Registration Dossier

Administrative data

Description of key information

The test item is a skin sensitizer in the in vivo test with guinea-pigs.

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

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: Guideline study with GLP.
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
yes
Remarks:
. modification according to Maurer and Hess
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
. modification according to Maurer and Hess
Principles of method if other than guideline:
The "maximisation test" of B. Magnusson and A. M. Kligman modified according to Maurer & Hess was performed to reveal a possible sensitising potential of MPI-ACA. The use of this test is justified by the fact, that a Local Lymph Node Assay LLNA can not 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.
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The "maximisation test" of B. Magnusson and A. M. Kligman modified according to Maurer & Hess was performed to reveal a possible sensitising potential of MPI-ACA. The use of this test is justified by the fact, that a Local Lymph Node Assay LLNA can not 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.
Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Age: Approx. 5 - 7 weeks at the first application
- Weight at the first application: 335 g - 372 g.

- Housing: Group caging in plastic containers (48 cm x 105 cm x 36 cm), partly shaded, 6 (control group) or 11 (test substance group) animals per container.
- Diet: 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: 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: 12 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Mean of 21.9.
- Humidity (%): Mean of 49.5.
- Air changes (per hr): Approx. 12/h
- Photoperiod (hrs dark / hrs light): Only artificial light from 6.00 a.m. to 6.00 p.m.
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
25 % in white petrolatum. The selection of the concentration was based on the results of a preliminary experiment.
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
25 % in white petrolatum. The selection of the concentration was based on the results of a preliminary experiment.
No. of animals per dose:
5 control and 10 test substance animals were used.
1 additional spare animal in each group was kept and treated under the same conditions.
Details on study design:
Day 0: Four separate intradermal injections of FCA, emulsified with isotonic saline (to enhance a possible sensitisation) were given at an area of approx. 2 x 4 cm in the interscapular region. The injections were followed immediately afterwards by an epicutaneous application of the test substance, incorporated in white petrolatum (test substance groups) or plain white petrolatum (negative control groups) to the sites of the intradermal injections. Test patches (filter papers), 2 cm x 4 cm, 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). The treated sites were covered occlusively with a Teflon®-foil and kept in place and fixed with Guinea-Pig Jackets (Hugo Sachs Elektronik- Harvard Apparatus GmbH, 79232, March-Hugstetten, Germany).
24 h afterwards (Day 1) the jackets and the patches were removed.
Effects of the first induction exposure were checked by a skin examination 24 h after the end of the exposure period (Day 2).
On day 6, 24 h prior to the second induction exposure, the exposed skin sites were covered in all animals with a preparation of Na-dodecylsulfate, (E. Merck, 64271 Darmstadt, Germany, item No. 13760; approx. 0.5 g, 10 %, w/w, in white petrolatum), to produce a local hyperaemia.

Second induction exposure, commenced on Day 7:
At the site of the preceding injections of the first induction exposure, an epicutaneous application was performed with the test substance incorporated in white petrolatum (test substance groups) or plain white petrolatum (negative control groups) in analogy to the first induction exposure.
48 h afterwards (Day 9) the jackets and the patches were removed.
Effects of the second induction exposure were checked by a skin examination 24 h after the end of the exposure period (Day 10).

Challenge exposure, commenced on Day 21:
The challenge exposure consisted of two separate epicutaneous applications, identically given to test substance and negative control group animals: One with a test substance preparation to the left flanks and one with the vehicle (white petrolatum) to the right flanks of all animals. Both exposed sites were apart from the exposure sites of the two induction exposures. Test patches (now only 2 cm x 2 cm) and coverings were the same as in both induction exposures.
24 h afterwards (Day 22) the jackets and the patches were removed.
Effects of the challenge exposure were checked by a skin examination 24 h after the end of the exposure period (Day 23) and a second skin examination further 24 h later (Day 24).
Challenge controls:
The challenge exposure consisted of two separate epicutaneous applications, identically given to test substance and negative control group animals.
Positive control substance(s):
yes
Positive control results:
The net rate of animals with a positive skin reaction, regarded as sensitised by hexyl cinnamic aldehyde was 100 % in the last performed test.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25 %
No. with + reactions:
4
Total no. in group:
10
Clinical observations:
no
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 %. No with. + reactions: 4.0. Total no. in groups: 10.0. Clinical observations: no.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25 %
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
no
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: no.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
100 %
No. with + reactions:
10
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
100 %
No. with + reactions:
10
Total no. in group:
10

All animals survived till the end of the study.

No effects of the test substance on body weights was derived from the data.

Immediately after the beginning of all epicutaneous exposures (inductions, challenge) the motor activities of all animals were decreased. This is due to the dressings which restrict the freedom of movement. Soon afterwards the behaviour was regular again. No other abnormal behaviour or clinical signs were detected during the experiment in the animals.

Skin reactions after the first and the second induction exposure

All animals of both groups had severe erythema and oedema in the cranial and caudal rows of the application sites (score "3"). These alterations were attributed to the effects of the adjuvant.

In the middle rows, where the test substance was applied, no skin reactions were noted in any animal at any time.

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
MPI-ACA is a skin sensitizer.
Executive summary:
The "maximisation test" of B. Magnusson and A. M. Kligman modified according to Maurer & Hess was performed to reveal a possible sensitising potential of the test substance. Methods of the EC-directive 96/54, B.6. and the OECD-guideline 406 were applied. A test substance group with 10 female guinea pigs and a negative control group with 5 female guinea pigs were used.

Two induction exposures were performed with a one week interval. Four intradermal injections of FCA emulsified with isotonic saline were given. immediately afterwards the test substance was administered epicutaneously at the injection sites in the intrascapular region. Seven days after completion of the intradermal induction phase, the test substance was administered by topical application to the intrascapular region.

Challenge exposure two weeks after the second induction exposure by epicutaneous administration of the test substance (25 % in white petrolatum) and the vehicle (white petrolatum), identically for the test substance group and the negative control group.

Skin reactions after the challenge exposure: 4/10 animals ( 40 %) of the test substance group were found to have responded positively at the visual examination. In the negative control group no skin reactions were noted. Therefore a total of 40 % of the animals is regarded as sensitised. "MPI-ACA" is regarded as a skin sensitizer under the conditions of the test performed.

Endpoint:
skin sensitisation, other
Remarks:
Experience with the chemical class of the substance and from observations of workers.
Type of information:
other: Experience with the chemical class of the substance and from observations of workers.
Adequacy of study:
other information
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Experience with the chemical class of the substance and from observations of workers.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Experience with the chemical class and from observations of workers.
GLP compliance:
no
Type of study:
other: Experience with the chemical class of the substance and from observations of workers.
Species:
other: Humans
Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.
Executive summary:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)

Respiratory sensitisation

Link to relevant study records
Reference
Endpoint:
respiratory sensitisation, other
Remarks:
Experience with the chemical class of the substance and from observations of workers.
Type of information:
other: Experience with the chemical class of the substance and from observations of workers.
Adequacy of study:
other information
Study period:
2018
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Experience with the chemical class of the substance and from observations of workers.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Experience with the chemical class of the substance and from observations of workers.
GLP compliance:
no
Species:
other: Humans
Interpretation of results:
Category 1 (respiratory sensitising) based on GHS criteria
Conclusions:
Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.
Executive summary:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.

Justification for classification or non-classification

An in vivo skin sensitisation study with guinea-pigs and the experience advises to classify the substance as Skin Sens. 1 and Resp.Sens. 1.