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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
In accordance with Regulation (EU) 2016/1688 preamble (9), adequate information from existing guideline-compliant in vivo skin sensitisation study used to fulfil the standard information requirement point 8.3 Regulation (EC) No 1907/2006.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991-02-05 to 1991-03-14
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 guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted May 12, 1981
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
EEC Directive 84/449 EEC, EEC Publication No. L251, September 1984
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
In accordance with ECHA Guidance R.7a, R.7.3.3 (2017), adequate information from existing guideline-compliant Buehler test used to fulfil the standard information requirement point 8.3 Regulation (EC) No 1907/2006.

Test material

Constituent 1
Reference substance name:
Reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide
EC Number:
915-069-0
Molecular formula:
not applicable
IUPAC Name:
Reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Himalayan albino
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL Ltd., Basel, Switzerland
- Age at study initiation: approximately 11 weeks
- Weight at the end of acclimatisation: 323-413 g
- Housing: Group housing of 2 animals per cage with wire-mesh floors (ITL, Bergen, The Netherlands)
- Diet: Standard guinea pig diet, including ascorbic acid (1600 mg/kg) (LC 23-B, pellet diameter 4mm, Hope Farms, Woerden, The Netherlands) ad libitum. In addition, hay (Broekman Institute, Someren, The Nethrelands) was provided once a week.
- Water: tap-water diluted with decalcified water ad libitum
- Acclimation period: At least five days before start of treatment under test conditions after physical examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 21°C
- Humidity: 55 %
- Air changes (per hr): 15
- Photoperiod: Lighting was 12 hours artificial fluorescent light and 12 hours dark per day.

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Concentration / amount:
25 % test substance in vehicle w/w
Day(s)/duration:
6 hours/ application; 9 applications within 3 weeks
Adequacy of induction:
highest technically applicable concentration used
Challenge
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Concentration / amount:
25 % test substance in vehicle w/w
Day(s)/duration:
6 hours; 10 days after last induction exposure
Adequacy of challenge:
other: highest technically applicable concentration used
No. of animals per dose:
20 in experimental group, 10 in control group
Details on study design:
RANGE FINDING TESTS:

- The objective of this investigation was to identify irritant test substance concentrations suitable for the induction phase of the main study. In addition, a suitable non-irritant concentration of the test substance, by the topical route of administration, was identified for the challenge application. Any systemic toxic effects may also be detected in the primary irritation experiments.
- One animal was treated epicutaneously at the shaved left flank with 0.5 mL of a 50 % concentration of the test substance in propylene glycol using a Metalline patch (Lohman, Neuwied, Germany) mounted on Micropore tape (3M, St. Paul, USA) and held in place with Coban elastic bandage (3M, St. Paul, USA). After 24 hours, the dressings and residual test article were removed using a tissue moistened with tap water. The treated skin was assessed on a numerical basis for erythema and oedema, 24 and 48 hours after bandage removal.
- Four further animals were shaved on the left flank and exposed to 0.05 mL of 50 %, 25 %, 10 % and 5 % (w/w) test substance concentration in propylene glycol, occlusively administered by means of Square chambers (v.d. Bend, Brielle, The Netherlands) mounted on Micropore tape. The bandage was fixed in place by means of Coban elastic bandage. This procedure ensured the intensive contact of the test substance even if it is insoluble in the vehicle used. After 6 hours, the dressing and residual test article were removed using a issue moistened with tap-water. The reaction sites were assessed on a numerical basis for erythema and oedema, 24 and 48 hours after bandage removal.
Immediately after the 24 hour skin reading, the treated areas were re-shaved.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 9
- Exposure period: 6 hours, after each application the remaining test substance was removed from the skin with a tissue moistened with tap water.
- Test groups: one group tested (20 animals)
- Control group: 10 animals, treated as the test group with the omission of the test substance
- Site: scapula region (left side), clipped prior to each application
- Frequency of applications: days 1, 3, 5, 8, 10, 12, 15, 17 and 19
- Duration: 3 weeks
- Concentrations: 25 % (w/w)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: ten days after the last induction exposure (day 29)
- Exposure period: 6 hours. After 6 hours, the dressings and residual test article were removed using a tissue moistened with tap-water.
- Test group: 20 animals
- Control group: 10 animals, treated with 25 % test substance in vehicle and to vehicle alone
- Site: right flank, the test sites were re-shaved with an electric razor after the first reading.
- Concentrations: 25 % (w/w)
- Evaluation (hr after challenge): 24 and 48 hours after removal of the dressings
- Assessment: The challenge sites of experimental and control animals were assessed according to the scale described below for redness and swelling:
-- no skin reaction: 0
-- red spots (scattered reactions): 1
-- moderate but confluent redness: 2
-- redness and swelling: 3
-- intense reddening and swelling: 4

C. OTHER In addition to the skin reactions the following observations and data were recorded:
- Mortality/Viability: once daily
- Body Weights: during acclimatisation and at termination of the study
- Toxicity symptoms: daily

D: INTERPRETATION
- The results evident in test animals at the challenge application(s) were compared with the results evident in control animals.
- A positive skin reaction (grade 1 or more) was considered a sign of sensitisation, provided that such reactions were not observed in the control group.
- The readings after the challenge applications were compared to assess the sensitisation rate i.e. the number of sensitised animals in proportion to the total number of animals of the exposure group.
- The test results were evaluated according to the EEC general classification and labelling requirements for dangerous substances (Annex VI of the EEC Council Directive 67/548/EEC as amended by Directive 83/467/EEC, Sep. 1983).
Challenge controls:
25 % test substance in vehicle and vehicle alone
Positive control substance(s):
yes
Remarks:
Formaldehyde, tested once a year as a sensitivity check of the test system. The most recent test for this study was carried out in April 1990.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25 % (w/w)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25 % (w/w)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25 % (w/w)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25 % (w/w)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.5 % (w/w)
No. with + reactions:
7
Total no. in group:
20

Any other information on results incl. tables

PRIMARY IRRITATION EXPERIMENTS

- No consistent skin reactions. An erythema reaction due to the application of a 50 % (w/w) concentration in the first irritation experiment could not be reproduced in the second irritation experiment.

- No signs of systemic toxicity were observed during the primary irritation experiments. However body weight loss was noted in one of the five animals.

- The choice of propylene glycol as vehicle in this test was based on the fact that propylene glycol produced the best usable suspension.

- The 25 % test article concentration was chosen as the highest usable concentration in the main study, as higher concentrations were too viscous.

MAIN STUDY

- Induction: None of the experimental animals showed skin irritation after the last epicutaneous induction exposure.

- Challenge:

-- Control group: No skin reactions were evident after the challenge exposure.

-- Experimental group: None of the animals showed a positive skin reaction in response to the 25 % concentration.

- Toxicity symptoms/Mortality: No symptoms of systemic toxicity were observed in the animals during the study. No mortality occurred during the study.

- Body weights: The average body weight gain of experimental and control animals was similar.

CONCLUSION

Under the conditions used in this study, the test substance induced no sensitisation.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: Regulation (EC) No 1272/2008 (EU GHS)
Conclusions:
According to the criteria for classification and labelling of Regulation (EC) No 1272/2008, the test substance does not need to be classified as sensitiser.
Executive summary:

In a dermal sensitisation study with substance "reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide" (90 % a.i.) in propylene glycol, young adult female Himalayan albino guinea pigs were tested using the Buehler test method according to OECD Guideline No. 406, adopted May 12 1981 and the EEC Directive 84/449/EEC, Part B.6, September 1984.

None of the animals showed a positive skin reaction in response to the challenge application.

In this study, substance "reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide" (90 % a.i.) is not a dermal sensitizer.

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