Aluminium triformate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 July 2009 to 3 December 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Fully GLP- and guideline compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

Principles of method if other than guideline:

Modifications according to Th. Maurer and R. Hess (1989): The maximization test for skin sensitization potential-updating the standard protocol and validation of a modified protocol. Fd Chem. Toxic. Vol. 27, No. 12, 807-811.

GLP compliance:

yes

Type of study:

Maurer optimisation test

Justification for non-LLNA method:

Maurer optimisation test was already available. See below for further justification:
Guinea pigs: animal of choice for predictive sensitisation tests
Discrimination of potential false-positive/negative results or to confirm previous data
Assessment of delayed contact hypersensitivity

Test material

Test material information

Constituent 1

Reference substance name:

Aluminium triformate

EC Number:

230-898-1

EC Name:

Aluminium triformate

Cas Number:

7360-53-4

Molecular formula:

CH2O2.1/3Al

IUPAC Name:

aluminium triformate

Details on test material:

- Name of test material (as cited in study report): Aluminium Triformate
- Trade name: Altriform CFD, Novaltan Al
- Hill formula: Al (HCOO)3.3H2O
- Physical state: White powder
- Composition of test material, percentage of components: 24.8 % Aluminiumoxide, 62 % formic acid
- Purity test date: 24 June 2009
- Lot/batch No.: 128110/004
- Expiration date of the lot/batch: not stated
- Stability under test conditions:
- Storage condition of test material: Room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, 88353 Kisslegg, Germany
- Age at study initiation: Approx. 5 - 7 weeks at the first application
- Weight at study initiation: Step 1: 351g - 382g, Step 2: 303g - 333g.
- Housing: Group caging in plastic containers (46 cm x 105 cm x 36 cm), partly shaded, 6 (control group) or 11 (test substance group) animals per container.
- Diet (e.g. ad libitum): Ssniff Ms-H (Guinea Pig Maintenance Diet V2233), including ascorbic acid (2400 mg/kg), ad libitum, offered in stainless steel containers. Analysis of the feed for ingredients and contaminants are performed randomly by ssniff Spezialdiäten GmbH, Ferdinand-Gabriel-Weg 16, 59494 Soest, Germany.
- Water (e.g. ad libitum): Tap water offered in Makrolon bottles with stainless steel canules ad libitum. Random samples of the water are analysed by the "AGES", A-1226 Vienna, to assure that the water fulfils the requirements for drinking water for humans.
- Acclimation period: Step 1: 12 days, Step 2: 5 days.


ENVIRONMENTAL CONDITIONS
- Temperature: Mean of 20.6°C
- Humidity: Step 1: Mean of 64.17 %, Step 2: Mean of 41.35 %.
- Air changes (per hr): 12
- Photoperiod: 12 hrs dark, 12 hrs light

IN-LIFE DATES: From: 11 August 2009 To: 20 November 2009

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

petrolatum

Remarks:

white peotrolatum for epicutaneous exposures

Induction 2

Concentration / amount:

First induction (intradermally and epicutaenously): Four separate intradermal injections of FCA, emulsified with isotonic saline (to enhance a possible sensitisation) at an area of approx. 2 x 4 cm in the interscapular region, immediately followed by epicutaneous application of the test substance, incorporated in white petrolatum (50 % w/w) or plain white petrolatum (negative control groups) to the sites of the intradermal injections.
Second induction (epicutaenously): 50 % (w/w) in white petrolatum
Challenge exposure (epicutaneously): 50 % (w/w) in white petrolatum

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

petrolatum

Remarks:

white peotrolatum for epicutaneous exposures

Challenge 2

Concentration / amount:

First induction (intradermally and epicutaenously): Four separate intradermal injections of FCA, emulsified with isotonic saline (to enhance a possible sensitisation) at an area of approx. 2 x 4 cm in the interscapular region, immediately followed by epicutaneous application of the test substance, incorporated in white petrolatum (50 % w/w) or plain white petrolatum (negative control groups) to the sites of the intradermal injections.
Second induction (epicutaenously): 50 % (w/w) in white petrolatum
Challenge exposure (epicutaneously): 50 % (w/w) in white petrolatum

No. of animals per dose:

The study performed in two steps. For each step 10 animals were used for as test substance group and another 5 animals as negative control group.
Spare animals: One additional animal per group was kept and administered under the same conditions as the other animals of the respective group. Findings on the spare animals were only to be incorporated into this report if other animals of the test substance group or of the control group would have died spontaneously. Otherwise, the skin reactions of these animals were not used for the interpretation of the results.

Details on study design:

RANGE FINDING TESTS:
To obtain the appropriate concentrations of the test substance for the definitive study, a preliminary test was carried out with 3 female guinea pigs. FCA was administered intradermally and immediately afterwards the test substance was applied epicutaneously (50 % in white petrolatum, w/w) to the sites of the intradermal injections to examine possible systemic toxic effects. Six days later four concentrations of the test substance were administered epicutaneously to the flanks of the animals. The modes of application were the same as in the definitive study. The duration of the epicutaneous exposure was 24 hours. The test substance was applied incorporated in white petrolatum.
48 hours after intradermal injection of FCA and epicutaneous administration of the test substance to the sites of the intradermal injections scores of 3 were given to each of the 3 animals:
Epicutaneous exposure of 50 %, 25 %, 10 % and 5 % formulations of the test substance in white petrolatum (w/w) to the flanks did not elicit any positive skin reaction in any animal 24 adn/or 48 hours after the end of the epicutaneous exposure:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: 24 hours for the first induction (FCA intradermally followed immediately by epicutaneous exposure), 48 hours for the second induction exposure
- Test groups: 1 for each of the two steps
- Control group: 1 for each of the two steps
- Site: area of approx. 2 x 4 cm in the interscapular region
- Concentrations: 50 % (w/w) in white petrolatum, ca. 0.5 g per animal per exposure

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 21
- Exposure period: 24 hours
- Test groups: 1 for each of the two steps
- Control group: 1 for each of the two steps
- Site: left flanks (test substance), right flanks (vehicle)
- Concentrations: 50 % (w/w) in white petrolatum, ca. 0.5 g per animal
- Evaluation: 24 h after the end of the exposure period (Day 23) and a second skin examination further 24 h later (Day 24).

The study was performed in two consecutive steps: In the first step 5 control and 10 test substance animals were used and as after the challenge exposure negative results were obtained, additional 5 control animals and 10 test substance animals were exposed. This procedure is in accordance with the guidelines. The individual animals were allocated to their groups by random numbers. The order of animals for the evaluation of the skin of test and control animals was randomised. A "blind" evaluation was performed.

Test patches (filter papers), 2 cm x 4 cm for the induction exposures or 2 cm x 2 cm for the challenge exposure, with the test substance preparation (test substance group) or with the vehicle (negative control group), were applied. They were fixed with a strip of "Fixomull* stretch" (self adhesive non woven fabric, hypoallergenic, made by Beiersdorf AG, 20245 Hamburg, Germany). On the respective days the hair of the animals was clipped closely on the appropriate application sites with an Aesculap GH 204 electric hair clipper with a 0.1 mm cutter head.

Challenge controls:

For each of the two steps a negative control group (5 animals) was tested in parallel.

Positive control substance(s):

yes

Remarks:

HEXYL CINNAMIC ALDEHYDE (HCA),controls tests performed periodically, data attached to the report

Results and discussion

Positive control results:

Negative control group:
Vehicle site: no positive skin reaction in any animal at any reading time.
Substance site: very sli ght erythema in 1/5 animals 24 hours after the challenge exposure.
Positive control group:
Vehicle site: no positive skin reaction in any animal at any reading time.
Substance site:very slight to severe erythema and/or oedema in 9/10 animals 24 and/or 48 hours after the challenge exposure.
The results of the challenge exposure were decisive for the grading of the potential of sensitisation.
As the rate of animals with positive skin reactions was 20 % in the control group and 90 % in the test substance group, 70 % (90 % - 20 %) of the positive control group animals were regarded as sensitised by "HEXYL CINNAMIC ALDEHYDE".
The results prove the sensitivity of the strain of animals used and the reliability of the experimental technique.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50 % (w/w)

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 % (w/w). No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: none.

Results 2

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

50 % (w/w)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 % (w/w). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Results 3

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50 % (w/w)

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 % (w/w). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

50 % (w/w)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 % (w/w). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

According to the guidelines, the results of this study and to the Directive 2001/59/EC for classification, the test substance "ALUMINIUM TRIFORMATE" needs not to be labelled with "R43 May cause sensitisation by skin contact".

Executive summary:

Aim

The "maximisation test" of B. Magnusson and A. M. Kligman modified according to Maurer & Hess (Th. Maurer and R. Hess (1989) The maximization test for skin sensitization potential-updating the standard protocol and validation of a modified protocol. Fd Chem. Toxic. Vol. 27, No. 12, p. 807-811)

was performed to reveal a possible sensitising potential of "ALUMINIUM TRIFORMATE". The use of this test is justified by the fact, that an LLNA (Local Lymph Node Assay) cannot be performed, as the test substance is poorly soluble in the solvents, commonly used for the LLNA. The same cause is the reason for choosing the modified test (Maurer and Hess) instead of the original GPMT.

Method

Investigations performed were in conformance withtheDirective 96/54/EC, B.6 and with the OECD-Guideline 406.

The study was performed in two consecutive steps. In each step a test substance group with 10 female guinea pigs (Hartley, Crl:Ha) and a negative control group with 5 female guinea pigs (Hartley, Crl:Ha) was used. The administered test substance concentrations were derived from the results of a preliminary test.

Two induction exposures were performed with a one week interval.

First induction exposure: Intracutaneous injection of Freund´s complete adjuvant emulsified with isotonic saline (1:1) and epicutaneous administration of the test substance (50 % in white petrolatum).

Second induction exposure: Preceded by a topical application of 10 % Na-dodecylsulfate in petrolatum for erythema induction 24 h before the epicutaneous administration of the test substance (50 % in white petrolatum).

Control animals were given plain white petrolatum at both inductions.

Challenge exposure: Two weeks after the second induction exposure. Epicutaneous administration of the test substance (50 % in white petrolatum) and the vehicle (white petrolatum), identical for the test substance group and the negative control group.

For the epicutaneous exposures occlusive dressings were used.

 

Results and Conclusion

General

All animals survived till the end of the study. Intradermal injections of Freund's adjuvant caused severe local reactions in all animals, a known effect of the adjuvant. Sensitisation excluded, no other adverse effects were noted.

Skin reactions after the challenge exposure

The results of these skin examinations were decisive for the grading of the potential of sensitisation.

The control sites of all animals of both groups were inconspicuous at each reading time.

In the first step there was a very slight erythema in 1/10 test substance animals (10 %)
24 hours after the challenge exposure. The test substance sites of all other animals in both steps were unaffected at each reading time.

Combined result

1/20 animals of both test substance groups (5 %) was regarded as sensitised.

The test substance "ALUMINIUM TRIFORMATE" was not a sensitiser under the condition of this test.

According to the guidelines, the results of this study and to the Directive 2001/59/EC for classification, the test substance "ALUMINIUM TRIFORMATE" needs not to be labelle dwith "R43 May cause sensitisation by skin contact".