.
The topical sites were scored 24 and 48 hours after patch removal.
Erythema was evaluated using the Magnusson and Kligman grading scale. Additional signs were described.
The screen animals were weighed pretest and observed daily for mortality and toxicity.

Results:
Based on the results of the intradermal injections, a 1% concentration was chosen for intradermal induction.
Based on the results of the topical doses, a 75% concentration was chosen for topical induction.
Based on the results of the topical doses, a 10% concentration was chosen for the challenge.

MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal induction: Six intradermal injections were made on the 4 x 6 cm prepared site as follows:
- One site received 0.1 ml of 50% Freunds Complete Adjuvant (FCA) in distilled water on both sides of the midline
- One site received 0.1 ml of a 1% concentration of the test article mixture on both sides of the midline
- One site received 0.1 ml of a mixture containing equal parts of 50% FCA and a 1% concentration of the test article on both sides of the midline

Topical induction: because a 75% concentration of the test article did not produce irritation in the preliminary screen, the animals were pretreated with 0.5 ml of a 10% mixture of sodium dodecyl sulfate approximately 24 hours prior to topical induction and the sites remained unoccluded. At least two hours prior to topical induction, any residual sodium dodecyl sulfate was removed with distilled water. Seven days after intradermal induction, the guinea pigs in the test article group were dosed topically using 2 x 4 cm patches of Whatman’s #1 filter paper, saturated with 0.2 ml of the 75% concentration. The patches, occluded with plastic and fastened with adhesive tape, remained in place for 48 hours. The guinea pigs in the control group were dosed in the same manner with the vehicle control.

B. CHALLENGE EXPOSURE
Fourteen days after topical induction, the test and control animals were challenged topically using 2 x 2 cm patches of Whatman’s #1 filter paper, saturated with 0.1 ml of a 10% concentration of the test article one one side. The vehicle was applied topically on the other side. All sites were occluded with plastic and secured with non-irritating tape for 24 hours.

OBSERVATIONS:
- Intradermal injections: the treated site of each animal was examined and scored for dermal reactions at 24 and 48 hours after injection.
- Topical induction: the treated site of each animal was examined and scored at 48 hours after application of the saturated patch.
- Challenge: the challenge sites, both test article and control, of each animal were examined and scored at 24 and 48 hours after patch removal.

Erythema was evaluated using the Magnusson and Kligman grading scale. Additional signs were described.

Clinical signs: animals were observed once daily for mortality and toxicity
Body Weights: body weights were recorded pretest and at study termination.
Challenge controls:
Vehicle control group (100% distilled water): 5 males / 5 females
During challenge they received the test article as a 10% concentration.
Positive control substance(s):
yes
Remarks:
2-mercaptobenzothiazole 98% (5% in acetone for intradermal induction; 50% in acetone for topical induction; 25% in acetone for challenge)

Results and discussion

Positive control results:
Dermal observations:
- Test article group:
Intradermal induction: erythema was absent to intense with pale areas
Topical induction: erythema was absent to moderate
Challenge: erythema was absent to moderate on the test article site and absent on the vehicle control site

-Vehicle control group:
Intradermal induction: erythema was absent to moderate
Topical induction: erythema was absent to moderate
Challenge: erythema was absent in both the test article and vehicle control site

2-mercaptobenzothiazole 98% was considered to have a skin sensitising potential.

Body weight:
All animals gained weight during the study.

Systemic observations:
One male of the vehicle control group appeared emaciated. All other animals appeared normal.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10% concentration of test article
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
One male of the test article group had soiling of the anogenital area; all other animals appeared normal.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10% concentration of test article. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: One male of the test article group had soiling of the anogenital area; all other animals appeared normal..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10% concentration of test article
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: 10% concentration of test article. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100% distilled water
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: 100% distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100% distilled water
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: 100% distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10% concentration of test article
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
All animals appeared normal.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10% concentration of test article. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: All animals appeared normal..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10% concentration of test article
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10% concentration of test article. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100% distilled water
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100% distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100% distilled water
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100% distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25% concentration of test article
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
One male of the negative (vehicle) control group appeared emaciated. All other animals appeared normal.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 25% concentration of test article. No with. + reactions: 8.0. Total no. in groups: 20.0. Clinical observations: One male of the negative (vehicle) control group appeared emaciated. All other animals appeared normal..
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25% concentration of test article
No. with + reactions:
8
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 25% concentration of test article. No with. + reactions: 8.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
100% acetone
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 100% acetone. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
100% acetone
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 100% acetone. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: negative control in the separately conducted positive control study
Dose level:
25% concentration of test article
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group:
Reading:
2nd reading
Hours after challenge:
48
Group:
other: negative control in the separately conducted positive control study
Dose level:
25% concentration of test article
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group:
Reading:
1st reading
Hours after challenge:
24
Group:
other: negative control in the separately conducted positive control study
Dose level:
100% acetone
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group:
Reading:
2nd reading
Hours after challenge:
48
Group:
other: negative control in the separately conducted positive control study
Dose level:
100% acetone
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group:

Any other information on results incl. tables

Dermal observations:

- Test article group:

Intradermal induction: erythema was absent to intense

Topical induction: erythema was absent to moderate

Challenge: erythema was absent in both the test article site and the vehicle control site

-Vehicle control group:

Intradermal induction: erythema was absent to discrete

Topical induction: erythema was discrete to moderate

Challenge: erythema was absent in both the test article and vehicle control site

Body weights:

All animals gained weight during the study.

Systemic observations:

One male of the test article group had soiling of the anogenital area; all other animals appeared normal.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material is not considered to be a skin sensitiser. The test material produced a 0% (0/20) sensitisation rate.
According to 67/548/EC and subsequent regulations,dipotassium hexafluorotitanate is not classified as a skin sensitiser.
According to the EC Regulation No. 1272/2008 and subsequent regulations, dipotassium hexafluorotitanate is not classified as a skin sensitiser.